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Spinal cord regeneration in the shark

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Title:
Spinal cord regeneration in the shark
Creator:
Gelderd, John Bruce, 1939-
Publication Date:
Language:
English
Physical Description:
x, 79 leaves : ill. ; 29 cm.

Subjects

Subjects / Keywords:
Central nervous system ( jstor )
Dendrites ( jstor )
Goldfish ( jstor )
Lesions ( jstor )
Nerve fibers ( jstor )
Rats ( jstor )
Scars ( jstor )
Sharks ( jstor )
Spinal cord ( jstor )
Sprouting ( jstor )
Dissertations, Academic -- Neuroscience -- UF ( mesh )
Neuroscience thesis Ph.D ( mesh )
Regeneration ( mesh )
Sharks ( mesh )
Spinal Cord ( mesh )
Genre:
bibliography ( marcgt )
non-fiction ( marcgt )

Notes

Thesis:
Thesis (Ph.D.)--University of Florida, 1972.
Bibliography:
Bibliography: leaves 70-78.
General Note:
Typescript.
General Note:
Vita.
Statement of Responsibility:
by John Bruce Gelderd.

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University of Florida
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University of Florida
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Copyright [name of dissertation author]. Permission granted to the University of Florida to digitize, archive and distribute this item for non-profit research and educational purposes. Any reuse of this item in excess of fair use or other copyright exemptions requires permission of the copyright holder.
Resource Identifier:
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25665295 ( OCLC )
AEK6541 ( NOTIS )
AA00004933_00001 ( sobekcm )

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Full Text















SPINAL CORD REGENERATION IN THE SHARK


by
JOHN BRUCE GELDERD











A DISSERTATION PRESENTED TO THE
GRADUATE COUNCIL OF THE
UNIVERSITY OF FLORIDA
IN PARTIAL FULFILLMENT OF THE
REQUIREMENTS FOR THE DEGREE OF
DOCTOR OF PHILOSOPHY









UNIVERSITY OF FLORIDA


1972




SPINAL CORD REGENERATION IN THE SHARK
by
JOHN BRUCE GELDERD
A DISSERTATION PRESENTED TO THE
GRADUATE COUNCIL OF THE
UNIVERSITY OF FLORIDA
IN PARTIAL FULFILLMENT OF THE
REQUIREMENTS FOR THE DEGREE OF
DOCTOR OF PHILOSOPHY
UNIVERSITY OF FLORIDA
1972


DEDICATION
This dissertation is lovingly dedicated to the
memory of my father and to my mother, who so often encouraged
me to "Persevere with your studies son, you shall never
regret it."


ACKNOWLEDGEMENT S
The author takes this opportunity to thank
Dr. J. J. Bernstein for his help and advice in directing
this dissertation.
...the members of his supervisory committee for
their help and moral support.
...Mrs. Gloria Smith, Miss Suzanne Adams and Dr.
Robert Chronister for their expert histological instruction
and assistance.
...Dr. J. I. Thornby for conducting the statistical
analysis.
...Mrs. Linda Burrows for her unselfish help with
the histology, the typing and proofreading of this manuscript
and her constant moral support.
...The Department of Neuroscience and the Center
for Neurobiological Sciences for their financial support.
...and a special thanks to the people of Marineland
of Florida for the use of their facilities, their warm
hospitality and their technical assistance and encouragement.
iii


TABLE OF CONTENTS
Acknowledgements iii
List of Tables v
List of Figures vi
Abstract viii
Introduction 1
Mammalian Studies 2
Inframammalian Studies 12
Elasmobranch Studies 21
Experimental 26
Results 37
Discussion 60
Conclusion 68
Literature Cited 70
Biographical Sketch 79
IV


LIST OF TABLES
Table
1. Summary of Experimental Paradigm 28
2. Summary of degenerating nerve fiber
counts in descending tracts 45
3. Summary of combined left-right bouton
counts on motor horn cell bodies and
primary dendrites 47
4. Summary of strength test results
following spinal cord transection 53
v


LIST OF FIGURES
Figure
1.Location of spinal cord sections removed
for histological analysis 30
2.Apparatus for testing strength of axial
musculature caudal to the site of spinal
cord transection 35
3.Site of spinal cord transection showing
cistern lined with ependymal cells at rostral
stump of cord, small cisterns within the scar
lined with endothelial cells and cisterns
in both stumps of cord, some of which enclose
severed tip of nerve fibers 39
A. 10 Days Postoperative 39
B. 30 Days Postoperative 39
C. 90 Days Postoperative 39
4. A. Large severed nerve fibers showing beading
near their tips and ending in large
spherical globules 42
B. Nerve fibers in rostral stump of spinal
cord beginning their intrusion into the
scar at 20 days postoperative 42
C. Nerve fiber tips growing through the
scar at 30 days postoperative 42
5. Results of bouton counts as revealed
by the Rasmussen stain 49


LIST OF FIGURES
(Continued)
Figure
6.A. Polygraph tracing showing undulatory
movements caudal to transection site 55
B. Polygraph tracing showing response of
axial musculature following stimulation
of barbels of normal animal 55
7. A. Results of strength tests following
spinal cord transection 58
B. Results of timed swimming trials
following spinal cord transection 58
8. A. Comparison between synapse count on
motor horn cell bodies caudal to the
lesion and undulatory strength 67
B. Comparison between synapse count on
motor horn cell dendrites caudal to the
lesion and undulatory strength 67


Abstract of Dissertation Presented to the Graduate
Council of the University of Florida in Partial Fulfillment
of the Requirements for the Degree of Doctor of Philosophy
SPINAL CORD REGENERATION IN THE SHARK
by
John Bruce Gelderd
August, 1972
Chairman: Jerald J. Bernstein, Ph.D.
Major Department: Department of Neuroscience
The shark presents a unique central nervous system
for experimental analysis. The present series of experiments
assess the regenerative capacity of the central nervous
system in the nurse shark (Ginglymostoma cirratum).
The spinal cord was transected at the mid-thoracic
level in 28 nurse sharks. Four animals per group were
sacrificed at intervals of 10, 20, 30, 40, 60 and 90 days
postoperative. Two groups of fish underwent a subsequent spinal
cord retransection at the same site at 90 days postoperative and
were sacrificed 10 and 20 days later. Three sections of
spinal cord were removed from each shark for histological
analysis. A section of spinal cord at the site of lesion
was stained using a modified protargol silver stain to assess
regeneration across the site of lesion. Another spinal cord
section caudal to the lesion site was stained using a
viii


modified Nauta technique to show degenerating descending
nerve fibers. The Rasmussen stain was used for the light
microscopic demonstration of bouton terminaux on motor horn
cells caudal to the lesion. Behaviorally, timed trials for
swimming speed and a strength test for axial musculature
contraction caudal to the lesion site were performed at
five-day postoperative intervals.
Histological analysis showed a neuroglial-pial-
ependymal scar joining the stumps of spinal cord. Regeneration
across the site of lesion did not occur until 40 to 60 days
postoperative. Nerve fibers traversed the lesion site from
both stumps of cord and tended to follow blood vessels and
glial bridges within the scar. The number of descending long
tract nerve fibers reaching an area six spinal segments
caudal to the lesion was small (9-13%) Despite this,
synaptic terminals on motor horn cells caudal to the lesion
showed an increase from 10 to 60 days postoperative (45% of
normal at 10 days postoperative to 92% of normal at 60 days
postoperative).
Immediately upon recovery from anesthesia, all
operated sharks exhibited undulatory movements caudal to the
site of transection while at rest which were independent of
volitional movements rostral to the lesion. These undulatory
IX


movements increased in strength up to 60 days postoperative
at which point they were statistically indistinguishable from
the normal strength of axial musculature. Swimming prowess
was markedly reduced following spinal cord transection and
was never recovered. Undulatory movements were uncontrollable
and proved detrimental to swimming ability. The body caudal
to the site of lesion remained paralyzed in normal attempts
to swim.
Retransection of the spinal cord at 90 days
postoperative showed no change in the strength of axial
musculature caudal to the lesion, timed trials or the number
of boutons on motor horn cells. Comparison between the increase
in undulatory strength and increase in synaptic contacts
on the motor horn cells caudal to the lesion showed a
high correlation (r=.93, PC.01).
It was concluded that the small amount of
regeneration across the site of lesion had no effect on the
swimming behavior of the operated sharks. The increase in
strength of undulatory movements was attributed to the
reestablishment of synaptic contacts on motor horn cells
caudal to the lesion by local, segmental sprouting.
x


INTRODUCTION
Historically, there have been many experiments
and theories aimed at the solution to the perplexing problem
of central nervous system regeneration. One of the earliest
investigators in this area was Ramon y Cajal, who
demonstrated that growth of central nervous system fibers in
embryos resulted from protoplasmic elongation rather than
from fusion of cellular elements (Ramon y Cajal, 1928, 1960).
Ramon y Cajal's conclusions raised the question whether
similar growth would occur following a transection of the
spinal cord of an adult animal. Subsequent studies were
undertaken in both mammalians and inframammalians to assess
anatomical and physiological regeneration of the central
nervous system.
1


MAMMALIAN STUDIES
Following the publications of Ramon y Cajal (1928)
and until the experiments of Sugar and Gerard (1940), little
was added to our knowledge of regeneration in the adult
mammalian spinal cord. During this period, numerous reports
appeared on the regeneration of fetal rat spinal cords.
These experiments met with little or no success and apparent
voluntary function was explained as spinal reflex activity.
Some abortive regeneration was seen, but the regenerated
fibers atrophied before becoming functional (Nicholas and
Hooker, 1928). In 1940, Sugar and Gerard succeeded in
obtaining structural and functional regeneration in the
transected spinal cords of young adult rats. In some of
these animals, muscle and nerve transplants had been oriented
in a longitudinal plane between the severed cord stumps.
These animals were able to sit up, clean their face,
responded by squealing when their tail was pinched and
exhibited coordinated locomotion. Stimulation of the cerebral
peduncles elicited vigorous movements of the hindquarters.
In experiments on spinal cats and dogs (Brown and
2


3
McCouch, 1947), the lack of functional regeneration in the
spinal cord was attributed to the formation of a dense
neuroglial-ependymal scar at the site of lesion. Further
study of this scarring phenomenon was done by Kindle and his
associates (Windle and Chambers, 1950a, 1950b, 1951; Windle,
Clemente and Chambers, 1952; Windle, 1955, 1956; Clemente,
1955). After surgical transection of the spinal cord of
cats and dogs, a dense neuroglial cap or scar formed over
both cut ends of the spinal cord. This scar was composed
of fibroblasts and neuroglia. The relatively avascular
cicatrix between the two stumps of spinal cord was composed
of thick masses of collagenous connective tissue and
completely isolated the cut ends of the cord. It was this
scar which was thought to prevent regeneration and thus
inhibit return of function. Administration of the
bacterially derived polysaccharide, Piromen, prevented the
development of this dense neuroglial scar and permitted the
intrusion of blood vessels into the area of lesion. As a
result of this reduction of the scar, nerve fibers were able
to cross the loose cellular matrix that formed between the
two cut ends of the spinal cord. Freeman (1955) undertook
an extensive series of experiments concerning spinal cord
regeneration in rats. His experiments lasted for more than
15 years. During this time, spinal cord transection was


4
accomplished in over 7,000 rats. Anatomic regeneration and
normal functional return has Occurred in approximately 100
rats. Electrophysiological studies on these rats showed
conduction of impulses in both directions through the site
of lesion. After complete functional recovery (six months
to one year), the "walking paraplegic" rats were injected
with procaine directly into the spinal cord at the site of
transection. The animal again became completely paraplegic.
Retransection of the spinal cord in such rats also returned
them to a paraplegic state. Freeman (1955), Littrell et al.
(1953) and Littrell (1955) transected the spinal cord in
adult cats and contrasted the recovery of non-treated animals
to those treated by intravenous injections of Piromen. Cats
treated with Piromen exhibited return of function beginning
at 2-3 months postoperative and peaked at 9-12 months. After
this point, the animals regressed to typical paraplegic
behavior by 18 months. Histology showed a dense neuroglial-
ependymal scar at the site of transection which was "choking
off" the regenerated fiber tracts. There was never any
return of sensory function. Non-treated animals did not
show any restitution of function or anatomical regeneration.
These studies indicated that the neuroglial-ependymal scar
prevented regeneration in mammals. However, this same
scarring phenomenon also occurred in inframammalian forms that


5
did indeed show both anatomical regeneration and
physiological return of function after spinal cord
transection. It was thought, therefore, that perhaps
mammalian central nervous system neurons lacked sufficient
growth potential to. regenerate lost peripheral processes.
Levi-Montalcini and Brooks (1960) tested the effects of a
protein isolated from the mouse salivary gland upon chick
and mouse sensory ganglia in vitro. Within 12 hours after
injection of this nerve growth factor (NGF) into the medium,
a dense halo of nerve fibers surrounded the explant.
Injection of NGF into intact animals produced hyperplasia
and hypertrophy of sensory and sympathetic neurons. Scott
and Liu (1964) injected NGF and Piromen into kittens after
crushing the dorsal columns. There appeared to be a
definite, direct correlation between the amount and duration
of NGF administration and the regeneration of the sensory
fibers. Anatomical regeneration across the site of lesion
was confirmed electrophysiologically in these kittens
although Scott and Liu did not wait for functional return.
Scott et al. (1966) administered NGF to young rats after dorsal
root crushing and found an increase of 14% in protein production
in dorsal root ganglia as compared to no increase without
administration of NGF. Harvey and Srebnik (1967) found
anatomical and physiological regeneration with return of function


6
following spinal cord compression in rats treated with
L-thyroxine. Non-treated rats showed no regeneration or
return of function. In another rare case of central nervous
system regeneration, Adams et al. (1968, 1969, 1971) cut the
infundibular stalk in ferrets. Degeneration occurred
initially, followed within two weeks by new fibers
regenerating from the hypothalamus through the fibrous scar.
These fibers carried neurosecretory material. During the
one-to three-month postoperative period, fibers grew to a
proximal ectopic infundibular process which formed following
the lesion. In animals kept alive until 12 months
postoperative, the entire neurohypophysis had been
reinnervated and was functional.
If a nerve fiber is severed, there appear to be at
least three distinct reactions: (1) growth does not occur
or it is abortive; (2) the original fiber may regenerate
and reform its synaptic contact or (3) adjacent intact
nerve fibers or cells may develop collateral sprouts and
reinnervate the deafferented tissue (Guth and Windle, 1970).
Liu and Chambers (1958) experimentally demonstrated sprouting
in the spinal cord of the cat following deafferentation
either by adjacent dorsal root section or by corticospinal
tract ablation at cranial levels. They observed that
paraterminal and collateral sprouting was rather generalized


7
in areas which had been deafferented and that the amount
of this sprouting was determined by the extent of the
denervation. Goodman and Horel (1966) showed restricted
sprouting of optic tract projections in the rat after
occipital cortex removal. Schneider (1970) lesioned the
visual cortex or superior colliculus in neonate and adult
hamsters. The neonate hamsters showed sprouting in optic
tract projections and some sparing in visual discrimination
tests in contrast to the adult hamsters which showed little
or no anatomical regeneration or return of function. By
using histochemical fluorescence techniques, Bjorklund and
his colleagues have shown regenerative axon sprouting in
the rat mesencephalon following electrolytic lesions
(Katzman et al., 1971; Bjorklund and Stenevi, 1971) and in
the rat spinal cord following spinal cord compression
(Bjorklund et al., 1971).
Raisman's investigations (1966, 1969a, 1969b)
in the septal nuclei of the adult rat constitute one of the
few ultrastructural studies on collateral sprouting in the
mammalian central nervous system. Afferents from two
separate pathways converge upon the medial septal nucleus.
Fibers originating in the hippocampus pass to the septum
through the medial forebrain bundle. Hippocampal fibers
terminate exclusively on the dendrites of the septal nuclei


-8-
whereas the hypothalamic fibers terminate primarily upon the
cell bodies of the septal nuclei. After lesioning the
hippocampal input, the remaining afferent septal fiber tract
showed a high proportion of axon terminals which made contact
with more than their normal share of postsynaptic units.
This phenomenon was interpreted as a reinnervation by the
remaining hypothalamic input of synaptic sites left open by
lesioning of the hippocampal input. This was corroborated
by lesioning the medial forebrain bundle which produced
degeneration of the remaining synaptic contacts. Raisman
then lesioned the hypothalamic input and found sprouting
of hippocampal fibers in the septum to fill synaptic sites
left by the degenerated hypothalamic input. Moore et al.
(1971) has recently duplicated Raisman's study utilizing
histochemical fluorescence techniques and has corroborated
Raisman's findings. Ultrastructural experiments on rat
spinal cord (Bernstein and Bernstein, 1971) have shown
similar results. Neurons were deafferented by hemisecting
the spinal cord. Shortly after these cells were deafferented,
they began to hypertrophy. In particular, the dendrites
exhibited profuse branching and irregular swelling. Synaptic
spines on the hypertrophied dendrites were also greatly
increased in number. Concomitant with this was the
establishment of large numbers of axodendritic synapses.


9
Furthermore, the axons that terminated near the site of
lesion appeared to arise by way of axonal sprouting from
the region of the spinal cord that was not hemisected. To
ascertain if there was any long tract involvement in the
regenerative process, a group of rats underwent hemisection
of the spinal cord just below T2. Ninety days later, the
spinal cord was hemisected on the same side at C5 and
the spinal cord at vertebral level T2 underwent histological
examination for degeneration. The region immediately rostral
to the T2 lesion showed new, degenerating nerve fibers in
small amounts. The area was also filled with degenerating
axodendritic synapses. These data suggested that the long
tracts were indeed partially involved in the regeneration of
axons to the neurons proximal to the site of the original
lesion. Bernstein stressed, however, that the number of
fibers appeared to be low and that the vast majority of new
synapses originated from segmental sprouting. This study
showed, however, that descending long tract nerve fibers in
the spinal cord of the rat are capable of limited
regeneration to the area immediately rostral to the lesion
site. The regenerating axons in the rat appeared to respond
to nonspecific influences of the hypertrophied dendrites
and established inappropriate connections. The formation
of these synapses then effectively terminated further growth


10
of the axons. Bernstein and Bernstein (in press) have also
shown limited regeneration of axons rostral to the site of
hemisection in the Rhesus monkey spinal cord. Motor horn
cell dendrites immediately rostral to the site of
hemisection showed varicosities. Regenerating axons made
normal as well as aberrant synaptic recombinations with
reactive neurons rostral to the lesion. The most frequent
type of aberrant synaptic complex was a cup-shaped bouton with
a central, large extracellular space between presynaptic and
postsynaptic membranes. In another recent ultrastructural
study, Lund and Lund (1971) found synaptic adjustment in the
superior colliculus following enucleation of neonatal and
adult rats. Little change was observed in the number and
types of synapses in neonatal rats due to synaptogenesis,
but adult rats showed a reinvasion of synaptic sites with
an incomplete return to a normal proportion of synaptic
types.
Although regeneration in the mammalian central
nervous system appears possible, functional return has been
shown in only isolated cases. The majority of experiments
have shown only abortive regeneration. In the instances of
functional return after spinal cord lesion, many are regressive
and the animal returns to a paraplegic state. Sprouting then
appears to be the rule in the mammalian central nervous system,


11
with functional regeneration the rare exception. There is,
as yet, no clear functional significance to the phenomenon
of central nervous system sprouting. McCouch et al. (1955)
has implicated sprouting in spasticity following spinal
cord transection and Schneider (1970) suggests that sparing
of pattern vision behavior in neonatal hamsters is due to
sprouting of visual pathways. However, this evidence is
only suggestive at best and more definitive studies are
required to shed more light on this anatomical phenomenon.


INFRAMAMMALIAN STUDIES
In contrast to the abortive regeneration found in
mammals, inframammalian forms have proven to be a fertile
area for successful central nervous system regeneration
studies. In fact, central nervous system regeneration seems
to be the rule in lower forms with abortive regeneration
the exception. Central nervous system regeneration in bird
embryos has been well documented (Clemente, 1955; Hamburger,
1955) although regeneration in the adult bird occurred only
in the visual system (Cattaneo, 1923). Central nervous
system regeneration studies in reptiles have centered
around the well known phenomenon of tail regeneration in
lizards (Clemente, 1964; Hamburger, 1955). The central
nervous system of amphibia has shown great regenerative
powers. With the noted exception of the adult Anurans, the
amphibia have proven comparable to teleosts in central nervous
system regeneration. After complete transection of the spinal
cord of larval salamanders (Piatt, 1955a, 1955b) or in the
axolotl (Kirsche, 1956), extensive regeneration of fiber
tracts was observed with a concomitant return of normal


13
function. In the adult salamander, regeneration was equally
vigorous (Piatt, 1955a, 1955b). Regeneration of the spinal
cord of the adult newt took place within 30 days with or
without injection of Piromen (Drummond, 1954). Regeneration
in the frog central nervous system has been restricted
to the larval stages (Hooker, 1925) The adult frog has
shown abortive regeneration of the spinal cord after
transection (Piatt and Piatt, 1958; Clemente, 1964).
Regeneration in the frog and toad visual system, however,
has been very specific and successful, both anatomically
and physiologically (Sperry, 1944; Gaze and Jacobson, 1963;
Gaze and Keating, 1969, 1970a, 1970b; Gaze, 1970).
The regenerative capacity of the fish spinal cord
ranks high among the vertebrates. This has been shown
repeatedly through the almost exclusive use of the teleost
as an experimental animal in regeneration studies.
Regeneration in cyclostomes has been largely restricted to
spinal cord regeneration in larval forms (Marn, 1959;
Hibbard, 1963; Niazi, 1963). Spinal cord regeneration in
teleosts has proven so superior to the mammalian nervous
system that what would be considered poor functional or
anatomical recovery in teleosts would undoubtedly be hailed
as strikingly successful in man or any commonly used
laboratory mammal. Regeneration in teleosts does not occur


14
to the same degree in all parts of the central nervous
system. Regeneration throughout the central nervous system
in fish has recently been thoroughly reported by Segaar
(1965) and Bernstein (1970) and will not be repeated here.
The discussion here will be largely restricted to spinal
cord regeneration.
The regenerative capacity of the fish spinal
cord extends from the simple regrowth of axons across the
site of lesion to the complete restitution of neural
cytoarchitectonics, replete with new nerve cells and glia.
Koppanyi and Weiss (1922) carried out spinal cord transections
at a high level in goldfish. The fish were paralyzed caudal
to the lesion for two to three weeks, after which they began
to show signs of return of function. After 60 days, the fish
were behaviorally indistinguishable from normals. Histological
examination showed regeneration of neural pathways which
resulted in the reappearance of normal connections (Koppanyi
and Weiss, 1922; Koppanyi, 1955). Pearcy and Koppanyi (1924)
later cut the entire vertebral column of goldfish with
scissors so that no bony continuity remained between the
regions anterior and posterior to the section. Ten weeks
postoperatively, the fish were again swimming normally.
Hooker (1930, 1932) transected the spinal cord of guppies
less than four days old. He claimed full coordination and


15
integrative movements concomitant with the reestablishment
of nervous connections between the two halves of the body
approximately four days postoperatively. Keil (1940)
transected the spinal cord of adult rainbow fish and claimed
restitution of function beginning from three to twelve days
postoperative with complete restitution of function at
30-40 days postoperative. Ten years later, Kirsche (1950)
confirmed not only the functional but also the morphological
regeneration of the spinal cord in the adult rainbow fish.
Kirsche introduced the method of stimulating the spinal cord
above the site of transection. Tail fin movements were
elicited when the cord, and only the cord was stimulated
above the transection. Those animals which showed no
morphological regeneration also showed no movement of the
caudal fin upon stimulation of the spinal cord above the
transection. Kirsche distinguished various phases during
the course of regeneration. The first phase, which was
apparent approximately four days after the transection,
consisted of a disorganized growth from the severed stumps.
The second phase began about seven days postoperative with
a mitotic increase of the ependymal cells to form "indifferent
neural cells" which in time developed into neuroblasts and
glioblasts. Further differentiation lead to the formation
of normal cells in both proximal and distal stumps. Oriented


16
fibers grew out from these new cells and,approximately 15
days postoperatively, there was evidence for both morphological
continuity and functional recovery (Kirsche, 1950, 1965).
Healey (1962) transected the spinal cord of minnows and noted
the immediate inability of the minnow to change colors upon
background color reversal. Fast color changes were shown to
be under autonomic control and slow color changes were
mediated by hormonal control. Ten days after transection,
the fishes ability to change color increased. After four
months, rapid color changes occurred that were indistinguishable
from normal. Bernstein (1964) has shown a relationship
between age and regenerative capacity of the goldfish spinal
cord. Young goldfish (less than one year old) were able
to reconstitute approximately 90% of the available descending
axons whereas approximately 60% were reconstituted in two-
and three-year-old animals. The ability of the neuroglia
to regenerate and reestablish the diameter of the cord was
also age dependent. The younger goldfish reconstituted the
diameter of the cord almost completely (Bernstein, 1964).
Not only has the spinal cord of the teleost fish regenerated
after being severed, it also has the ability to completely
reconstitute areas of the spinal cord following ablation.
This type of growth pattern has been found in the regeneration
of the caudal neurosecretory system of Tilapia. After removal


17
of the caudal peduncle, tailfin and caudal spinal cord
segments, a new caudal neurosecretory system regenerated.
This system was somewhat abberant but fully functional
(Fridberg et al., 1966).
In a series of experiments on goldfish, it has been
shown that although there was return of function following
spinal cord transection, the morphological regeneration was
less than perfect (Bernstein, 1964; Bernstein and Bernstein,
1968; Bernstein and Gelderd, 1970; Bernstein and Gelderd,
in manuscript). Following spinal cord transection, goldfish
were paralyzed caudal to the site of lesion, descending spinal
tracts degenerated, and synaptic sites on perikaryon and
primary dendrites of motor horn cells 2 cm caudal to the site
of lesion dropped by 50%. Following 60 days regeneration
time, the synaptic complement on motor horn cells was
reestablished although descending fiber tract regeneration
was only 35-50% of normal. A subsequent retransection of the
spinal cord at 60 days postoperative resulted in degeneration
of the new, regenerated descending fibers and concomitant
paralysis caudal to the site of lesion. In contrast, there
was no statistically significant change in the synaptic
complement on motor horn cells 2cm caudal to the site of
lesion. This seemed to indicate that the descending fibers
regenerating into the caudal section of spinal cord did not


18
return to their original synaptic sites on motor horn cell
perikaryon or primary dendrites, but perhaps synapsed instead
on internuncial cells. Return of the normal synaptic complement
on motor horn cell perikaryon or primary dendrites was
relegated to local, segmental sprouting of adjacent fiber
tracts or cells. Those descending fibers which did not
regenerate past the site of lesion appeared to synapse on
cells near the lesion site. When the regenerating axons
reached the site of lesion, they were confronted with
deafferented neurons. The regenerating axons synapsed on
these cells until the synaptic sites were filled. Once the
regenerating axons made these inappropriate synaptic contacts,
they ceased their growth. The mechanism for the cessation
of growth of these axons is thought to be a special case of
contact inhibition (Bernstein and Bernstein, 1968). It must
be stressed again that regeneration in the goldfish central
nervous system is not a uniform phenomenon. Although
transection of the spinal cord was followed by a reduced
regenerative capacity in the number of fibers traversing the
lesion (Bernstein and Gelderd, 1970), lesions of the visual
system were followed by a more specific regenerative process
which initially appeared to be point for point with 100% of
the original optic fibers regenerating (Attardi and Sperry,
1963; Jacobson and Gaze, 1965). Recent studies, however,


19
have shown some deviation in the area of termination of
regenerating optic nerve fibers in the optic tectum of the
goldfish following lesions in the visual system (Yoon, 1971;
Horder, 1971; Sharma, 1972).
In mammalians, the neuroglial-ependymal scar is
thought to be responsible in part for the apparent lack of
regeneration in the central nervous system, forming a dense
barrier between tie cut ends of the spinal cord and thus
preventing regeneration (Brown and McCouch, 1947; Windle,
1955; Windle and Chambers, 1950a, 1950b; Guth and Windle,
1970). Bernstein and Bernstein (1967) investigated the
effect of the neuroglial-ependymal scar on spinal cord
regeneration in goldfish. The spinal cord was completely
transected and a thin teflon disc was placed between the two
cut ends of the spinal cord. The teflon disc remained
between the cut ends for 30 days which was ample time for
regeneration to occur in goldfish. The goldfish were then
operated upon again and the teflon disc removed. These
goldfish were observed for an extended period of time to
determine any return of function caudal to the lesion which
would signify regeneration of the spinal motor tracts. No
regeneration or return of function was observed six months
later. Other goldfish were operated upon again at 30 days
postoperative. The teflon disc was removed and the spinal


20
cord transected one spinal segment rostral to the original
lesion. After 30 days, the spinal cord was observed
histologically to assess regeneration. The descending
spinal motor tracts grew through the second lesion, caudalward
through the isolated- section of the spinal cord and through
the first lesion which was delineated by a substantial
neuroglial-ependymal scar. Hence, the regenerative capacity
of the goldfish spinal cord was not affected by the
neuroglial-ependymal scar acting as a mechanical barrier
(Bernstein and Bernstein, 1967) .


BLASMOBRANCH STUDIES
The shark presents a unique nervous system, both
among the fishes specifically and vertebrates in general.
One trait which is particularly unique to the elasmobranchs
is the reported absence of internuncial cells (Golgi Type II)
in the spinal cord (Kappers et al., 1936; Von Lenhossek,
1892, 1895; Aronson, 1963; Nieuwenhuys, 1964). Presumably,
the vast majority of descending tracts end directly on
motor horn cells without intermediary neurons to intercede or
modulate information from higher centers. Another unique
characteristic restricted to the elasmobranchs is the high
urea content in the blood and sera (Goldstein, 1967; Goldstein
et al., 1968; Rasmussen and Rasmussen, 1967; Smith, 1929;
Rasmussen, 1971). This phenomenon is particularly evident
in marine elasmobranchs. This large concentration of urea
is also reflected in the cerebrospinal fluid (Smith, 1929;
Rasmussen, 1971) and may have a profound effect on the
regenerative capacity of the shark central nervous system
because urea is used to shrink brain tissue during neurological
operations.
21


22
In addition to the above-mentioned characteristics
of the shark central nervous system, this class of fish
shows important advancements in the evolution of the
vertebrate central nervous system. The structure of the
spinal cord in elasmobranchs may be considered a prototype
for that of high vertebrates. Unlike the spinal cord of
cyclostomes which is flat, the spinal cord of elasmobranchs
is round or oval. In addition, these are the first primitive
animals to have myelinated fibers in the spinal cord and
whose dorsal and ventral roots unite outside the vertebral
column to form a mixed root. It is also in this class that
one first finds the division of the gray matter into dorsal
and ventral horns and the first time that all cells of
origin for sensory fibers in the cord lie in extramedullary
spinal ganglia (Kappers, 1906; Kappers et al., 1936; Aronson,
1963; Nieuwenhuys, 1964).
In mammals and other inframammalian forms,
transection of the spinal cord produces paralysis immediately
after transection. In most inframammalians, paralysis is
alleviated by regeneration and return of function after
varying periods of time. In the case of mammals, spinal
walking may occur after weeks or months of recuperation.
The shark, however, has exhibited coordinated undulatory
movements immediately upon recovery from anesthesia after


23
spinal cord transection (Ten Cate and Ten Cate-Kazejawa,
1933; Gray and Sand, 1936a, 1936b; Lissmann, 1946a, 1946b;
Healey, 1957) and was able to swim, using coordinated
movements between those portions of the body rostral and caudal
to the transection. Ten Cate and Ten Cate-Kazejawa (1933)
removed all the muscles in the region of the anterior dorsal
fin of the dogfish (Scyllium cancula and S. catalus) and
transected the spinal cord in the same region. He observed a
locomotory rhythm propogated over the site of the operation,
thereby maintaining coordinated movement between the head and
posterior region of the body. According to Ten Cate and Ten
Cate-Kazejawa (1933), the activity of the posterior region
of the body depended upon tensile stimuli applied to the
posterior musculature whenever an active contraction occurred
in the head region. If this conclusion is justified, then
the normal locomotory rhythm of the dogfish involves the
activity of a chain of peripherally controlled reflexes.
Gray (1936) and Gray and Sand (1936a, 1936b) disagreed with
the hypothesis of Ten Cate and Ten Cate-Kazejawa. They
showed that coordinated responses no longer occurred if two
regions of the body of a dogfish were isolated from one
another by a second spinal cord transection. If both of
these regions were of sufficient length, each exhibited an
independent, spontaneous, automatic activity within the


24
spinal cord. Lissman (1946a, 1946b) showed that locomotory
rhythm could only be abolished in spinal dogfish by a
complete, bilateral rhyzotomy caudal to the transection.
This conclusion appeared to support Ten Cate's hypothesis.
The continued locomotory rhythm after spinal cord transection
in sharks may in part be attributed to its rather low
position on the phylogenetic ladder. Eels have shown an
undulatory behavior after decapitation, having a duration of
only a few seconds (Gray, 1936). Gray has attributed this
phenomenon to injury potentials in the remaining spinal
cord. Typically, spinal eels laid on their side and showed
no coordination between rostral and caudal portions of the
body in normal attempts to swim. Nociceptive stimuli did,
however, cause undulations (Gray, 1936). Little mention has
been made of swimming prowess in the shark with respect to
strength and speed after spinal cord transection. An attempt
to clarify these questions was made during this series of
experiments.
Central nervous system regeneration studies
in elasmobranchs are conspicuous by their absence. Although
lesion studies have been done (Ten Cate and Ten Cate-
Kazejawa, 1933; Gray and Sand, 1936a, 1936b; Lissman,
1946a, 1946b; Healey, 1957; Segaar, 1965; Ebbesson, 1972),
there have been no known experiments concerning


25
shark central nervous system regeneration. Indeed, experimental
work per se on the shark central nervous system is at best
limited. It is this complete lack of central nervous system
regeneration experiments on sharks plus the anatomical and
behavioral uniqueness of their central nervous system that
have prompted me to use them as experimental animals. It was
my intent to use these unique features of the shark central
nervous system to shed some light on the regenerative
process.


EXPERIMENTAL
Materials and Methods
Subjects
Thirty-six male and female nurse sharks
(Ginglymostoma cirratum), approximately two feet in length,
were used. These sharks were trapped in the coastal waters
off Ft. Lauderdale, Florida.
Environment
All fish were kept at Marineland of Florida in an
outside, circular, salt water tank 15 feet in diameter and
six feet in depth. A constantly circulating salt water
system was used to insure proper oxygenation, salinity and
water temperature. Sharks were fed to satiation daily on
cut-up fish.
Operative Procedures
All operated fish were anesthesized with Tricaine
methanesulfonate (MS-222, 1:4000, Finquel, Ayerst Laboratories),
then placed on an operating board. A longitudinal incision
was made at the midline in the midthoracic region at the level
of the trailing edge of the pectoral fin, and the musculature
26


27
dissected away to expose the spinal column. A laminectomy
was performed and the spinal cord transected with a scalpel.
The wound was sutured and powdered sulfathiazole-sulfonilimide
was applied to the suture line to prevent infection. Following
surgery, all animals were also given a 0.1 cc intramuscular
injection of Longicil. Animals were tagged for identification
by attaching a numbered clamp and colored streamers to the
anterior dorsal fin. All sharks, including four normals,
were separated into nine groups (four animals per group)
and were killed by anesthetizing them at 10, 20, 30, 40, 60
and 90 days postoperative and perfusing them with 10% buffered
formalin. Two groups of animals underwent a subsequent
retransection of the spinal cord at the same site at 90 days
postoperative and one group each was killed by the above
fixation method at 10 and 20 days postoperative (Table 1).
Histology
Three sections of the spinal cord were removed
from each shark: a 2 cm section at the site of lesion, a
second section (1 cm in length) six spinal segments caudal
to the site of lesion and a 1 cm section immediately caudal
to the second section (Fig. 1).
The 2 cm section of spinal cord at the site of
lesion was sectioned horizontally at 15q and stained using
a modified Bodian silver technique counterstained with


TABLE 1
SUMMARY OF EXPERIMENTAL PARADIGM
Group
dumber
Number
of
Animals
Spinal
Cord
Transection
Sacrifice Day
Following
Transection
Bodian
Stain
(Nerve Fiber)
Nauta
Stain
(Deqen. Fiber)
Rasmussen
Stain
(Synapses)
Behav. Tests
Five Day
Intervals
3 1
Normal
4
No
1
N/A
X
Normal
X
Normal
X
Normal
2
4
Yes
10
X
X
X
X
3
4
Yes
20
X
X
X
X
4
4
Yes
30
X
X
X
5
4
Yes
40
X
X
X
6
4
Yes
60
X
X
X
7
4
Yes
90
X
X
X
8
4
Yes
10*
X
X
X
X
9
4
Yes
20*
X
X
X
X
TOTAL 36
NOTE: These fish underwent a subsequent spinal cord retransection at the same site at 90 days
postoperative. This number indicates the sacrifice day following the second
2 transection.


Figure 1
- Location of spinal cord sections removed for
histological analysis.


30
Spinal Cord
Section Taken
for Bodian
Site of
Transection
Spinal Cord
Section Taken
for Nauta Stain
Spinal Cord Section
Taken for Rasmussen
Stain
6 spinal segments


31
cresyl-violet and eosin. The regenerative process was
assessed with respect to (1) formation of a neuroglial-
ependymal scar, (2) the effect of the scar on regeneration
and (3) the rate of the regenerative process.
Three descending tracts (tectospinal, thalamospinal
and ventral cerebellospinal tract) were studied in the 1 cm
segment of spinal cord located six spinal segments caudal
to the site of lesion. This section of spinal cord was
histologically analyzed with respect to the number of
degenerating nerve fibers within a given tract following
spinal cord transection (Fig. 1). The spinal cord was
serially sectioned horizontally at 30q and impregnated
using a modified Nauta technique for degenerating nerve
fibers. The degenerative pattern was plotted by drawing a
composite cross-sectional diagram made by examining, in order,
each horizontal section. The histological sections
corresponding to the known anatomical locations of the
given descending tracts were selected and the number of
degenerating fibers counted in each tract at 10 and 20 days
postoperative. These data were compared to the number of
degenerating nerve fibers found 10 and 20 days following
the subsequent retransection (Table 1). The second transection
controlled for local versus long tract input following
regeneration.


32
A quantitative analysis of the synaptic terminals
was done on the perikaryon and primary dendrites of ventral
motor horn cells following spinal cord transection. The
section (1 cm in length) of spinal cord used was taken
immediately caudal to the section used for fiber tract
analysis so that the number of regenerating fibers and the
synaptic profiles could be compared (Fig. 1). The spinal
cord was sectioned coronally at 10q and impregnated using
the Rasmussen stain for the light microscopic demonstration
of bouton terminaux (Rasmussen, 1957) and followed by a
cresyl-violet and eosin counterstain. Counts were made on
only those motor horn cells in which a prominent nucleolus
and primary dendrite could be seen in a given section.
Counts were made on a total of 576 motor horn cells. Sixteen
motor horn cells were counted per shark, utilizing eight
cells on the left side and eight cells on the right side.
Synaptic terminals were counted at 10, 20, 30, 40, 60, and 90
days postoperative and on the two groups of fish retransected
at 90 days postoperative (Table 1). All counts were made on
coded slides to insure unbiased results. The resultant data
were decoded and the levels of significance determined for
intra- and intergroup interactions by using computer
program BMDX63 for multivariate analysis of variance.


33
Behavior
A behavioral analysis of the sharks was done
during the postoperative period including the two groups
retransected at 90 days. The operated sharks were observed
daily while swimming in the tank and compared to normal
sharks with respect to swimming prowess. In addition, two
quantitative tests were performed on all sharks,
preoperatively and at five-day postoperative intervals.
The first test consisted of removing each shark from the
tank and strapping it to a board with that portion of the
body rostral to the lesion firmly held in place. That
portion of the body caudal to the lesion remained unrestrained
with the exception of the caudal peduncle to which a hose
clamp was attached. The clamp was connected by way of a
#10 screw to a Statham load cell assembly (Model UL-4)
which was in turn mounted on a Statham universal force
transducer (Model UC-3). The entire transducer assembly
was securely mounted on the test board. The output of the
force transducer was fed into a two-channel Grass polygraph
recorder. Two electrodes were attached to the paired
barbels located on the underside of the snout of the shark
(Fig. 2). The shark was stimulated using a constant current
stimulator producing a 10 ma pulse of 50 msec duration.
The strength of the response of the caudal body musculature


Figure 2
- Apparatus for testing strength of axial
musculature caudal to the site of spinal
cord transection.


35


36
following stimulation was recorded on the polygraph and
compared to preoperative and normal data. A minimum of
five responses was recorded for each shark on a given
trial day.
The second test consisted of timed swimming
trials. Each shark was placed in the water at one end of a
7' x 3' x 2' tank and the time required for the shark to
swim the length of the tank was recorded. Two consecutive
timed trials were measured on each shark on a given trial
day. Only those trials were counted in which swimming was
uninterrupted over the entire distance. The postoperative
data for both behavioral tests were compared to normal data
in addition to intra- and intergroup interactions among
postoperative groups by utilizing computer program BMD08V
for analysis of variance.


RESULTS
Histological
Site of Lesion
Histological analysis at the site of lesion in
the group of animals sacrificed at 10 days postoperative
showed a dense scar separating the cut ends of the spinal
cord (Fig. 3A). The diameter of the cord in the scar area
was approximately 75% of normal. The scar was composed of
neuroglial, ependymal and pial cells. Blood cells and
phagocytes were also present in abundance throughout the
scar, but no blood vessels were seen within the scar at
this time. Three types of cisterns were found within or
near the site of lesion. A cistern lined with ependymal
cells was visible at the rostral stump of spinal cord.
Several smaller cisterns lined with endothelial cells
were also present throughout the scar. A third type of
space or cistern was found near the lesion site in both
stumps of spinal cord. These spaces were not lined with
cells and some had severed nerve fiber tips within them.
All three types of cisterns were seen in the lesion


Figure 3 Site of spinal cord transection showing cistern
lined with ependymal cells (> at rostral
stump of cord, small cisterns within the scar
(^) lined with endothelial cells and cisterns
in both stumps of spinal cord ( | ) some of which
enclose severed tips of nerve fibers. Bodian
silver stain (X10).
A. 10 days postoperative
B. 30 days postoperative
C. 90 days postoperative cut ventral root
growing from rostral stump of cord into
scar area (f)


39


40
area during the entire postoperative period (Fig. 3A, 3B,
3C). No nerve fibers were found within the scar at this
time. Many large nerve fiber tips were, in fact, found well
back from both cut ends of spinal cord (Fig. 3A). These
large nerve fibers, were beaded at their terminals and ended
in large spherical globules (Fig. 4A). This phenomenon
persisted throughout the postoperative period.
At 20 days postoperative, the scar area resembled
that at ten days. Nerve fibers were seen immediately adjacent
to the scar with some fibers beginning their intrusion into
the scar from both stumps of cord (Fig. 4B).
At 30 days postoperative the cistern lined with
ependymal cells had increased considerably in size and the
diameter of the scar was further reduced (55% of normal).
Nerve fibers were seen penetrating the scar from both
stumps of cord (Fig. 4C) but the center of the scar was
still devoid of nerve fibers.
At 40 days postoperative there was little change
in the appearance of the scar or the density and intrusion
of nerve fibers into the scar area.
There were no discernable differences in the
appearance of the lesion site between 60 and 90 days
postoperative. The scar appeared as a loose cellular matrix
made up of neuroglial and ependymal cells. The ependyma-


Figure 4
A. Large severed nerve fibers showing beading
near their tips and ending in large spherical
globules (|). Bodian silver stain (XI,000).
B. Nerve fibers in rostral stump (RS) of spinal
cord beginning their intrusion into the scar
(S) at 20 days postoperative. Bodian silver
stain (X450) .
C. Nerve fiber tips growing through the scar at
30 days postoperative. Bodian silver stain
(XI, 000) .


42


43
lined cistern at the rostral stump of cord had increased
still more in size (Fig. 3C) but no further constriction
in the diameter of the scar was observed. The increase in
the size of the ependyma-lined cistern in the rostral stump
of spinal cord was probably due to blockage of the flow of
cerebrospinal fluid in the central canal of the spinal cord.
There was a large increase in the number of nerve fibers
within the site of lesion. These nerve fibers were of
small caliber and appeared to completely traverse the scar
by following neuroglial bridges and blood vessels. In those
sharks where the transection was made immediately caudal to
the emergence of a pair of ventral roots, the ventral roots
were severed in the process of transecting the cord.
Severed ventral roots were seen, at 60 and 90 days
postoperative, growing caudally along the edge of the cord
until they reached the lesion site where they grew into the
scar and the caudal stump of spinal cord (Fig. 3C). Nerve
cell bodies were not found within the site of lesion at any
time during the postoperative period.
Nauta Stain
In the process of counting the number of fibers
in the three descending tracts following spinal cord
transection, it was determined that the tectospinal and
thalamospinal tracts could not be effectively counted


44
separately because they were anatomically adjacent to one
another within the spinal cord. Consequently, they were
counted as one tract. There was little evidence of
degeneration 10 days postoperative to both the first
transection and the subsequent retransection. This agreed
with lesion studies in the shark visual system utilizing the
Nauta technique where signs of degeneration following
lesions did not occur until approximately 20 days
postoperative (Ebbesson and Ramsey, 1968; Ebbesson and
Schroeder, 1971) Those sharks sacrificed at 20 days
following both transections showed degeneration within the
three descending tracts and were thus used to make the counts.
Degenerating descending nerve fibers showed typical
irregular beading and droplet formations with concomitant
phagocytic activity typical of lower vertebrates. Ascending
tracts showed no retrograde degeneration.
Nerve fiber counts in the ventral cerebellospinal
tract and the combined tectospinal-thalamospinal tracts are
summarized in Table 2. Degenerating nerve fibers counted
20 days following the first transection represent the normal
complement of axons in each of the respective tracts. Those
nerve fibers counted 20 days following the retransection at
90 days postoperative were the number of regenerated axons
that originated rostral to the lesion site. The number of


45
TABLE 2
SUMMARY OF DEGENERATING NERVE FIBER
COUNTS IN DESCENDING TRACTS
Tract
Numbers
of Fibers
Normal
Regenerated
90 Days
% Regenerated
90 Days
Ventral Cerebellospinal
774
72
9.3
Tectospinal-Thalamospinal
1584
213
13.4


46
degenerating nerve fibers found within the combined
tectospinal-thalamospinal and ventral spinocerebellar tracts
following the second transection were 13.4% and 9.3% respectively
of the number of degenerating nerve fibers found following the
first transection.
Rasmussen Stain
The results of the synaptic terminal counts as
revealed by the Rasmussen stain on cell bodies and primary
dendrites of motor horn cells caudal to the lesion are
summarized in Table 3 and Fig. 5. The average number of
boutons represented was the combined data on both the left
and right sides of the spinal cord since there were no
statistically significant differences (P>.05) between
right and left counts. In addition, there was a high
correlation (r=.974, P<.01) between bouton counts on cell
bodies and primary dendrites.
The number of boutons on cell bodies were
statistically less than normal (£<.05) throughout the
postoperative period, including the retransected groups.
There was, however, a statistically significant increase
in boutons with time after surgery from 10 to 60 days
(P.<.05) but no statistically significant difference occurred
between 60, 90 and the two retransected groups (P^>.05) .
Synaptic terminals dropped to 45% of normal at 10 days


47
TABLE 3
SUMMARY OF COMBINED LEFT-RIGHT BOUTON COUNTS
ON MOTOR HORN CELL BODIES
AND PRIMARY DENDRITES
Days
Postoperative
Combined
Left-Right
Cell Body
Count
(XJ_SEM)
%
of
Normal
Combined
Left-Right
Count Per 10i^
Primary Dendrite
(X+SEM)
10
14.15+0.82
45.06
5.30+0.49
20
17.50^0.47
54.61
5.091,0.98
30
20.50^0.72
65.28
5.92,10.42
40
23.86^0.25
75.98
6.58.+0.17
60
28.96l_0.32
92.22
7.30.+ 0.38
90
29.90^0.12
95.22
7.78^0.37
RETRANSECTION AT
90 DAYS
POSTOPERATIVE
10
29.90i_0.49
95.22
7.421,0.39
20
29.78^0.49
94.84
7.80+0.27
Normal
31.40+0.48
8.51+0.15
%
of
Normal
62.27
59.81
69.56
77.32
85.78
91.42
87.19
91.65


Figure 5 Results of bouton counts as revealed by the
Rasmussen stain. There was a high correlation
(r=.97, P <;01) between counts on motor horn
cell bodies and primary dendrites.


NUMBER OF BOUTONS CELL BODY
49
z
o

z
o
-o
*
>
JO
<
Z
O
DAYS POSTOPERATIVE
NUMBER OF BOUTONS PER 10M


50
postoperative, increased to 92% of normal by 60 days
postoperative and were 95.2% of normal at 90 days
postoperative. The retransected groups sacrificed at 10 and
20 days following retransection were 95.2% and 94.8% of
normal respectively.-
The number of boutons per lOg primary dendrite
following spinal cord transection were significantly less
than normal from 10 to 60 days postoperative (P<.05) and
were significantly greater (P (62.2% of normal at 10 days, 85.7% of normal at 60 days).
The bouton count was 91.4% of normal and statistically
indistinguishable from normal (P>.05) at 90 days
postoperative. Ten days following the retransection,
however, the number of boutons was 87.2% of normal and
statistically less than normal (P<,05). In contrast, there
was no statistically significant difference between normals
and 20 days following the retransection (P>.05).


BEHAVIOR
Daily Observations
Immediately upon recovery from anesthesia, all
operated sharks exhibited undulatory movements caudal to
the site of spinal cord transection. This phenomenon
persisted throughout the postoperative period and was never
observed in normal animals. Although these undulatory
movements occurred caudal to the lesion, there were no
swimming movements caudal to the lesion when the animals
attempted to swim. Forward movement was accomplished by
"walking" along the bottom of the tank using the pectoral
fins or by jerking the body rostral to the lesion left and
right while dragging the caudal portion of the body. Turning
could only be done by walking movements of the pectoral fins.
The undulatory movements during the early
postoperative days (1-30 days) were not strong enough to
move the operated animals. From 30 days postoperative,
however, undulatory movements became strong enough to
propel the sharks forward. The animals did not appear
able to control these undulatory movements. As a result,
51


52
the undulatory movements proved detrimental to their swimming
ability. Stimulation caudal to the lesion site by gentle
prodding or by an inadvertant touch by another shark caused
an increase in undulatory strength which either flipped the
shark over on its back, using the snout as a pivotal point,
or pushed the startled animal into a wall despite its best
efforts to prevent this by using the pectoral fins to
"backpedal" away from the wall. The sharks remained
paralyzed caudal to the transection for the duration of the
postoperative period with respect to normal attempts to swim.
Quantitative Tests
The results of the strength tests and timed swimming
trials are summarized in Table 4.
The strength .tests (Fig. 2) showed two types of
responses. The first response was elicited following
stimulation and was in the form of a single, sharp flexure
of the axial musculature. A second, consecutive stimulation
elicited the same type of response in the opposite direction
(Fig. 6B). This response following stimulation was present
in both normal and operated animals, although much reduced
in the operated animals. This response in operated animals
was probably due to passive conduction of movements caused
by muscle contractions rostral to the lesion. The second
type of response was the previously mentioned undulatory


53
TABLE 4
SUMMARY OF STRENGTH TEST RESULTS
FOLLOWING SPINAL CORD TRANSECTION
UNDULATORY
STRENGTH
DAYS IN KGMS
POSTOPERATIVE (XSEM)
STRENGTH OF
RESPONSE
FOLLOWING
STIMULATION
IN KGMS
(XiSEM)
%
OF
NORMAL
TIMED
SWIMMING
TRIALS
IN SECONDS
(XSEM)
%
OF
NORMAL
1
0.080.01
0.32+0.04
10.63
9.30 0.30
16.12
10
0.210.03
0.290.03
9.63
8.86 0.27
16.70
15
0.21 0.02
0.3210.04
10.63
8.73 0.44
16.95
20
0.26+0.02
0.2710.03
8.97
8.890.38
16.64
25
0.37+0.04
0.3410.04
11.29
9.84+0.47
15.04
30
0.470.06
0.310.02
10.29
8.700.44
17.01
35
0.87 0.12
0.3510.03
11.62
8.060.37
18.36
40
1.14+0.12
0.3810.04
12.63
7.850.30
18.85
50
1.77 0.18
0.4810.05
15.94
7.840.28
18.87
55
1.96+0.24
0.5010.05
16.61
8.430.36
17.55
60
2.66 0.30
0.4910.03
16.27
8.44+0.20
17.53
65
2.13 0.23
0.3410.02
11.29
8.730.31
16.95
70
2.23+0.21
0.3510.02
11.62
8.170.38
18.11
75
2.62 0.63
0.3610.05
11.96
9.17 0.55
16.13
80
2.670.20
0.3510.03
11.62
8.70.32
16.99
85
2.600.18
0.3610.01
11.96
9.08 0.19
16.29
90
2.4610.18
0.3810.02
12.63
9.03 0.09
16.38
RETRANSECTION
AT 90 DAYS POSTOPERATIVE
1
2.2810.16
0.3110.01
10.29
10.45 0.17
14.16
10
2.3210.19
0.3110.02
10.29
9.750.32
15.17
20
2.4410.18
0.3210.01
10.63
9.66 0.26
15.32
Normal
N/A
3.01 0.32
1.48 0.22


Figure 6
A. Polygraph tracing showing undulatory movements
(0.25-0.5 cycles/sec) caudal to transection
site. The top trace indicates when stimulation
occurred. Paper speed = 3mm/sec.
B. Polygraph tracing showing response of axial
musculature following stimulation of the barbels
of a normal animal. The top trace indicates
when stimulation occurred. Paper speed = 3mm/se


55
T
r


56
movements caudal to the site of lesion. These undulatory
movements occurred only in operated animals and were an
almost constant phenomenon requiring no stimulation
(Fig. 6A) .
The resppnse following stimulation was significantly
reduced from normal throughout the postoperative period
(£<;.001) following the first transection and there was no
trend during this postoperative period for any return of
strength caudal to the lesion. There was also no statistically
significant difference (P>.05) between the response following
stimulation at 90 days postoperative and the retransected
groups (Fig. 7A) .
Undulatory movements (0.25-0.5 cycles/sec) were
weak during the early postoperative period, but there was a
statistically significant increase in the strength of
undulatory movements with increased time from 20 to 60 days
postoperative (P_<.05) From 60 days postoperative to 20
days following the retransection, there were no significant
differences between postoperative groups (P>.05). In addition,
the strength attained by undulatory movements at 60 days
postoperative was statistically indistinguishable (P>.05)
from the strength of response following stimulation of
normal animals (Fig. 7A).
A significant increase was observed in the time


Figure 7 -
A. Results of strength tests following spinal
cord transection.
B. Results of timed swimming trials following
spinal cord transection.


58
*
o
<
m
*
m
Z

V
B
STRENGTH IN KGMS OF UNOULATORY


59
required to swim seven feet following spinal cord transection
(P.<.001) Normal animals required an average of 1.48 seconds,
whereas the mean value for transected animals was 8.72
seconds. There was no trend during the postoperative period
for improvement in swimming times (Fig. 7B) and there were
no significant differences between times following the
first and second spinal cord transections (£>.05).


DISCUSSION
These results indicate that following spinal cord
transection, the nurse shark is capable of limited anatomical
regeneration of descending tracts across the site of lesion
(9.3-13.4%) to an area six spinal segments caudal to the
lesion. The time required for nerve fibers to regenerate
across the lesion site was between 40 and 60 days. This was
considerably slower and more incomplete than regeneration in
the teleost spinal cord. Teleosts have shown anatomical
regeneration and return of normal swimming function from
four days postoperative in guppies (Hooker, 1930, 1932)
to approximately 35 days postoperative in goldfish (Bernstein,
1964) although descending fiber tracts in the goldfish
regenerate only 35-49% of the original complement of fibers
within a given tract (Bernstein and Gelderd, 1970).
Limited regeneration did occur in the shark but there was
no return of strength in the axial musculature caudal to
the spinal cord transection following stimulation rostral
to the lesion. The operated sharks were paralyzed caudal
to the lesion in normal attempts to swim. This lack of
60


61
strength caudal to the lesion was also reflected in the
timed swimming trials as there was no trend during the
postoperative period for improvement in swimming times.
In fact, the small amount of anatomical regeneration seen
six spinal segments caudal to the lesion at 90 days
postoperative had little or no effect on strength or
swimming speed as a subsequent retransection at 90 days
postoperative caused no change in these performance parameters
in the ensuing postoperative period.
As was stated previously, only a small number of
nerve fibers regenerated rostro-caudal across the site of
lesion to the area six spinal segments caudal to the lesion
and they did not traverse the lesion site until 40 to 60
days following transection. Despite this, the number of
synapses on motor horn cell bodies and primary dendrites
caudal to the lesion showed an increase beginning at 20 days
postoperative through 60 days postoperative and were highly
correlated (r=.974, P_<.01) .
If synaptic return occurs before the return of
long tract input, what is the source of the increase in the
number of synapses on motor horn cells caudal to the lesion?
The high correlation between primary dendrite and cell body
synaptic counts strongly indicates that both phenomena have
the same origin. The origin for the nerve fibers which


62
replaced lost synaptic contacts must have been caudal to
the site of lesion in the form of local, segmental sprouting.
This hypothesis is further supported by data following the
retransection at 90 days postoperative. There was no
significant change in the number of boutons on motor horn
cell bodies following retransection and a small but statistically
significant (P<.05) drop (91.4% to 87.6% of normal) in boutons
on motor horn cell primary dendrites. This slight drop in
boutons on primary dendrites following retransection was
probably due to degeneration of the small number of regenerated
long tract nerve fibers which synapsed on motor horn cell
dendrites.
If there is no return of swimming prowess and no
return of axial musculature strength following stimulation,
then the question arises as to the functional significance
of this sprouting phenomenon and the return of synaptic
contacts on motor horn cells caudal to the lesion site.
Perhaps the answer to this question lies in the unique
appearance in sharks of undulatory movements caudal to the
lesion following spinal cord transection. Before this
relationship is discussed, however, the anatomical basis
for the undulatory movements will be elaborated.
Unlike other shark studies which claim coordinated
undulatory movements propagated rostro-caudally over the


63
site of lesion (Ten Cate and Ten Cate-Kazejawa, 1933;
Gray, 1936; Gray and Sand, 1936a, 1936b), undulatory movements
in this experiment were observed only caudal to the lesion
site and were independent of body movements rostral to the
lesion. In fact, during the strength tests, the response
following stimulation was often superimposed upon undulatory
movements without affecting the speed or strength of the
undulatory movements.
There are at least two hypotheses relating to the
anatomical basis of undulatory movements in spinal sharks.
Ten Cate and Ten Cate-Kazejawa (1933) claim that the
undulatory movements are propagated over the lesion site
by tensile stimuli applied to posterior musculature when an
active contraction occurs in the head region, implying the
activity of a chain of peripherally controlled reflexes.
Gray (1936) and Gray and Sand (1936a, 1936b) showed that
coordinated responses did not occur if two regions of the
body of a dogfish were isolated from one another by a
second spinal cord transection. Each isolated section of
the body exhibited a spontaneous, independent undulatory
activity. Gray and Sand attributed this to an inherent
undulatory discharge rhythm within the spinal cord.
Lissman (1946a, 1946b) has shown that undulatory movements
caudal to a spinal cord transection in dogfish can only be


64
abolished by a complete bilateral rhyzotomy caudal to the
lesion. The hypothesis of Ten Cate and Ten Cate-Kazejawa
(1933) must be rejected in the present experiment because
undulatory movements persisted without movement rostral to
the lesion. In fact, the undulatory movements were most
prevalent when the shark was at rest on the bottom and were
absent in normal attempts to swim. The two theories need
not be mutually exclusive, however. Lissmann's studies
showed that dorsal root input is mandatory for the maintenance
of undulatory movements. Thus it appears from the present
data that the spinal cord of the shark has an inherent
undulatory discharge pattern modulated by local sensory
input and input from brain centers. If the major effect
of the brain on this undulatory discharge pattern is
inhibitory, transecting the spinal cord will release the
caudal section of spinal cord from these inhibitory influences,
thus allowing the inherent discharge pattern to be exhibited
in the form of undulatory movements caudal to the lesion site.
If the synaptic sites left vacant by spinal cord transection
are replaced with excitatory synapses from dorsal root
fibers or indigenous spinal tracts by way of sprouting, an
increase in the discharge pattern should occur with a
resultant increase in the strength of undulatory movements.
Undulatory movements in the present experiment began


65
immediately upon recovery from anesthesia and increased
in strength up to 60 days postoperative after which they
leveled off. If the postoperative increase in the strength
of undulatory movements is compared to the postoperative
increase in synaptic complement on motor horri cells caudal
to the lesion and the resultant data plotted on a graph
(Fig. 8A, 8B), the curves of the two postoperative phenomena
are highly correlated. Comparison between boutons on cell
bodies and undulatory movements has a correlation coefficient
of r=.930 (P_<. 01) and comparison between boutons on motor
horn cells primary dendrites and undulatory movements results
in a correlation coefficient of r=.91 (:P<:01) .
It is therefore highly probable from these data
that the synaptic return on motor horn cells by way of
local sprouting is responsible for the increase in the
strength of undulatory activity caudal to the site of lesion.
To further support this hypothesis, retransection at 90 days
had a minimal effect on both synaptic complement and
undulatory movements (Fig. 8A, 8B).


Figure 8 -
A. Comparison between synapse count on motor
horn cell bodies caudal to the lesion and
undulatory strength (r=.93, P_<.01) .
B. Comparison between synapse count on motor
horn cell dendrites caudal to the lesion
and undulatory strength (r=.91, P_<. 01).


NUMBER OF BOUTONS PER 10i<
67
>-
Q
o
z
O
-
D
O
0D
£
3
Z
r 3.0
-2.4
18
-1.2
-0.6
DAYS POSTOPERATIVE
QC
Q
Z
Q
>
oc
<
*
ac
Q.
u.
o
z

o
z
r 3-0
-2.4
1.8
-1.2
-0.6
DAYS POSTOPERATIVE
B
STRENGTH IN KGMS OF UNDULATORY STRENGTH IN KGMS OF UNDULATORY
MOVEMENTS ^ MOVEMENTS


CONCLUSION
Regeneration in the shark spinal cord following
spinal cord transection appears to lie somewhere between
the abortive regeneration usually seen in mammals and the
vigorous regeneration and return of function typical of
teleosts. Although anatomical regeneration of nerve
fibers across the site of lesion does occur, the functional
ramifications are negligible and for all practical purposes
the shark remains paralyzed caudal to the lesion when
attempting to swim. The poor anatomical regeneration and
lack of functional return in sharks is surprising. A
general rule of thumb which is well documented is that
the lower on the phylogenetic scale, the more vigorous
and complete is the central nervous system regenerative
process. The reasoning for this is that more primitive animals
reportedly possess more undifferentiated, pleuri-potential
cells capable of differentiation into neural elements. The
elasmobranchs occupy the third rung up on the vertebrate
phylogenetic ladder immediately below the teleosts, yet show
68


69
a postoperative recovery following spinal cord transection
more akin to mammals than fish.
Perhaps the most significant and interesting
result of this experiment is the strong indication of a
functional correlation between the return of synapses on
motor horn cells caudal to the lesion and the increase in
undulatory strength during the postoperative period.
Although functional correlates have been suggested for the
phenomenon of sprouting in the mammalian central nervous
system (McCouch et al., 1955; Schneider, 1970) the evidence
presented in this experiment is perhaps the most conclusive
to date of a functional correlate to sprouting in the
vertebrate central nervous system.


LITERATURE CITED
Adams, J. H., P. M. Daniel and M. M. L. Prichard 1968
Regrowth of nerve fibers in the neurohypophysis:
Regeneration of a tract of the central nervous system.
J. Physiol. (London), 198:4P-5P.
Adams, J. H., P. M. Daniel and M. M. L. Prichard 1969
Degeneration and regeneration of hypothalamic nerve
fibers in the neurohypophysis after pituitary stalk
section in the ferret. J. Comp. Neurol., 135:121-144.
Adams, J. H., P. M. Daniel and M. M. L. Prichard 1971
Changes in the hypothalamus associated with regeneration
of the hypothalamo-neurohypophysial tract after
pituitary stalk section in the ferret. J. Comp. Neurol.,
142:109-124.
Aronson, L. R. 1963 The central nervous system of sharks
and bony fishes with special reference to sensory and
integrative mechanisms. In Sharks and Survival (Ed.
Perry W. Gilbert) pp. 165-241, D. C. Heath and Co.,
Boston, Massachusetts.
Attardi, D. G. and R. Sperry 1963 Preferential selection
of central pathways by regenerating optic fibers.
Exp. Neurol., 7:46-64.
Bernstein, J. J. 1964 Relation of spinal cord regeneration
to age in adult goldfish. Exp. Neurol., 9:161-174.
Bernstein, J. J. 1970 Anatomy and Physiology of the central
nervous system. In Fish Physiology Vol. IV (Ed. W. S.
Hoar and D. J. Randall) pp. 1-90, Academic Press,
New York, London.
Bernstein, J. J. and M. E. Bernstein 1967 Affect of the
glial ependymal scar and teflon arrest on the
regenerative capacity of the goldfish spinal cord.
Exp. Neurol., 19:25-32.


71
Bernstein, J. J. and M. E. Bernstein 1968 Contact inhibition:
A mechanism of abortive regeneration in the goldfish
spinal cord. Anat. Rec. 160:315-316.
Bernstein, J. J. and M. E. Bernstein 1971 Axonal regeneration
and formation of synapses proximal to the site of lesion
following hemisection of the rat spinal cord. Exp.
Neurol., 30:336-351.
Bernstein, M. E. and J. J. Bernstein 1972 Regeneration in
the spinal cord of the monkey. (In press).
Bernstein, J. J. and J. B. Gelderd 1970 Regenerative
capacity of long spinal tracts in the goldfish. Brain
Res., 19:21-26.
Bernstein, J. J. and J. B. Gelderd 1972 Synaptic complement
formation following regeneration of the goldfish spinal
cord. (In manuscript).
Bjorklund, A. R. Katzman, U. Stenevi and K. A. West 1971
Development and growth of axonal sprouts from noradrenaline
and 5-hydroxytryptamine neurones in the cat spinal cord.
Brain Res., 31:21-33.
Bjorklund, A. and U. Stenevi 1971 Growth of central
catecholamine neurones into smooth muscle grafts in
the rat mesencephalon. Brain Res., 31:1-20.
Brown, J. 0. and G. P. McCouch 1947 Abortive regeneration
of the transected spinal cord. J. Comp. Neurol., 87:131-137.
Cattaneo, D. 1923 I fenomeni degenerativi nelle vie visive
in seguito a lesioni del ervo ottico. Riv. Pat. Nerv.
28:61-118.
Clemente, C. D. 1955 Structural regeneration in the mammalian
central nervous system and the role of neuroglia and
connective tissue. In Regeneration in the Central Nervous
System (Ed. William F. Windle) pp. 147-161, Charles C.
Thomas, Springfield, Illinois.
Clemente, C. D. 1964 Regeneration in the vertebrate central
nervous system. Intern. Rev. Neurobiol., 6:257-301.
Drummond, C. D., Jr. 1954 The influence of piromen on the
regeneration of the spinal cord in adult Triturus
viridescens. Undergrad. Hon. Thesis, Brown Univ.,
' -T.. 'I.


72
Ebbesson, Sven 0. E. 1972 New insights into the organization
of the shark brain. Comp. Biochem. and Physiol. 42:121-130.
Ebbesson, Sven O. E. and John S. Ramsey 1968 The optic
tracts of two species of sharks (Galeocerdo cuvier and
Ginglymostoma cirratum). Brain Res. 8:36-53.
Ebbesson, Sven 0. E. and Dolores M. Schroeder 1971
Connections of the nurse shark's telencephalon. Science
173:254-256.
Freeman, L. W. 1955 Functional recovery in spinal rats.
In Regeneration in the Central Nervous System (Ed.
William F. Windle) pp. 195-207, Charles C. Thomas,
Springfield, Illinois.
Fridberg, G., R. S. Nishioka and W. R. Fleming 1966
Regeneration of the caudal neurosecretory system in
the cichlid teleost Tilapia mossambica. J. Exp.
Zool. 162:311-336.
Gaze, R. M. 1970 The Formation of Nerve Connections.
Academic Press, London and New York.
Gaze, R. M. and M. Jacobson 1963 The path from the retina
to the ipsilateral optic tectum of the frog. J.
Physiol. 165:73-74.
Gaze, R. M. and M. J. Keating 1969 The depth distribution
of visual units in the tectum of the frog following
regeneration of the optic nerve. J. Physiol. 200:128-129.
Gaze, R. M. and M. J. Keating 1970a Further studies on the
restoration of the contralateral retinotectal projection
following regeneration of the optic nerve in the frog.
Brain Res., (In Press).
Gaze, R. M. and M. J. Keating 1970b Regenerated visual
units in the frog. Brain Res., (In Press).
Goldstein, L. 1967 Urea Biosynthesis in Elasmobranchs.
In Sharks, Skates and Rays (Ed. Perry W. Gilbert,
Robert F. Mathewson and David P. Rail) John Hopkins
Press, Baltimore, Maryland.
Goldstein, L., W. W. Oppelt and T. H. Maren 1968 Osmotic
regulation and urea metabolism in the lemon shark
Neqaprion brevirostris. Am. J. Physiol. 215:1493,1497.


73
Goodman, D. C. and J. A. Horel 1966 Sprouting of optic
tract projections in the brain stem of the rat.
J. Comp. Neurol., 127:71-78.
Gray, J. 1936 Studies in animal locomotion. IV. The
neuromuscular mechanism of swimming in the eel.
J. Exp. Biol., 13:170.
Gray, J. and A. Sand 1936a The locomotory rhythm of the
dogfish (Scyllium canicula). J. Exp. Biol. 13:200-209.
Gray, J. and A. Sand 1936b Spinal reflexes of the dogfish
(Scyllium canicula). J. Exp. Biol. 13:210-217.
Guth, L. and W. F. Windle 1970 The enigma of central nervous
regeneration. Exp. Neurol. 28:1-43, Supp. 5.
Hamburger, V. 1955 Regeneration in the central nervous
system of reptiles and of birds. In Regeneration in
the Central Nervous System (Ed. William F. Windle)
pp. 47-53, Charles C. Thomas, Springfield, Illinois.
Harvey, J. E. and H. H. Srebnik 1967 Locomotor activity
and axon regeneration following spinal cord compression
in rats treated with L-thyroxine. J. Neuropath. Exp.
Neurology 26:661-668.
Healey, E. G. 1957 The Nervous System. In The Physiology
Of Fish Vol. 2 (Ed. Margaret E. Brown) pp. 1-119,
Academic Press Inc., New York.
Healey, E. G. 1962 Experimental evidence for regeneration
following spinal section in the minnow (Phoxinus phoxinus).
Nature 194:395-396.
Hibbard, E. 1963 Regeneration of the severed spinal cord
of chordate larvae of Petromyzon marinus. Exp. Neurol.,
7:175-185.
Hooker, D. 1925 Studies on regeneration in the spinal cord.
III. Reestablishment of anatomical and physiological
continuity after transection in frog tadpoles. J.
Comp. Neurol. 38:315-347.
Hooker, D. 1930 Physiological reactions of goldfish with
severed spinal cord. Proc. Soc. Exp. Biol. Med.,
28:89-90.


74
Hooker, D. 1932 Spinal cord regeneration in the young
rainbow fish Lebistes Reticulatus. J. Comp. Neurol.,
56:277-295.
Horder, T. J. 1971 Retention by fish optic nerve fibers
regenerating to new terminal sites in the tectum of
"chemospecific" affinity for their original sites.
J. Physiol. (London) 216:53-55P.
Jacobson, M. and R. M. Gaze 1965 Selection of appropriate
tectal connections by regenerating optic nerve fibers
in adult goldfish. Exp. Neurol., 13:418-430.
Kappers, Ariens C. U. 1906 The structure of the teleostian
and selachian Brain. J. Comp. Neurol, and Psychology.
16:1-110.
Kappers, Ariens C. U., G. C. Huber and E. C. Crosby 1936
The Comparative Anatomy of the Nervous System of
Vertebrates Including Man. Vol. 1, The Macmillan Co.,
New York.
Katzman, R., A. Bjorklund, Ch. Owman, U. Stenevi and K. A.
West 1971 Evidence for regenerative axon sprouting
of central catecholamine neurons in the rat mesencephalon
following electrolytic lesions. Brain Res., 25:579-596.
Keil, J. H. 1940 Functional spinal cord regeneration in
adult Rainbow fish. Soc. Exp. Biol. Med., 43:175-177.
Kirsche, W. 1950 Die regenerativen Vorgange am Ruckenmark
eruachsever Teleostier nach operativer Kontinuitat.
Strennung. Z. Mikroskop. Anat. Forsch. 56:190-265.
Kirsche, W. 1956 Experimentelle Untersuchunger uber die
regeneration des durchtrennten Ruckenmarkes von
Amblystoma mexacanum. Z. Mikroskop. Anat. Forsch.
65:512-586.
Kirsche, W. 1965 Regenerative Vorgange im Gehirn und
Ruckenmark. Ergeb. Anat. Entiv-gesch. 38:143-194.
Koppanyi, T. 1955 Regeneration in the central nervous
system of fishes. In Regeneration in the Central
Nervous System (Ed. William F. Windle) pp.3-19,
Charles C. Thomas, Springfield, Illinois.


75
Koppanyi, T. and P. Weiss 1922 Funktionelle regeneration
des Ruckenmarkes bei anamniern. Anz. Akad. Wiss. Wien.
Math. Naturre Kl. 59:206-219.
Levi-Montalcini, R. and B. Brooks 1960 Excessive growth
of the sympathetic ganglis evoked by a protein isolated
from the mouse salivary gland. Proc. Nat. Acad. Sci.
(Wash.) 46:373-384.
Lissmann, H. W. 1946a The neurological basis of the
locomotory rhythm in the spinal dogfish (Scylliuro
cancula, Acanthius vulgaris) I. Reflex Behavior.
J. Exp. Biol. 23:143-161.
Lissmann, H. W. 1946b The neurological basis of the
locomotory rhythm in the spinal dogfish (Scyllium
cancula, Acanthius vulgaris) II. The effect of
deafferentation. J. Exp. Biol. 23:162-176.
Littrell, J. L. 1955 Apparent functional restitution in
piromen treated spinal cats In Regeneration in the
Central Nervous System (Ed. William F. Windle)
pp. 219-228, Charles C. Thomas, Springfield, Illinois.
Littrell, J. L., D. Bunnell, W. F. Agnew, J. O. Smart and
W. F Windle 1953 Effects of a bacterial pyrogen on
hind-limb function in spinal cats. Anat. Rec., 115:430-436.
Liu, C. N. and W. W. Chambers 1958 Intraspinal sprouting of
dorsal root axons. Arch. Neurol. 79:46-61.
Lund, R. D. and J. S. Lund 1971 Synaptic adjustment after
deafferentation of the superior colliculus of the rat.
Science, 171:804-807.
Marn, K. 1959 Regeneration capacity of the spinal cord
in Lampetra fluviatilis larvae. Folia Biol., 7:179-189.
McCouch, G. P., G. M. Austin and C. Y. Liu 1955 Sprouting
of new terminals as a cause of spasticity. Am. J.
Physiol. 183:642.
Moore, R. Y., A. Bjorklund and U. Stenevi 1971 Plastic
changes in the adrenergic innervation of the rat
septal area in response to denervation. Brain Res.
(In Press).


76
Niazi, I. A. 1963 The histology of tail regeneration in
the ammocoetes. Can. J. Zool., 41:125-145.
Nicholas, J. S. and D. Hooker 1928 Progressive cord
degeneration and collateral transmission of spinal
impulses following section of the cord in albino
rat fetuses. Anat. Rec. 38:24-32.
Nieuwenhuys, R. 1964 Comparative anatomy of the spinal
cord. Prog. Brain Res. 11:1-57.
Pearcy, J. F. and T. Koppanyi 1924 A further note on
regeneration of the cut spinal cord in fish. Proc.
Soc. Exp. Biol. Med., 22:17-19.
Piatt, J. 1955a Regeneration in the central nervous
system of amphibia. In Regeneration in the Central
Nervous System (Ed. William F. Windle) pp. 20-46,
Charles C. Thomas, Springfield, Illinois.
Piatt, J. 1955b Regeneration of the spinal cord in the
salamander. J. Exp. Zool., 129:177-207.
Piatt, J. and M. Piatt 1958 Transection of the spinal
cord in the adult frog. Anat. Rec. 131:81-95.
Raisman, G. 1966 The connexions of the septum. Brain
89:317-348.
Raisman, G. 1969a A comparison of the mode of termination
of the hippocampal and hypothalamic afferents to the
septal nuclei as revealed by the electron microscopy
of degeneration. Exp. Brain Res. 7:317-343.
Raisman, G. 1969b Neuronal plasticity of the septal nuclei
of the adult rat. Brain Res. 14:25-48.
Ramon y Cajal, S. 1928 Degeneration and Regeneration of the
Nervous System. 1:47-51.
Ramon y Cajal, S. 1960 Studies on Vertebrate Neurogenesis,
L. Guth, Translator, Charles C. Thomas, Springfield,
Illinois.


77
Rasmussen, G. L. 1957 Selective silver impregnation of
synaptic endings In New Research Techniques of
Neuroanatomy, (Ed. W. F. Windle) pp. 27-39, Charles
C. Thomas, Springfield, Illinois.
Rasmussen, L. E. 1971 Organ distribution of exogenous
^C-urea in elasmobranchs with special regard to the
nervous system. Comp. Biochem. Physiol. 40A:145-154.
Rasmussen. L. E. and R. A. Rasmussen 1967 Comparative
protein and enzyme profiles of the cerebrospinal fluid,
extradural fluid, nervous tissue and sera of
elasmobranchs In Sharks, Skates and Rays (Ed. Perry W.
Gilbert, Robert F. Mathewson and David P. Rail)
pp. 361-380, John Hopkins Press, Baltimore, Maryland.
Schneider, G. E. 1970 Mechanisms of functional recovery
following lesions of visual cortex and superior
colliculus in neonate and adult hamsters. Brain,
Behavior and Evol. 3:295-323.
Scott, D., Jr., E. Gutmann and P. Horsky 1966 Regeneration
in spinal neurons: Proteosynthesis following nerve
growth factor administration. Science 152:787-788.
Scott, D. and C. N. Liu 1964 Factors promoting regeneration
of spinal neurons: Positive influence of nerve growth
factor. Prog, in Brain Res. 13:127-150.
Segaar, J. 1965 Behavioral aspects of degeneration and
regeneration in fish brain: A comparison with higher
vertebrates. Prog. Brain Res. 14:143-231.
Sharma, S. C. 1972 Reformation of retinotectal projections
after various tectal ablations in goldfish. Exp.
Neurol. 34:171-182.
Smith, H. 1929 The composition of the body fluids in
elasmobranchs. J. Biol. Chem. 81:407-419.
Sperry, R. W. 1944 Optic nerve regeneration with return
of vision in anurans. J. Neurophysiol. 7:57-70.
Sugar, 0. and R. W. Gerard 1940 Spinal cord regeneration
in the rat. J. Neurophysiol. 3:1-19.


78
Ten Cate, J. and Ten Cate-Kazejawa 1933 La coordination des
mouvements locomoteurs apres la section transversale de
la moelle epiniere chez les requins. Arch. Neerl.
Physiol. 18:15-23.
Von Lenhossek, M. 1892 Beobachtungen an den spinalganglien
und deni Ruchenmark von Pristiurusembryonen. Anat.
Anz. Bd. 7:519.
Von Lenhossek, M. 1895 Der fienere bau des nervensystems
im lechte neuester Forschungen. Aufl. Fischer, Berlin.
Windle, W. F. 1955 Comments on regeneration in the human
central nervous system. In Regeneration in the Central
Nervous System (Ed. William F. Windle) pp. 265-272,
Charles C. Thomas, Springfield, Illinois.
Windle, W. F. 1956 Regeneration of axons in the vertebrate
central nervous system. Physiol. Rev. 36:427-440.
Windle, W. F. and W. W. Chambers 1950a Regeneration in the
spinal cord of the cat and dog. J. Comp. Neurol.
93:241-257.
Windle, W. F. and W. W. Chambers 1950b Spinal cord
regeneration associated with a cellular reaction induced
by administration of a purified bacterial pyrogen.
Abst. V. Internat. Anat. Comg., Oxford, p. 196.
Windle, W. F. and W. W. Chambers 1951 Regeneration in the
spinal cord of the cat and dog. Arch. Neurol. Psychiat.,
Chi., 65:261-262.
Windle, W. F., C. D. Clemente and W. W. Chambers 1952
Inhibition of formation of a glial barrier as a means
of permitting peripheral nerve to grow into the brain.
J. Comp. Neurol. 96:359-370.
Yoon, M. 1971 Reorganization of retinotectal projection
following surgical operations on the optic tectum in
goldfish. Exp. Neurol. 33:395-411.


BIOGRAPHICAL SKETCH
John Bruce.Gelderd was born during a German
bombing raid in Wasquehal, France on September 21, 1939.
He attended grade school in Broadview, Illinois, and Clearwater,
Florida, and graduated from Clearwater High School in 1957.
After a 4^ year stint in the U. S. Air Force, he attended
St. Petersburg Jr. College from September, 1962, to May,
1964. He entered the University of Florida in September,
1964 and received a bachelor's degree in zoology in 1967.
He began his graduate studies toward the Doctor of
Philosophy degree at the University of Florida in September,
1968.
He has accepted a faculty position in the
Department of Anatomy at Louisiana State University in
New Orleans, Louisiana.
79


I certify that I have read this study and that in my
opinion it conforms to acceptable standards of scholarly
presentation and is fully adequate, in scope and quality,
as a dissertation for the degree of Doctor of Philosophy.
/cv/V-yxvi
J/'J. Bernstein, Chairman
Asbciate Professor of Neuroscience
I certify that I have read this study and that in my
opinion it conforms to acceptable standards of scholarly
presentation and is fully adequate, in scope and quality,
as a dissertation for the degree of Doctor of Philosophy.
A
F. A. King, Chairman of
Department of Neuroscience
I certify that I have read this study and that in my
opinion it conforms to acceptable standards of scholarly
presentation and is fully adequate, in scope and quality
as a dissertation for the degree of Doctor of Philosophy.
C./JJ. Vierck,
Associate Processor of Neuroscience
I certify that I have read this study and that in my
opinion it conforms to acceptable standards of scholarly
presentation and is fully adequate, in scope and quality,
as a dissertation for the degree of Doctor of.Philosophy.
/ S- /CiUy^t^
John B. Munson
Assistant Professor, Neuroscience


This dissertation was submitted to the Department of
Neuroscience in the College of Medicine and to the Graduate
Council, and was accepted as partial fulfillment of the
Dean, Graduate School


Full Text
SPINAL CORD REGENERATION IN THE SHARK
by
JOHN BRUCE GELDERD
A DISSERTATION PRESENTED TO THE
GRADUATE COUNCIL OF THE
UNIVERSITY OF FLORIDA
IN PARTIAL FULFILLMENT OF THE
REQUIREMENTS FOR THE DEGREE OF
DOCTOR OF PHILOSOPHY
UNIVERSITY OF FLORIDA
1972

DEDICATION
This dissertation is lovingly dedicated to the
memory of my father and to my mother, who so often encouraged
me to "Persevere with your studies son, you shall never
regret it."

ACKNOWLEDGEMENT S
The author takes this opportunity to thank
Dr. J. J. Bernstein for his help and advice in directing
this dissertation.
...the members of his supervisory committee for
their help and moral support.
...Mrs. Gloria Smith, Miss Suzanne Adams and Dr.
Robert Chronister for their expert histological instruction
and assistance.
...Dr. J. I. Thornby for conducting the statistical
analysis.
...Mrs. Linda Burrows for her unselfish help with
the histology, the typing and proofreading of this manuscript
and her constant moral support.
...The Department of Neuroscience and the Center
for Neurobiological Sciences for their financial support.
...and a special thanks to the people of Marineland
of Florida for the use of their facilities, their warm
hospitality and their technical assistance and encouragement.
iii

TABLE OF CONTENTS
Acknowledgements iii
List of Tables v
List of Figures vi
Abstract viii
Introduction 1
Mammalian Studies 2
Inframammalian Studies 12
Elasmobranch Studies 21
Experimental 26
Results 37
Discussion 60
Conclusion 68
Literature Cited 70
Biographical Sketch 79
IV

LIST OF TABLES
Table
1. Summary of Experimental Paradigm 28
2. Summary of degenerating nerve fiber
counts in descending tracts 45
3. Summary of combined left-right bouton
counts on motor horn cell bodies and
primary dendrites 47
4. Summary of strength test results
following spinal cord transection 53
v

LIST OF FIGURES
Figure
1.Location of spinal cord sections removed
for histological analysis 30
2.Apparatus for testing strength of axial
musculature caudal to the site of spinal
cord transection 35
3.Site of spinal cord transection showing
cistern lined with ependymal cells at rostral
stump of cord, small cisterns within the scar
lined with endothelial cells and cisterns
in both stumps of cord, some of which enclose
severed tip of nerve fibers 39
A. 10 Days Postoperative 39
B. 30 Days Postoperative 39
C. 90 Days Postoperative 39
4. A. Large severed nerve fibers showing beading
near their tips and ending in large
spherical globules 42
B. Nerve fibers in rostral stump of spinal
cord beginning their intrusion into the
scar at 20 days postoperative 42
C. Nerve fiber tips growing through the
scar at 30 days postoperative 42
5. Results of bouton counts as revealed
by the Rasmussen stain 49

LIST OF FIGURES
(Continued)
Figure
6.A. Polygraph tracing showing undulatory
movements caudal to transection site 55
B. Polygraph tracing showing response of
axial musculature following stimulation
of barbels of normal animal 55
7. A. Results of strength tests following
spinal cord transection 58
B. Results of timed swimming trials
following spinal cord transection 58
8. A. Comparison between synapse count on
motor horn cell bodies caudal to the
lesion and undulatory strength 67
B. Comparison between synapse count on
motor horn cell dendrites caudal to the
lesion and undulatory strength 67

Abstract of Dissertation Presented to the Graduate
Council of the University of Florida in Partial Fulfillment
of the Requirements for the Degree of Doctor of Philosophy
SPINAL CORD REGENERATION IN THE SHARK
by
John Bruce Gelderd
August, 1972
Chairman: Jerald J. Bernstein, Ph.D.
Major Department: Department of Neuroscience
The shark presents a unique central nervous system
for experimental analysis. The present series of experiments
assess the regenerative capacity of the central nervous
system in the nurse shark (Ginglymostoma cirratum).
The spinal cord was transected at the mid-thoracic
level in 28 nurse sharks. Four animals per group were
sacrificed at intervals of 10, 20, 30, 40, 60 and 90 days
postoperative. Two groups of fish underwent a subsequent spinal
cord retransection at the same site at 90 days postoperative and
were sacrificed 10 and 20 days later. Three sections of
spinal cord were removed from each shark for histological
analysis. A section of spinal cord at the site of lesion
was stained using a modified protargol silver stain to assess
regeneration across the site of lesion. Another spinal cord
section caudal to the lesion site was stained using a
viii

modified Nauta technique to show degenerating descending
nerve fibers. The Rasmussen stain was used for the light
microscopic demonstration of bouton terminaux on motor horn
cells caudal to the lesion. Behaviorally, timed trials for
swimming speed and a strength test for axial musculature
contraction caudal to the lesion site were performed at
five-day postoperative intervals.
Histological analysis showed a neuroglial-pial-
ependymal scar joining the stumps of spinal cord. Regeneration
across the site of lesion did not occur until 40 to 60 days
postoperative. Nerve fibers traversed the lesion site from
both stumps of cord and tended to follow blood vessels and
glial bridges within the scar. The number of descending long
tract nerve fibers reaching an area six spinal segments
caudal to the lesion was small (9-13%) . Despite this,
synaptic terminals on motor horn cells caudal to the lesion
showed an increase from 10 to 60 days postoperative (45% of
normal at 10 days postoperative to 92% of normal at 60 days
postoperative).
Immediately upon recovery from anesthesia, all
operated sharks exhibited undulatory movements caudal to the
site of transection while at rest which were independent of
volitional movements rostral to the lesion. These undulatory
IX

movements increased in strength up to 60 days postoperative
at which point they were statistically indistinguishable from
the normal strength of axial musculature. Swimming prowess
was markedly reduced following spinal cord transection and
was never recovered. Undulatory movements were uncontrollable
and proved detrimental to swimming ability. The body caudal
to the site of lesion remained paralyzed in normal attempts
to swim.
Retransection of the spinal cord at 90 days
postoperative showed no change in the strength of axial
musculature caudal to the lesion, timed trials or the number
of boutons on motor horn cells. Comparison between the increase
in undulatory strength and increase in synaptic contacts
on the motor horn cells caudal to the lesion showed a
high correlation (r=.93, PC.01).
It was concluded that the small amount of
regeneration across the site of lesion had no effect on the
swimming behavior of the operated sharks. The increase in
strength of undulatory movements was attributed to the
reestablishment of synaptic contacts on motor horn cells
caudal to the lesion by local, segmental sprouting.
x

INTRODUCTION
Historically, there have been many experiments
and theories aimed at the solution to the perplexing problem
of central nervous system regeneration. One of the earliest
investigators in this area was Ramon y Cajal, who
demonstrated that growth of central nervous system fibers in
embryos resulted from protoplasmic elongation rather than
from fusion of cellular elements (Ramon y Cajal, 1928, 1960).
Ramon y Cajal's conclusions raised the question whether
similar growth would occur following a transection of the
spinal cord of an adult animal. Subsequent studies were
undertaken in both mammalians and inframammalians to assess
anatomical and physiological regeneration of the central
nervous system.
1

MAMMALIAN STUDIES
Following the publications of Ramon y Cajal (1928)
and until the experiments of Sugar and Gerard (1940), little
was added to our knowledge of regeneration in the adult
mammalian spinal cord. During this period, numerous reports
appeared on the regeneration of fetal rat spinal cords.
These experiments met with little or no success and apparent
voluntary function was explained as spinal reflex activity.
Some abortive regeneration was seen, but the regenerated
fibers atrophied before becoming functional (Nicholas and
Hooker, 1928). In 1940, Sugar and Gerard succeeded in
obtaining structural and functional regeneration in the
transected spinal cords of young adult rats. In some of
these animals, muscle and nerve transplants had been oriented
in a longitudinal plane between the severed cord stumps.
These animals were able to sit up, clean their face,
responded by squealing when their tail was pinched and
exhibited coordinated locomotion. Stimulation of the cerebral
peduncles elicited vigorous movements of the hindquarters.
In experiments on spinal cats and dogs (Brown and
2

3
McCouch, 1947), the lack of functional regeneration in the
spinal cord was attributed to the formation of a dense
neuroglial-ependymal scar at the site of lesion. Further
study of this scarring phenomenon was done by Windle and his
associates (Windle and Chambers, 1950a, 1950b, 1951; Windle,
Clemente and Chambers, 1952; Windle, 1955, 1956; Clemente,
1955). After surgical transection of the spinal cord of
cats and dogs, a dense neuroglial cap or scar formed over
both cut ends of the spinal cord. This scar was composed
of fibroblasts and neuroglia. The relatively avascular
cicatrix between the two stumps of spinal cord was composed
of thick masses of collagenous connective tissue and
completely isolated the cut ends of the cord. It was this
scar which was thought to prevent regeneration and thus
inhibit return of function. Administration of the
bacterially derived polysaccharide, Piromen, prevented the
development of this dense neuroglial scar and permitted the
intrusion of blood vessels into the area of lesion. As a
result of this reduction of the scar, nerve fibers were able
to cross the loose cellular matrix that formed between the
two cut ends of the spinal cord. Freeman (1955) undertook
an extensive series of experiments concerning spinal cord
regeneration in rats. His experiments lasted for more than
15 years. During this time, spinal cord transection was

4
accomplished in over 7,000 rats. Anatomic regeneration and
normal functional return has occurred in approximately 100
rats. Electrophysiological studies on these rats showed
conduction of impulses in both directions through the site
of lesion. After complete functional recovery (six months
to one year), the "walking paraplegic" rats were injected
with procaine directly into the spinal cord at the site of
transection. The animal again became completely paraplegic.
Retransection of the spinal cord in such rats also returned
them to a paraplegic state. Freeman (1955), Littrell et al.
(1953) and Littrell (1955) transected the spinal cord in
adult cats and contrasted the recovery of non-treated animals
to those treated by intravenous injections of Piromen. Cats
treated with Piromen exhibited return of function beginning
at 2-3 months postoperative and peaked at 9-12 months. After
this point, the animals regressed to typical paraplegic
behavior by 18 months. Histology showed a dense neuroglial-
ependymal scar at the site of transection which was "choking
off" the regenerated fiber tracts. There was never any
return of sensory function. Non-treated animals did not
show any restitution of function or anatomical regeneration.
These studies indicated that the neuroglial-ependymal scar
prevented regeneration in mammals. However, this same
scarring phenomenon also occurred in inframammalian forms that

5
did indeed show both anatomical regeneration and
physiological return of function after spinal cord
transection. It was thought, therefore, that perhaps
mammalian central nervous system neurons lacked sufficient
growth potential to. regenerate lost peripheral processes.
Levi-Montalcini and Brooks (1960) tested the effects of a
protein isolated from the mouse salivary gland upon chick
and mouse sensory ganglia in vitro. Within 12 hours after
injection of this nerve growth factor (NGF) into the medium,
a dense halo of nerve fibers surrounded the explant.
Injection of NGF into intact animals produced hyperplasia
and hypertrophy of sensory and sympathetic neurons. Scott
and Liu (1964) injected NGF and Piromen into kittens after
crushing the dorsal columns. There appeared to be a
definite, direct correlation between the amount and duration
of NGF administration and the regeneration of the sensory
fibers. Anatomical regeneration across the site of lesion
was confirmed electrophysiologically in these kittens
although Scott and Liu did not wait for functional return.
Scott et al. (1966) administered NGF to young rats after dorsal
root crushing and found an increase of 14% in protein production
in dorsal root ganglia as compared to no increase without
administration of NGF. Harvey and Srebnik (1967) found
anatomical and physiological regeneration with return of function

6
following spinal cord compression in rats treated with
L-thyroxine. Non-treated rats showed no regeneration or
return of function. In another rare case of central nervous
system regeneration, Adams et al. (1968, 1969, 1971) cut the
infundibular stalk in ferrets. Degeneration occurred
initially, followed within two weeks by new fibers
regenerating from the hypothalamus through the fibrous scar.
These fibers carried neurosecretory material. During the
one-to three-month postoperative period, fibers grew to a
proximal ectopic infundibular process which formed following
the lesion. In animals kept alive until 12 months
postoperative, the entire neurohypophysis had been
reinnervated and was functional.
If a nerve fiber is severed, there appear to be at
least three distinct reactions: (1) growth does not occur
or it is abortive; (2) the original fiber may regenerate
and reform its synaptic contact or (3) adjacent intact
nerve fibers or cells may develop collateral sprouts and
reinnervate the deafferented tissue (Guth and Windle, 1970).
Liu and Chambers (1958) experimentally demonstrated sprouting
in the spinal cord of the cat following deafferentation
either by adjacent dorsal root section or by corticospinal
tract ablation at cranial levels. They observed that
paraterminal and collateral sprouting was rather generalized

7
in areas which had been deafferented and that the amount
of this sprouting was determined by the extent of the
denervation. Goodman and Horel (1966) showed restricted
sprouting of optic tract projections in the rat after
occipital cortex removal. Schneider (1970) lesioned the
visual cortex or superior colliculus in neonate and adult
hamsters. The neonate hamsters showed sprouting in optic
tract projections and some sparing in visual discrimination
tests in contrast to the adult hamsters which showed little
or no anatomical regeneration or return of function. By
using histochemical fluorescence techniques, Bjorklund and
his colleagues have shown regenerative axon sprouting in
the rat mesencephalon following electrolytic lesions
(Katzman et al., 1971; Bjorklund and Stenevi, 1971) and in
the rat spinal cord following spinal cord compression
(Bjorklund et al., 1971).
Raisman's investigations (1966, 1969a, 1969b)
in the septal nuclei of the adult rat constitute one of the
few ultrastructural studies on collateral sprouting in the
mammalian central nervous system. Afferents from two
separate pathways converge upon the medial septal nucleus.
Fibers originating in the hippocampus pass to the septum
through the medial forebrain bundle. Hippocampal fibers
terminate exclusively on the dendrites of the septal nuclei

-8-
whereas the hypothalamic fibers terminate primarily upon the
cell bodies of the septal nuclei. After lesioning the
hippocampal input, the remaining afferent septal fiber tract
showed a high proportion of axon terminals which made contact
with more than their normal share of postsynaptic units.
This phenomenon was interpreted as a reinnervation by the
remaining hypothalamic input of synaptic sites left open by
lesioning of the hippocampal input. This was corroborated
by lesioning the medial forebrain bundle which produced
degeneration of the remaining synaptic contacts. Raisman
then lesioned the hypothalamic input and found sprouting
of hippocampal fibers in the septum to fill synaptic sites
left by the degenerated hypothalamic input. Moore et al.
(1971) has recently duplicated Raisman's study utilizing
histochemical fluorescence techniques and has corroborated
Raisman's findings. Ultrastructural experiments on rat
spinal cord (Bernstein and Bernstein, 1971) have shown
similar results. Neurons were deafferented by hemisecting
the spinal cord. Shortly after these cells were deafferented,
they began to hypertrophy. In particular, the dendrites
exhibited profuse branching and irregular swelling. Synaptic
spines on the hypertrophied dendrites were also greatly
increased in number. Concomitant with this was the
establishment of large numbers of axodendritic synapses.

9
Furthermore, the axons that terminated near the site of
lesion appeared to arise by way of axonal sprouting from
the region of the spinal cord that was not hemisected. To
ascertain if there was any long tract involvement in the
regenerative process, a group of rats underwent hemisection
of the spinal cord just below T2. Ninety days later, the
spinal cord was hemisected on the same side at C5 and
the spinal cord at vertebral level T2 underwent histological
examination for degeneration. The region immediately rostral
to the T2 lesion showed new, degenerating nerve fibers in
small amounts. The area was also filled with degenerating
axodendritic synapses. These data suggested that the long
tracts were indeed partially involved in the regeneration of
axons to the neurons proximal to the site of the original
lesion. Bernstein stressed, however, that the number of
fibers appeared to be low and that the vast majority of new
synapses originated from segmental sprouting. This study
showed, however, that descending long tract nerve fibers in
the spinal cord of the rat are capable of limited
regeneration to the area immediately rostral to the lesion
site. The regenerating axons in the rat appeared to respond
to nonspecific influences of the hypertrophied dendrites
and established inappropriate connections. The formation
of these synapses then effectively terminated further growth

10
of the axons. Bernstein and Bernstein (in press) have also
shown limited regeneration of axons rostral to the site of
hemisection in the Rhesus monkey spinal cord. Motor horn
cell dendrites immediately rostral to the site of
hemisection showed varicosities. Regenerating axons made
normal as well as aberrant synaptic recombinations with
reactive neurons rostral to the lesion. The most frequent
type of aberrant synaptic complex was a cup-shaped bouton with
a central, large extracellular space between presynaptic and
postsynaptic membranes. In another recent ultrastructural
study, Lund and Lund (1971) found synaptic adjustment in the
superior colliculus following enucleation of neonatal and
adult rats. Little change was observed in the number and
types of synapses in neonatal rats due to synaptogenesis,
but adult rats showed a reinvasion of synaptic sites with
an incomplete return to a normal proportion of synaptic
types.
Although regeneration in the mammalian central
nervous system appears possible, functional return has been
shown in only isolated cases. The majority of experiments
have shown only abortive regeneration. In the instances of
functional return after spinal cord lesion, many are regressive
and the animal returns to a paraplegic state. Sprouting then
appears to be the rule in the mammalian central nervous system,

11
with functional regeneration the rare exception. There is,
as yet, no clear functional significance to the phenomenon
of central nervous system sprouting. McCouch et al. (1955)
has implicated sprouting in spasticity following spinal
cord transection and Schneider (1970) suggests that sparing
of pattern vision behavior in neonatal hamsters is due to
sprouting of visual pathways. However, this evidence is
only suggestive at best and more definitive studies are
required to shed more light on this anatomical phenomenon.

INFRAMAMMALIAN STUDIES
In contrast to the abortive regeneration found in
mammals, inframammalian forms have proven to be a fertile
area for successful central nervous system regeneration
studies. In fact, central nervous system regeneration seems
to be the rule in lower forms with abortive regeneration
the exception. Central nervous system regeneration in bird
embryos has been well documented (Clemente, 1955; Hamburger,
1955) although regeneration in the adult bird occurred only
in the visual system (Cattaneo, 1923). Central nervous
system regeneration studies in reptiles have centered
around the well known phenomenon of tail regeneration in
lizards (Clemente, 1964; Hamburger, 1955). The central
nervous system of amphibia has shown great regenerative
powers. With the noted exception of the adult Anurans, the
amphibia have proven comparable to teleosts in central nervous
system regeneration. After complete transection of the spinal
cord of larval salamanders (Piatt, 1955a, 1955b) or in the
axolotl (Kirsche, 1956), extensive regeneration of fiber
tracts was observed with a concomitant return of normal

13
function. In the adult salamander, regeneration was equally
vigorous (Piatt, 1955a, 1955b). Regeneration of the spinal
cord of the adult newt took place within 30 days with or
without injection of Piromen (Drummond, 1954). Regeneration
in the frog central nervous system has been restricted
to the larval stages (Hooker, 1925) . The adult frog has
shown abortive regeneration of the spinal cord after
transection (Piatt and Piatt, 1958; Clemente, 1964).
Regeneration in the frog and toad visual system, however,
has been very specific and successful, both anatomically
and physiologically (Sperry, 1944; Gaze and Jacobson, 1963;
Gaze and Keating, 1969, 1970a, 1970b; Gaze, 1970).
The regenerative capacity of the fish spinal cord
ranks high among the vertebrates. This has been shown
repeatedly through the almost exclusive use of the teleost
as an experimental animal in regeneration studies.
Regeneration in cyclostomes has been largely restricted to
spinal cord regeneration in larval forms (Marón, 1959;
Hibbard, 1963; Niazi, 1963). Spinal cord regeneration in
teleosts has proven so superior to the mammalian nervous
system that what would be considered poor functional or
anatomical recovery in teleosts would undoubtedly be hailed
as strikingly successful in man or any commonly used
laboratory mammal. Regeneration in teleosts does not occur

14
to the same degree in all parts of the central nervous
system. Regeneration throughout the central nervous system
in fish has recently been thoroughly reported by Segaar
(1965) and Bernstein (1970) and will not be repeated here.
The discussion here will be largely restricted to spinal
cord regeneration.
The regenerative capacity of the fish spinal
cord extends from the simple regrowth of axons across the
site of lesion to the complete restitution of neural
cytoarchitectonics, replete with new nerve cells and glia.
Koppanyi and Weiss (1922) carried out spinal cord transections
at a high level in goldfish. The fish were paralyzed caudal
to the lesion for two to three weeks, after which they began
to show signs of return of function. After 60 days, the fish
were behaviorally indistinguishable from normals. Histological
examination showed regeneration of neural pathways which
resulted in the reappearance of normal connections (Koppanyi
and Weiss, 1922; Koppanyi, 1955). Pearcy and Koppanyi (1924)
later cut the entire vertebral column of goldfish with
scissors so that no bony continuity remained between the
regions anterior and posterior to the section. Ten weeks
postoperatively, the fish were again swimming normally.
Hooker (1930, 1932) transected the spinal cord of guppies
less than four days old. He claimed full coordination and

15
integrative movements concomitant with the reestablishment
of nervous connections between the two halves of the body
approximately four days postoperatively. Keil (1940)
transected the spinal cord of adult rainbow fish and claimed
restitution of function beginning from three to twelve days
postoperative with complete restitution of function at
30-40 days postoperative. Ten years later, Kirsche (1950)
confirmed not only the functional but also the morphological
regeneration of the spinal cord in the adult rainbow fish.
Kirsche introduced the method of stimulating the spinal cord
above the site of transection. Tail fin movements were
elicited when the cord, and only the cord was stimulated
above the transection. Those animals which showed no
morphological regeneration also showed no movement of the
caudal fin upon stimulation of the spinal cord above the
transection. Kirsche distinguished various phases during
the course of regeneration. The first phase, which was
apparent approximately four days after the transection,
consisted of a disorganized growth from the severed stumps.
The second phase began about seven days postoperative with
a mitotic increase of the ependymal cells to form "indifferent
neural cells" which in time developed into neuroblasts and
glioblasts. Further differentiation lead to the formation
of normal cells in both proximal and distal stumps. Oriented

16
fibers grew out from these new cells and,approximately 15
days postoperatively, there was evidence for both morphological
continuity and functional recovery (Kirsche, 1950, 1965).
Healey (1962) transected the spinal cord of minnows and noted
the immediate inability of the minnow to change colors upon
background color reversal. Fast color changes were shown to
be under autonomic control and slow color changes were
mediated by hormonal control. Ten days after transection,
the fishes ability to change color increased. After four
months, rapid color changes occurred that were indistinguishable
from normal. Bernstein (1964) has shown a relationship
between age and regenerative capacity of the goldfish spinal
cord. Young goldfish (less than one year old) were able
to reconstitute approximately 90% of the available descending
axons whereas approximately 60% were reconstituted in two-
and three-year-old animals. The ability of the neuroglia
to regenerate and reestablish the diameter of the cord was
also age dependent. The younger goldfish reconstituted the
diameter of the cord almost completely (Bernstein, 1964).
Not only has the spinal cord of the teleost fish regenerated
after being severed, it also has the ability to completely
reconstitute areas of the spinal cord following ablation.
This type of growth pattern has been found in the regeneration
of the caudal neurosecretory system of Tilapia. After removal

17
of the caudal peduncle, tailfin and caudal spinal cord
segments, a new caudal neurosecretory system regenerated.
This system was somewhat abberant but fully functional
(Fridberg et al., 1966).
In a series of experiments on goldfish, it has been
shown that although there was return of function following
spinal cord transection, the morphological regeneration was
less than perfect (Bernstein, 1964; Bernstein and Bernstein,
1968; Bernstein and Gelderd, 1970; Bernstein and Gelderd,
in manuscript). Following spinal cord transection, goldfish
were paralyzed caudal to the site of lesion, descending spinal
tracts degenerated, and synaptic sites on perikaryon and
primary dendrites of motor horn cells 2 cm caudal to the site
of lesion dropped by 50%. Following 60 days regeneration
time, the synaptic complement on motor horn cells was
reestablished although descending fiber tract regeneration
was only 35-50% of normal. A subsequent retransection of the
spinal cord at 60 days postoperative resulted in degeneration
of the new, regenerated descending fibers and concomitant
paralysis caudal to the site of lesion. In contrast, there
was no statistically significant change in the synaptic
complement on motor horn cells 2cm caudal to the site of
lesion. This seemed to indicate that the descending fibers
regenerating into the caudal section of spinal cord did not

18
return to their original synaptic sites on motor horn cell
perikaryon or primary dendrites, but perhaps synapsed instead
on internuncial cells. Return of the normal synaptic complement
on motor horn cell perikaryon or primary dendrites was
relegated to local, segmental sprouting of adjacent fiber
tracts or cells. Those descending fibers which did not
regenerate past the site of lesion appeared to synapse on
cells near the lesion site. When the regenerating axons
reached the site of lesion, they were confronted with
deafferented neurons. The regenerating axons synapsed on
these cells until the synaptic sites were filled. Once the
regenerating axons made these inappropriate synaptic contacts,
they ceased their growth. The mechanism for the cessation
of growth of these axons is thought to be a special case of
contact inhibition (Bernstein and Bernstein, 1968). It must
be stressed again that regeneration in the goldfish central
nervous system is not a uniform phenomenon. Although
transection of the spinal cord was followed by a reduced
regenerative capacity in the number of fibers traversing the
lesion (Bernstein and Gelderd, 1970), lesions of the visual
system were followed by a more specific regenerative process
which initially appeared to be point for point with 100% of
the original optic fibers regenerating (Attardi and Sperry,
1963; Jacobson and Gaze, 1965). Recent studies, however,

19
have shown some deviation in the area of termination of
regenerating optic nerve fibers in the optic tectum of the
goldfish following lesions in the visual system (Yoon, 1971;
Horder, 1971; Sharma, 1972).
In mammalians, the neuroglial-ependymal scar is
thought to be responsible in part for the apparent lack of
regeneration in the central nervous system, forming a dense
barrier between tie cut ends of the spinal cord and thus
preventing regeneration (Brown and McCouch, 1947; Windle,
1955; Windle and Chambers, 1950a, 1950b; Guth and Windle,
1970). Bernstein and Bernstein (1967) investigated the
effect of the neuroglial-ependymal scar on spinal cord
regeneration in goldfish. The spinal cord was completely
transected and a thin teflon disc was placed between the two
cut ends of the spinal cord. The teflon disc remained
between the cut ends for 30 days which was ample time for
regeneration to occur in goldfish. The goldfish were then
operated upon again and the teflon disc removed. These
goldfish were observed for an extended period of time to
determine any return of function caudal to the lesion which
would signify regeneration of the spinal motor tracts. No
regeneration or return of function was observed six months
later. Other goldfish were operated upon again at 30 days
postoperative. The teflon disc was removed and the spinal

20
cord transected one spinal segment rostral to the original
lesion. After 30 days, the spinal cord was observed
histologically to assess regeneration. The descending
spinal motor tracts grew through the second lesion, caudalward
through the isolated- section of the spinal cord and through
the first lesion which was delineated by a substantial
neuroglial-ependymal scar. Hence, the regenerative capacity
of the goldfish spinal cord was not affected by the
neuroglial-ependymal scar acting as a mechanical barrier
(Bernstein and Bernstein, 1967) .

BLASMOBRANCH STUDIES
The shark presents a unique nervous system, both
among the fishes specifically and vertebrates in general.
One trait which is particularly unique to the elasmobranchs
is the reported absence of internuncial cells (Golgi Type II)
in the spinal cord (Kappers et al., 1936; Von Lenhossek,
1892, 1895; Aronson, 1963; Nieuwenhuys, 1964). Presumably,
the vast majority of descending tracts end directly on
motor horn cells without intermediary neurons to intercede or
modulate information from higher centers. Another unique
characteristic restricted to the elasmobranchs is the high
urea content in the blood and sera (Goldstein, 1967; Goldstein
et al., 1968; Rasmussen and Rasmussen, 1967; Smith, 1929;
Rasmussen, 1971). This phenomenon is particularly evident
in marine elasmobranchs. This large concentration of urea
is also reflected in the cerebrospinal fluid (Smith, 1929;
Rasmussen, 1971) and may have a profound effect on the
regenerative capacity of the shark central nervous system
because urea is used to shrink brain tissue during neurological
operations.
21

22
In addition to the above-mentioned characteristics
of the shark central nervous system, this class of fish
shows important advancements in the evolution of the
vertebrate central nervous system. The structure of the
spinal cord in elasmobranchs may be considered a prototype
for that of high vertebrates. Unlike the spinal cord of
cyclostomes which is flat, the spinal cord of elasmobranchs
is round or oval. In addition, these are the first primitive
animals to have myelinated fibers in the spinal cord and
whose dorsal and ventral roots unite outside the vertebral
column to form a mixed root. It is also in this class that
one first finds the division of the gray matter into dorsal
and ventral horns and the first time that all cells of
origin for sensory fibers in the cord lie in extramedullary
spinal ganglia (Kappers, 1906; Kappers et al., 1936; Aronson,
1963; Nieuwenhuys, 1964).
In mammals and other inframammalian forms,
transection of the spinal cord produces paralysis immediately
after transection. In most inframammalians, paralysis is
alleviated by regeneration and return of function after
varying periods of time. In the case of mammals, spinal
walking may occur after weeks or months of recuperation.
The shark, however, has exhibited coordinated undulatory
movements immediately upon recovery from anesthesia after

23
spinal cord transection (Ten Cate and Ten Cate-Kazejawa,
1933; Gray and Sand, 1936a, 1936b; Lissmann, 1946a, 1946b;
Healey, 1957) and was able to swim, using coordinated
movements between those portions of the body rostral and caudal
to the transection. Ten Cate and Ten Cate-Kazejawa (1933)
removed all the muscles in the region of the anterior dorsal
fin of the dogfish (Scyllium canícula and S. catalus) and
transected the spinal cord in the same region. He observed a
locomotory rhythm propogated over the site of the operation,
thereby maintaining coordinated movement between the head and
posterior region of the body. According to Ten Cate and Ten
Cate-Kazejawa (1933), the activity of the posterior region
of the body depended upon tensile stimuli applied to the
posterior musculature whenever an active contraction occurred
in the head region. If this conclusion is justified, then
the normal locomotory rhythm of the dogfish involves the
activity of a chain of peripherally controlled reflexes.
Gray (1936) and Gray and Sand (1936a, 1936b) disagreed with
the hypothesis of Ten Cate and Ten Cate-Kazejawa. They
showed that coordinated responses no longer occurred if two
regions of the body of a dogfish were isolated from one
another by a second spinal cord transection. If both of
these regions were of sufficient length, each exhibited an
independent, spontaneous, automatic activity within the

24
spinal cord. Lissman (1946a, 1946b) showed that locomotory
rhythm could only be abolished in spinal dogfish by a
complete, bilateral rhyzotomy caudal to the transection.
This conclusion appeared to support Ten Cate's hypothesis.
The continued locomotory rhythm after spinal cord transection
in sharks may in part be attributed to its rather low
position on the phylogenetic ladder. Eels have shown an
undulatory behavior after decapitation, having a duration of
only a few seconds (Gray, 1936). Gray has attributed this
phenomenon to injury potentials in the remaining spinal
cord. Typically, spinal eels laid on their side and showed
no coordination between rostral and caudal portions of the
body in normal attempts to swim. Nociceptive stimuli did,
however, cause undulations (Gray, 1936). Little mention has
been made of swimming prowess in the shark with respect to
strength and speed after spinal cord transection. An attempt
to clarify these questions was made during this series of
experiments.
Central nervous system regeneration studies
in elasmobranchs are conspicuous by their absence. Although
lesion studies have been done (Ten Cate and Ten Cate-
Kazejawa, 1933; Gray and Sand, 1936a, 1936b; Lissman,
1946a, 1946b; Healey, 1957; Segaar, 1965; Ebbesson, 1972),
there have been no known experiments concerning

25
shark central nervous system regeneration. Indeed, experimental
work per se on the shark central nervous system is at best
limited. It is this complete lack of central nervous system
regeneration experiments on sharks plus the anatomical and
behavioral uniqueness of their central nervous system that
have prompted me to use them as experimental animals. It was
my intent to use these unique features of the shark central
nervous system to shed some light on the regenerative
process.

EXPERIMENTAL
Materials and Methods
Subjects
Thirty-six male and female nurse sharks
(Ginglymostoma cirratum), approximately two feet in length,
were used. These sharks were trapped in the coastal waters
off Ft. Lauderdale, Florida.
Environment
All fish were kept at Marineland of Florida in an
outside, circular, salt water tank 15 feet in diameter and
six feet in depth. A constantly circulating salt water
system was used to insure proper oxygenation, salinity and
water temperature. Sharks were fed to satiation daily on
cut-up fish.
Operative Procedures
All operated fish were anesthesized with Tricaine
methanesulfonate (MS-222, 1:4000, Finquel, Ayerst Laboratories),
then placed on an operating board. A longitudinal incision
was made at the midline in the midthoracic region at the level
of the trailing edge of the pectoral fin, and the musculature
26

27
dissected away to expose the spinal column. A laminectomy
was performed and the spinal cord transected with a scalpel.
The wound was sutured and powdered sulfathiazole-sulfonilimide
was applied to the suture line to prevent infection. Following
surgery, all animals were also given a 0.1 cc intramuscular
injection of Longicil. Animals were tagged for identification
by attaching a numbered clamp and colored streamers to the
anterior dorsal fin. All sharks, including four normals,
were separated into nine groups (four animals per group)
and were killed by anesthetizing them at 10, 20, 30, 40, 60
and 90 days postoperative and perfusing them with 10% buffered
formalin. Two groups of animals underwent a subsequent
retransection of the spinal cord at the same site at 90 days
postoperative and one group each was killed by the above
fixation method at 10 and 20 days postoperative (Table 1).
Histology
Three sections of the spinal cord were removed
from each shark: a 2 cm section at the site of lesion, a
second section (1 cm in length) six spinal segments caudal
to the site of lesion and a 1 cm section immediately caudal
to the second section (Fig. 1).
The 2 cm section of spinal cord at the site of
lesion was sectioned horizontally at 15q and stained using
a modified Bodian silver technique counterstained with

TABLE 1
SUMMARY OF EXPERIMENTAL PARADIGM
Group
dumber
Number
of
Animals
Spinal
Cord
Transection
Sacrifice Day
Following
Transection
Bodian
Stain
(Nerve Fiber)
Nauta
Stain
(Deqen. Fiber)
Rasmussen
Stain
(Synapses)
Behav. Tests
Five Day
Intervals
; 1
Normal
4
No
1
N/A
X
Normal
X
Normal
X
Normal
2
4
Yes
10
X
X
X
X
3
4
Yes
20
X
X
X
X
4
4
Yes
30
X
X
X
5
4
Yes
40
X
X
X
6
4
Yes
60
X
X
X
7
4
Yes
90
X
X
X
8
4
Yes
10*
X
X
X
X
9
4
Yes
20*
X
X
X
X
TOTAL 36
• NOTE: These fish underwent a subsequent spinal cord retransection at the same site at 90 days
postoperative. This number indicates the sacrifice day following the second
transection.

Figure 1
- Location of spinal cord sections removed for
histological analysis.

30
Spinal Cord
Section Taken
for Bodian
Site of
Transection
Spinal Cord
Section Taken
for Nauta Stain
Spinal Cord Section
Taken for Rasmussen
Stain
6 spinal segments

31
cresyl-violet and eosin. The regenerative process was
assessed with respect to (1) formation of a neuroglial-
ependymal scar, (2) the effect of the scar on regeneration
and (3) the rate of the regenerative process.
Three descending tracts (tectospinal, thalamospinal
and ventral cerebellospinal tract) were studied in the 1 cm
segment of spinal cord located six spinal segments caudal
to the site of lesion. This section of spinal cord was
histologically analyzed with respect to the number of
degenerating nerve fibers within a given tract following
spinal cord transection (Fig. 1). The spinal cord was
serially sectioned horizontally at 30q and impregnated
using a modified Nauta technique for degenerating nerve
fibers. The degenerative pattern was plotted by drawing a
composite cross-sectional diagram made by examining, in order,
each horizontal section. The histological sections
corresponding to the known anatomical locations of the
given descending tracts were selected and the number of
degenerating fibers counted in each tract at 10 and 20 days
postoperative. These data were compared to the number of
degenerating nerve fibers found 10 and 20 days following
the subsequent retransection (Table 1). The second transection
controlled for local versus long tract input following
regeneration.

32
A quantitative analysis of the synaptic terminals
was done on the perikaryon and primary dendrites of ventral
motor horn cells following spinal cord transection. The
section (1 cm in length) of spinal cord used was taken
immediately caudal to the section used for fiber tract
analysis so that the number of regenerating fibers and the
synaptic profiles could be compared (Fig. 1). The spinal
cord was sectioned coronally at 10q and impregnated using
the Rasmussen stain for the light microscopic demonstration
of bouton terminaux (Rasmussen, 1957) and followed by a
cresyl-violet and eosin counterstain. Counts were made on
only those motor horn cells in which a prominent nucleolus
and primary dendrite could be seen in a given section.
Counts were made on a total of 576 motor horn cells. Sixteen
motor horn cells were counted per shark, utilizing eight
cells on the left side and eight cells on the right side.
Synaptic terminals were counted at 10, 20, 30, 40, 60, and 90
days postoperative and on the two groups of fish retransected
at 90 days postoperative (Table 1). All counts were made on
coded slides to insure unbiased results. The resultant data
were decoded and the levels of significance determined for
intra- and intergroup interactions by using computer
program BMDX63 for multivariate analysis of variance.

33
Behavior
A behavioral analysis of the sharks was done
during the postoperative period including the two groups
retransected at 90 days. The operated sharks were observed
daily while swimming in the tank and compared to normal
sharks with respect to swimming prowess. In addition, two
quantitative tests were performed on all sharks,
preoperatively and at five-day postoperative intervals.
The first test consisted of removing each shark from the
tank and strapping it to a board with that portion of the
body rostral to the lesion firmly held in place. That
portion of the body caudal to the lesion remained unrestrained
with the exception of the caudal peduncle to which a hose
clamp was attached. The clamp was connected by way of a
#10 screw to a Statham load cell assembly (Model UL-4)
which was in turn mounted on a Statham universal force
transducer (Model UC-3). The entire transducer assembly
was securely mounted on the test board. The output of the
force transducer was fed into a two-channel Grass polygraph
recorder. Two electrodes were attached to the paired
barbels located on the underside of the snout of the shark
(Fig. 2). The shark was stimulated using a constant current
stimulator producing a 10 ma pulse of 50 msec duration.
The strength of the response of the caudal body musculature

Figure 2
- Apparatus for testing strength of axial
musculature caudal to the site of spinal
cord transection.

35

36
following stimulation was recorded on the polygraph and
compared to preoperative and normal data. A minimum of
five responses was recorded for each shark on a given
trial day.
The second test consisted of timed swimming
trials. Each shark was placed in the water at one end of a
7' x 3' x 2' tank and the time required for the shark to
swim the length of the tank was recorded. Two consecutive
timed trials were measured on each shark on a given trial
day. Only those trials were counted in which swimming was
uninterrupted over the entire distance. The postoperative
data for both behavioral tests were compared to normal data
in addition to intra- and intergroup interactions among
postoperative groups by utilizing computer program BMD08V
for analysis of variance.

RESULTS
• Histological
Site of Lesion
Histological analysis at the site of lesion in
the group of animals sacrificed at 10 days postoperative
showed a dense scar separating the cut ends of the spinal
cord (Fig. 3A). The diameter of the cord in the scar area
was approximately 75% of normal. The scar was composed of
neuroglial, ependymal and pial cells. Blood cells and
phagocytes were also present in abundance throughout the
scar, but no blood vessels were seen within the scar at
this time. Three types of cisterns were found within or
near the site of lesion. A cistern lined with ependymal
cells was visible at the rostral stump of spinal cord.
Several smaller cisterns lined with endothelial cells
were also present throughout the scar. A third type of
space or cistern was found near the lesion site in both
stumps of spinal cord. These spaces were not lined with
cells and some had severed nerve fiber tips within them.
All three types of cisterns were seen in the lesion

Figure 3 - Site of spinal cord transection showing cistern
lined with ependymal cells <☆> , at rostral
stump of cord, small cisterns within the scar
(^r) lined with endothelial cells and cisterns
in both stumps of spinal cord ( | ) some of which
enclose severed tips of nerve fibers. Bodian
silver stain (X10).
A. 10 days postoperative
B. 30 days postoperative
C. 90 days postoperative - cut ventral root
growing from rostral stump of cord into
scar area (f)•

39

40
area during the entire postoperative period (Fig. 3A, 3B,
3C). No nerve fibers were found within the scar at this
time. Many large nerve fiber tips were, in fact, found well
back from both cut ends of spinal cord (Fig. 3A). These
large nerve fibers, were beaded at their terminals and ended
in large spherical globules (Fig. 4A). This phenomenon
persisted throughout the postoperative period.
At 20 days postoperative, the scar area resembled
that at ten days. Nerve fibers were seen immediately adjacent
to the scar with some fibers beginning their intrusion into
the scar from both stumps of cord (Fig. 4B).
At 30 days postoperative the cistern lined with
ependymal cells had increased considerably in size and the
diameter of the scar was further reduced (55% of normal).
Nerve fibers were seen penetrating the scar from both
stumps of cord (Fig. 4C) but the center of the scar was
still devoid of nerve fibers.
At 40 days postoperative there was little change
in the appearance of the scar or the density and intrusion
of nerve fibers into the scar area.
There were no discernable differences in the
appearance of the lesion site between 60 and 90 days
postoperative. The scar appeared as a loose cellular matrix
made up of neuroglial and ependymal cells. The ependyma-

Figure 4
A. Large severed nerve fibers showing beading
near their tips and ending in large spherical
globules (|). Bodian silver stain (XI,000).
B. Nerve fibers in rostral stump (RS) of spinal
cord beginning their intrusion into the scar
(S) at 20 days postoperative. Bodian silver
stain (X450) .
C. Nerve fiber tips growing through the scar at
30 days postoperative. Bodian silver stain
(XI, 000) .

42

43
lined cistern at the rostral stump of cord had increased
still more in size (Fig. 3C) but no further constriction
in the diameter of the scar was observed. The increase in
the size of the ependyma-lined cistern in the rostral stump
of spinal cord was probably due to blockage of the flow of
cerebrospinal fluid in the central canal of the spinal cord.
There was a large increase in the number of nerve fibers
within the site of lesion. These nerve fibers were of
small caliber and appeared to completely traverse the scar
by following neuroglial bridges and blood vessels. In those
sharks where the transection was made immediately caudal to
the emergence of a pair of ventral roots, the ventral roots
were severed in the process of transecting the cord.
Severed ventral roots were seen, at 60 and 90 days
postoperative, growing caudally along the edge of the cord
until they reached the lesion site where they grew into the
scar and the caudal stump of spinal cord (Fig. 3C). Nerve
cell bodies were not found within the site of lesion at any
time during the postoperative period.
Nauta Stain
In the process of counting the number of fibers
in the three descending tracts following spinal cord
transection, it was determined that the tectospinal and
thalamospinal tracts could not be effectively counted

44
separately because they were anatomically adjacent to one
another within the spinal cord. Consequently, they were
counted as one tract. There was little evidence of
degeneration 10 days postoperative to both the first
transection and the subsequent retransection. This agreed
with lesion studies in the shark visual system utilizing the
Nauta technique where signs of degeneration following
lesions did not occur until approximately 20 days
postoperative (Ebbesson and Ramsey, 1968; Ebbesson and
Schroeder, 1971) . Those sharks sacrificed at 20 days
following both transections showed degeneration within the
three descending tracts and were thus used to make the counts.
Degenerating descending nerve fibers showed typical
irregular beading and droplet formations with concomitant
phagocytic activity typical of lower vertebrates. Ascending
tracts showed no retrograde degeneration.
Nerve fiber counts in the ventral cerebellospinal
tract and the combined tectospinal-thalamospinal tracts are
summarized in Table 2. Degenerating nerve fibers counted
20 days following the first transection represent the normal
complement of axons in each of the respective tracts. Those
nerve fibers counted 20 days following the retransection at
90 days postoperative were the number of regenerated axons
that originated rostral to the lesion site. The number of

45
TABLE 2
SUMMARY OF DEGENERATING NERVE FIBER
COUNTS IN DESCENDING TRACTS
Tract
Numbers
of Fibers
Normal
Regenerated
90 Days
% Regenerated
90 Days
Ventral Cerebellospinal
774
72
9.3
Tectospinal-Thalamospinal
1584
213
13.4

46
degenerating nerve fibers found within the combined
tectospinal-thalamospinal and ventral spinocerebellar tracts
following the second transection were 13.4% and 9.3% respectively
of the number of degenerating nerve fibers found following the
first transection.
Rasmussen Stain
The results of the synaptic terminal counts as
revealed by the Rasmussen stain on cell bodies and primary
dendrites of motor horn cells caudal to the lesion are
summarized in Table 3 and Fig. 5. The average number of
boutons represented was the combined data on both the left
and right sides of the spinal cord since there were no
statistically significant differences (P>.05) between
right and left counts. In addition, there was a high
correlation (r=.974, P_<.01) between bouton counts on cell
bodies and primary dendrites.
The number of boutons on cell bodies were
statistically less than normal (£<.05) throughout the
postoperative period, including the retransected groups.
There was, however, a statistically significant increase
in boutons with time after surgery from 10 to 60 days
(P.<.05) , but no statistically significant difference occurred
between 60, 90 and the two retransected groups (P^>.05) .
Synaptic terminals dropped to 45% of normal at 10 days

47
TABLE 3
SUMMARY OF’ COMBINED LEFT-RIGHT BOUTON COUNTS
ON MOTOR HORN CELL BODIES
AND PRIMARY DENDRITES
Days
Postoperative
Combined
Left-Right
Cell Body
Count
(XJ_SEM)
%
of
Normal
Combined
Left-Right
Count Per 10i^
Primary Dendrite
(X+SEM)
%
of
Normal
10
14.15+0.82
45.06
5.30+0.49
62.27
20
17.50^0.47
54.61
5.091,0.98
59.81
30
20.50^0.72
65.28
5.92,10.42
69.56
40
23.86^0.25
75.98
6.58.+0.17
77.32
60
28.96l_0.32
92.22
7.30.+ 0.38
85.78
90
29.90^0.12
95.22
7.78.+ 0.37
91.42
RETRANSECTION AT
90 DAYS
POSTOPERATIVE
10
29.90i_0.49
95.22
7.421,0.39
87.19
20
29.78^0.49
94.84
7.80+0.27
91.65
Normal
31.40+0.48
8.51+0.15

Figure 5 - Results of bouton counts as revealed by the
Rasmussen stain. There was a high correlation
(r=.97, P <;01) between counts on motor horn
cell bodies and primary dendrites.

NUMBER OF BOUTONS CELL BODY
49
z
o
-o
*
>
JO
<
z
O
DAYS POSTOPERATIVE
NUMBER OF BOUTONS PER 10M

50
postoperative, increased to 92% of normal by 60 days
postoperative and were 95.2% of normal at 90 days
postoperative. The retransected groups sacrificed at 10 and
20 days following retransection were 95.2% and 94.8% of
normal respectively.-
The number of boutons per lOg primary dendrite
following spinal cord transection were significantly less
than normal from 10 to 60 days postoperative (P<.05) and
were significantly greater (P (62.2% of normal at 10 days, 85.7% of normal at 60 days).
The bouton count was 91.4% of normal and statistically
indistinguishable from normal (P>.05) at 90 days
postoperative. Ten days following the retransection,
however, the number of boutons was 87.2% of normal and
statistically less than normal (P<,05). In contrast, there
was no statistically significant difference between normals
and 20 days following the retransection (P>.05).

BEHAVIOR
Daily Observations
Immediately upon recovery from anesthesia, all
operated sharks exhibited undulatory movements caudal to
the site of spinal cord transection. This phenomenon
persisted throughout the postoperative period and was never
observed in normal animals. Although these undulatory
movements occurred caudal to the lesion, there were no
swimming movements caudal to the lesion when the animals
attempted to swim. Forward movement was accomplished by
"walking" along the bottom of the tank using the pectoral
fins or by jerking the body rostral to the lesion left and
right while dragging the caudal portion of the body. Turning
could only be done by walking movements of the pectoral fins.
The undulatory movements during the early
postoperative days (1-30 days) were not strong enough to
move the operated animals. From 30 days postoperative,
however, undulatory movements became strong enough to
propel the sharks forward. The animals did not appear
able to control these undulatory movements. As a result,
51

52
the undulatory movements proved detrimental to their swimming
ability. Stimulation caudal to the lesion site by gentle
prodding or by an inadvertant touch by another shark caused
an increase in undulatory strength which either flipped the
shark over on its back, using the snout as a pivotal point,
or pushed the startled animal into a wall despite its best
efforts to prevent this by using the pectoral fins to
"backpedal" away from the wall. The sharks remained
paralyzed caudal to the transection for the duration of the
postoperative period with respect to normal attempts to swim.
Quantitative Tests
The results of the strength tests and timed swimming
trials are summarized in Table 4.
The strength .tests (Fig. 2) showed two types of
responses. The first response was elicited following
stimulation and was in the form of a single, sharp flexure
of the axial musculature. A second, consecutive stimulation
elicited the same type of response in the opposite direction
(Fig. 6B). This response following stimulation was present
in both normal and operated animals, although much reduced
in the operated animals. This response in operated animals
was probably due to passive conduction of movements caused
by muscle contractions rostral to the lesion. The second
type of response was the previously mentioned undulatory

53
TABLE 4
SUMMARY OF STRENGTH TEST RESULTS
FOLLOWING SPINAL CORD TRANSECTION
UNDULATORY
STRENGTH
DAYS IN KGMS
POSTOPERATIVE (X±SEM)
STRENGTH OF
RESPONSE
FOLLOWING
STIMULATION
IN KGMS
(XiSEM)
%
OF
NORMAL
TIMED
SWIMMING
TRIALS
IN SECONDS
(X+SEM)
°/o
OF
NORMAL
1
0.08±0.01
0.32±0.04
10.63
9.30± 0.30
16.12
10
0.21±0.03
0.29±0.03
9.63
8.86± 0.27
16.70
15
0.21*0.02
0.3210.04
10.63
8.73± 0.44
16.95
20
0.26+0.02
0.2710.03
8.97
8.89-0.38
16.64
25
0.37+0.04
0.3410.04
11.29
9.84+0.47
15.04
30
0.47± 0.06
0.31±0.02
10.29
8.70±0.44
17.01
35
0.87± 0.12
0.3510.03
11.62
8.06±0.37
18.36
40
1.14± 0.12
0.3810.04
12.63
7.85±0.30
18.85
50
1.77± 0.18
0.4810.05
15.94
7.84±0.28
18.87
55
1.96+0.24
0.5010.05
16.61
8.43±0.36
17.55
60
2.66± 0.30
0.4910.03
16.27
8.44+0.20
17.53
65
2.13-0.23
0.3410.02
11.29
8.73±0.31
16.95
70
2.23+0.21
0.3510.02
11.62
8.17±0.38
18.11
75
2.62± 0.63
0.3610.05
11.96
9.17± 0.55
16.13
80
2.67±0.20
0.3510.03
11.62
8.71+0.32
16.99
85
2.60±0.18
0.3610.01
11.96
9.08± 0.19
16.29
90
2.4610.18
0.3810.02
12.63
9.03± 0.09
16.38
RETRANSECTION
AT 90 DAYS POSTOPERATIVE
1
2.2810.16
0.3110.01
10.29
10.45± 0.17
14.16
10
2.3210.19
0.3110.02
10.29
9.75±0.32
15.17
20
2.4410.18
0.3210.01
10.63
9.66± 0.26
15.32
Normal
N/A
3.01± 0.32
1.48 0.22

Figure 6
A. Polygraph tracing showing undulatory movements
(0.25-0.5 cycles/sec) caudal to transection
site. The top trace indicates when stimulation
occurred. Paper speed = 3mm/sec.
B. Polygraph tracing showing response of axial
musculature following stimulation of the barbels
of a normal animal. The top trace indicates
when stimulation occurred. Paper speed = 3mm/se

55

56
movements caudal to the site of lesion. These undulatory
movements occurred only in operated animals and were an
almost constant phenomenon requiring no stimulation
(Fig. 6A) .
The resppnse following stimulation was significantly
reduced from normal throughout the postoperative period
001) following the first transection and there was no
trend during this postoperative period for any return of
strength caudal to the lesion. There was also no statistically
significant difference (P>.05) between the response following
stimulation at 90 days postoperative and the retransected
groups (Fig. 7A) .
Undulatory movements (0.25-0.5 cycles/sec) were
weak during the early postoperative period, but there was a
statistically significant increase in the strength of
undulatory movements with increased time from 20 to 60 days
postoperative (P_<.05) . From 60 days postoperative to 20
days following the retransection, there were no significant
differences between postoperative groups (P>.05). In addition,
the strength attained by undulatory movements at 60 days
postoperative was statistically indistinguishable (P>.05)
from the strength of response following stimulation of
normal animals (Fig. 7A).
A significant increase was observed in the time

Figure 7 -
A. Results of strength tests following spinal
cord transection.
B. Results of timed swimming trials following
spinal cord transection.

58
*
o
<
m
*
m
Z
—i
<✓»
B
STRENGTH IN KGMS OF UNOULATORY

59
required to swim seven feet following spinal cord transection
(P_<.001) . Normal animals required an average of 1.48 seconds,
whereas the mean value for transected animals was 8.72
seconds. There was no trend during the postoperative period
for improvement in swimming times (Fig. 7B) and there were
no significant differences between times following the
first and second spinal cord transections (P>.05).

DISCUSSION
These results indicate that following spinal cord
transection, the nurse shark is capable of limited anatomical
regeneration of descending tracts across the site of lesion
(9.3-13.4%) to an area six spinal segments caudal to the
lesion. The time required for nerve fibers to regenerate
across the lesion site was between 40 and 60 days. This was
considerably slower and more incomplete than regeneration in
the teleost spinal cord. Teleosts have shown anatomical
regeneration and return of normal swimming function from
four days postoperative in guppies (Hooker, 1930, 1932)
to approximately 35 days postoperative in goldfish (Bernstein,
1964) although descending fiber tracts in the goldfish
regenerate only 35-49% of the original complement of fibers
within a given tract (Bernstein and Gelderd, 1970).
Limited regeneration did occur in the shark but there was
no return of strength in the axial musculature caudal to
the spinal cord transection following stimulation rostral
to the lesion. The operated sharks were paralyzed caudal
to the lesion in normal attempts to swim. This lack of
60

61
strength caudal to the lesion was also reflected in the
timed swimming trials as there was no trend during the
postoperative period for improvement in swimming times.
In fact, the small amount of anatomical regeneration seen
six spinal segments caudal to the lesion at 90 days
postoperative had little or no effect on strength or
swimming speed as a subsequent retransection at 90 days
postoperative caused no change in these performance parameters
in the ensuing postoperative period.
As was stated previously, only a small number of
nerve fibers regenerated rostro-caudal across the site of
lesion to the area six spinal segments caudal to the lesion
and they did not traverse the lesion site until 40 to 60
days following transection. Despite this, the number of
synapses on motor horn cell bodies and primary dendrites
caudal to the lesion showed an increase beginning at 20 days
postoperative through 60 days postoperative and were highly
correlated (r=.974, P_<.01) .
If synaptic return occurs before the return of
long tract input, what is the source of the increase in the
number of synapses on motor horn cells caudal to the lesion?
The high correlation between primary dendrite and cell body
synaptic counts strongly indicates that both phenomena have
the same origin. The origin for the nerve fibers which

62
replaced lost synaptic contacts must have been caudal to
the site of lesion in the form of local, segmental sprouting.
This hypothesis is further supported by data following the
retransection at 90 days postoperative. There was no
significant change in the number of boutons on motor horn
cell bodies following retransection and a small but statistically
significant (P<.05) drop (91.4% to 87.6% of normal) in boutons
on motor horn cell primary dendrites. This slight drop in
boutons on primary dendrites following retransection was
probably due to degeneration of the small number of regenerated
long tract nerve fibers which synapsed on motor horn cell
dendrites.
If there is no return of swimming prowess and no
return of axial musculature strength following stimulation,
then the question arises as to the functional significance
of this sprouting phenomenon and the return of synaptic
contacts on motor horn cells caudal to the lesion site.
Perhaps the answer to this question lies in the unique
appearance in sharks of undulatory movements caudal to the
lesion following spinal cord transection. Before this
relationship is discussed, however, the anatomical basis
for the undulatory movements will be elaborated.
Unlike other shark studies which claim coordinated
undulatory movements propagated rostro-caudally over the

63
site of lesion (Ten Cate and Ten Cate-Kazejawa, 1933;
Gray, 1936; Gray and Sand, 1936a, 1936b), undulatory movements
in this experiment were observed only caudal to the lesion
site and were independent of body movements rostral to the
lesion. In fact, during the strength tests, the response
following stimulation was often superimposed upon undulatory
movements without affecting the speed or strength of the
undulatory movements.
There are at least two hypotheses relating to the
anatomical basis of undulatory movements in spinal sharks.
Ten Cate and Ten Cate-Kazejawa (1933) claim that the
undulatory movements are propagated over the lesion site
by tensile stimuli applied to posterior musculature when an
active contraction occurs in the head region, implying the
activity of a chain of peripherally controlled reflexes.
Gray (1936) and Gray and Sand (1936a, 1936b) showed that
coordinated responses did not occur if two regions of the
body of a dogfish were isolated from one another by a
second spinal cord transection. Each isolated section of
the body exhibited a spontaneous, independent undulatory
activity. Gray and Sand attributed this to an inherent
undulatory discharge rhythm within the spinal cord.
Lissman (1946a, 1946b) has shown that undulatory movements
caudal to a spinal cord transection in dogfish can only be

64
abolished by a complete bilateral rhyzotomy caudal to the
lesion. The hypothesis of Ten Cate and Ten Cate-Kazejawa
(1933) must be rejected in the present experiment because
undulatory movements persisted without movement rostral to
the lesion. In fact, the undulatory movements were most
prevalent when the shark was at rest on the bottom and were
absent in normal attempts to swim. The two theories need
not be mutually exclusive, however. Lissmann's studies
showed that dorsal root input is mandatory for the maintenance
of undulatory movements. Thus it appears from the present
data that the spinal cord of the shark has an inherent
undulatory discharge pattern modulated by local sensory
input and input from brain centers. If the major effect
of the brain on this undulatory discharge pattern is
inhibitory, transecting the spinal cord will release the
caudal section of spinal cord from these inhibitory influences,
thus allowing the inherent discharge pattern to be exhibited
in the form of undulatory movements caudal to the lesion site.
If the synaptic sites left vacant by spinal cord transection
are replaced with excitatory synapses from dorsal root
fibers or indigenous spinal tracts by way of sprouting, an
increase in the discharge pattern should occur with a
resultant increase in the strength of undulatory movements.
Undulatory movements in the present experiment began

65
immediately upon recovery from anesthesia and increased
in strength up to 60 days postoperative after which they
leveled off. If the postoperative increase in the strength
of undulatory movements is compared to the postoperative
increase in synaptic complement on motor horn' cells caudal
to the lesion and the resultant data plotted on a graph
(Fig. 8A, 8B), the curves of the two postoperative phenomena
are highly correlated. Comparison between boutons on cell
bodies and undulatory movements has a correlation coefficient
of r=.930 (P_<. 01) , and comparison between boutons on motor
horn cells primary dendrites and undulatory movements results
in a correlation coefficient of r=.91 (:P<:01) .
It is therefore highly probable from these data
that the synaptic return on motor horn cells by way of
local sprouting is responsible for the increase in the
strength of undulatory activity caudal to the site of lesion.
To further support this hypothesis, retransection at 90 days
had a minimal effect on both synaptic complement and
undulatory movements (Fig. 8A, 8B).

Figure 8 -
A. Comparison between synapse count on motor
horn cell bodies caudal to the lesion and
undulatory strength (r=.93, I^<.01) .
B. Comparison between synapse count on motor
horn cell dendrites caudal to the lesion
and undulatory strength (r=.91, P_<. 01).

NUMBER OF BOUTONS PER 10i<
67
>-
Q
o
un
z
O
â–º-
D
O
0D
£
z
r 3.0
-2.4
â–  18
-1.2
-0.6
DAYS POSTOPERATIVE
o
z
ul
O
>
ac
<
<
at
O
X
»-
o
z
r 3.0
-2.4
-1.8
-1.2
-0.6
DAYS POSTOPERATIVE
B
STRENGTH IN KGAAS OF UNDULATORY STRENGTH IN KGMS OF UNDULATORY
MOVEMENTS ^ MOVEMENTS

CONCLUSION
Regeneration in the shark spinal cord following
spinal cord transection appears to lie somewhere between
the abortive regeneration usually seen in mammals and the
vigorous regeneration and return of function typical of
teleosts. Although anatomical regeneration of nerve
fibers across the site of lesion does occur, the functional
ramifications are negligible and for all practical purposes
the shark remains paralyzed caudal to the lesion when
attempting to swim. The poor anatomical regeneration and
lack of functional return in sharks is surprising. A
general rule of thumb which is well documented is that
the lower on the phylogenetic scale, the more vigorous
and complete is the central nervous system regenerative
process. The reasoning for this is that more primitive animals
reportedly possess more undifferentiated, pleuri-potential
cells capable of differentiation into neural elements. The
elasmobranchs occupy the third rung up on the vertebrate
phylogenetic ladder immediately below the teleosts, yet show
68

69
a postoperative recovery following spinal cord transection
more akin to mammals than fish.
Perhaps the most significant and interesting
result of this experiment is the strong indication of a
functional correlation between the return of synapses on
motor horn cells caudal to the lesion and the increase in
undulatory strength during the postoperative period.
Although functional correlates have been suggested for the
phenomenon of sprouting in the mammalian central nervous
system (McCouch et al., 1955; Schneider, 1970) the evidence
presented in this experiment is perhaps the most conclusive
to date of a functional correlate to sprouting in the
vertebrate central nervous system.

LITERATURE CITED
Adams, J. H., P. M. Daniel and M. M. L. Prichard 1968
Regrowth of nerve fibers in the neurohypophysis:
Regeneration of a tract of the central nervous system.
J. Physiol. (London), 198:4P-5P.
Adams, J. H., P. M. Daniel and M. M. L. Prichard 1969
Degeneration and regeneration of hypothalamic nerve
fibers in the neurohypophysis after pituitary stalk
section in the ferret. J. Comp. Neurol., 135:121-144.
Adams, J. H., P. M. Daniel and M. M. L. Prichard 1971
Changes in the hypothalamus associated with regeneration
of the hypothalamo-neurohypophysial tract after
pituitary stalk section in the ferret. J. Comp. Neurol.,
142:109-124.
Aronson, L. R. 1963 The central nervous system of sharks
and bony fishes with special reference to sensory and
integrative mechanisms. In Sharks and Survival (Ed.
Perry W. Gilbert) pp. 165-241, D. C. Heath and Co.,
Boston, Massachusetts.
Attardi, D. G. and R. Sperry 1963 Preferential selection
of central pathways by regenerating optic fibers.
Exp. Neurol., 7:46-64.
Bernstein, J. J. 1964 Relation of spinal cord regeneration
to age in adult goldfish. Exp. Neurol., 9:161-174.
Bernstein, J. J. 1970 Anatomy and Physiology of the central
nervous system. In Fish Physiology Vol. IV (Ed. W. S.
Hoar and D. J. Randall) pp. 1-90, Academic Press,
New York, London.
Bernstein, J. J. and M. E. Bernstein 1967 Affect of the
glial ependymal scar and teflon arrest on the
regenerative capacity of the goldfish spinal cord.
Exp. Neurol., 19:25-32.

71
Bernstein, J. J. and M. E. Bernstein 1968 Contact inhibition:
A mechanism of abortive regeneration in the goldfish
spinal cord. Anat. Rec. 160:315-316.
Bernstein, J. J. and M. E. Bernstein 1971 Axonal regeneration
and formation of synapses proximal to the site of lesion
following hemisection of the rat spinal cord. Exp.
Neurol., 30:336-351.
Bernstein, M. E. and J. J. Bernstein 1972 Regeneration in
the spinal cord of the monkey. (In press).
Bernstein, J. J. and J. B. Gelderd 1970 Regenerative
capacity of long spinal tracts in the goldfish. Brain
Res., 19:21-26.
Bernstein, J. J. and J. B. Gelderd 1972 Synaptic complement
formation following regeneration of the goldfish spinal
cord. (In manuscript).
Bjorklund, A. R. Katzman, U. Stenevi and K. A. West 1971
Development and growth of axonal sprouts from noradrenaline
and 5-hydroxytryptamine neurones in the cat spinal cord.
Brain Res., 31:21-33.
Bjorklund, A. and U. Stenevi 1971 Growth of central
catecholamine neurones into smooth muscle grafts in
the rat mesencephalon. Brain Res., 31:1-20.
Brown, J. 0. and G. P. McCouch 1947 Abortive regeneration
of the transected spinal cord. J. Comp. Neurol., 87:131-137.
Cattaneo, D. 1923 I fenomeni degenerativi nelle vie visive
in seguito a lesioni del ñervo ottico. Riv. Pat. Nerv.
28:61-118.
Clemente, C. D. 1955 Structural regeneration in the mammalian
central nervous system and the role of neuroglia and
connective tissue. In Regeneration in the Central Nervous
System (Ed. William F. Windle) pp. 147-161, Charles C.
Thomas, Springfield, Illinois.
Clemente, C. D. 1964 Regeneration in the vertebrate central
nervous system. Intern. Rev. Neurobiol., 6:257-301.
Drummond, C. D., Jr. 1954 The influence of piromen on the
regeneration of the spinal cord in adult Triturus
viridescens. Undergrad. Hon. Thesis, Brown Univ.,
. 'I.

72
Ebbesson, Sven 0. E. 1972 New insights into the organization
of the shark brain. Comp. Biochem. and Physiol. 42:121-130.
Ebbesson, Sven O. E. and John S. Ramsey 1968 The optic
tracts of two species of sharks (Galeocerdo cuvier and
Ginglymostoma cirratum). Brain Res. 8:36-53.
Ebbesson, Sven 0. E. and Dolores M. Schroeder 1971
Connections of the nurse shark's telencephalon. Science
173:254-256.
Freeman, L. W. 1955 Functional recovery in spinal rats.
In Regeneration in the Central Nervous System (Ed.
William F. Windle) pp. 195-207, Charles C. Thomas,
Springfield, Illinois.
Fridberg, G., R. S. Nishioka and W. R. Fleming 1966
Regeneration of the caudal neurosecretory system in
the cichlid teleost Tilapia mossambica. J. Exp.
Zool. 162:311-336.
Gaze, R. M. 1970 The Formation of Nerve Connections.
Academic Press, London and New York.
Gaze, R. M. and M. Jacobson 1963 The path from the retina
to the ipsilateral optic tectum of the frog. J.
Physiol. 165:73-74.
Gaze, R. M. and M. J. Keating 1969 The depth distribution
of visual units in the tectum of the frog following
regeneration of the optic nerve. J. Physiol. 200:128-129.
Gaze, R. M. and M. J. Keating 1970a Further studies on the
restoration of the contralateral retinotectal projection
following regeneration of the optic nerve in the frog.
Brain Res., (In Press).
Gaze, R. M. and M. J. Keating 1970b Regenerated visual
units in the frog. Brain Res., (In Press).
Goldstein, L. 1967 Urea Biosynthesis in Elasmobranchs.
In Sharks, Skates and Rays (Ed. Perry W. Gilbert,
Robert F. Mathewson and David P. Rail) John Hopkins
Press, Baltimore, Maryland.
Goldstein, L., W. W. Oppelt and T. H. Maren 1968 Osmotic
regulation and urea metabolism in the lemon shark
Neqaprion brevirostris. Am. J. Physiol. 215:1493,1497.

73
Goodman, D. C. and J. A. Horel 1966 Sprouting of optic
tract projections in the brain stem of the rat.
J. Comp. Neurol., 127:71-78.
Gray, J. 1936 Studies in animal locomotion. IV. The
neuromuscular mechanism of swimming in the eel.
J. Exp. Biol., 13:170.
Gray, J. and A. Sand 1936a The locomotory rhythm of the
dogfish (Scyllium canicula). J. Exp. Biol. 13:200-209.
Gray, J. and A. Sand 1936b Spinal reflexes of the dogfish
(Scyllium canicula). J. Exp. Biol. 13:210-217.
Guth, L. and W. F. Windle 1970 The enigma of central nervous
regeneration. Exp. Neurol. 28:1-43, Supp. 5.
Hamburger, V. 1955 Regeneration in the central nervous
system of reptiles and of birds. In Regeneration in
the Central Nervous System (Ed. William F. Windle)
pp. 47-53, Charles C. Thomas, Springfield, Illinois.
Harvey, J. E. and H. H. Srebnik 1967 Locomotor activity
and axon regeneration following spinal cord compression
in rats treated with L-thyroxine. J. Neuropath. Exp.
Neurology 26:661-668.
Healey, E. G. 1957 The Nervous System. In The Physiology
Of Fish Vol. 2 (Ed. Margaret E. Brown) pp. 1-119,
Academic Press Inc., New York.
Healey, E. G. 1962 Experimental evidence for regeneration
following spinal section in the minnow (Phoxinus phoxinus).
Nature 194:395-396.
Hibbard, E. 1963 Regeneration of the severed spinal cord
of chordate larvae of Petromyzon marinus. Exp. Neurol.,
7:175-185.
Hooker, D. 1925 Studies on regeneration in the spinal cord.
III. Reestablishment of anatomical and physiological
continuity after transection in frog tadpoles. J.
Comp. Neurol. 38:315-347.
Hooker, D. 1930 Physiological reactions of goldfish with
severed spinal cord. Proc. Soc. Exp. Biol. Med.,
28:89-90.

74
Hooker, D. 1932 Spinal cord regeneration in the young
rainbow fish Lebistes Reticulatus. J. Comp. Neurol.,
56:277-295.
Horder, T. J. 1971 Retention by fish optic nerve fibers
regenerating to new terminal sites in the tectum of
"chemospecific" affinity for their original sites.
J. Physiol. (London) 216:53-55P.
Jacobson, M. and R. M. Gaze 1965 Selection of appropriate
tectal connections by regenerating optic nerve fibers
in adult goldfish. Exp. Neurol., 13:418-430.
Kappers, Ariens C. U. 1906 The structure of the teleostian
and selachian Brain. J. Comp. Neurol, and Psychology.
16:1-110.
Kappers, Ariens C. U., G. C. Huber and E. C. Crosby 1936
The Comparative Anatomy of the Nervous System of
Vertebrates Including Man. Vol. 1, The Macmillan Co.,
New York.
Katzman, R., A. Bjorklund, Ch. Owman, U. Stenevi and K. A.
West 1971 Evidence for regenerative axon sprouting
of central catecholamine neurons in the rat mesencephalon
following electrolytic lesions. Brain Res., 25:579-596.
Keil, J. H. 1940 Functional spinal cord regeneration in
adult Rainbow fish. Soc. Exp. Biol. Med., 43:175-177.
Kirsche, W. 1950 Die regenerativen Vorgange am Ruckenmark
eruachsever Teleostier nach operativer Kontinuitat.
Strennung. Z. Mikroskop. Anat. Forsch. 56:190-265.
Kirsche, W. 1956 Experimentelle Untersuchunger uber die
regeneration des durchtrennten Ruckenmarkes von
Amblystoma mexacanum. Z. Mikroskop. Anat. Forsch.
65:512-586.
Kirsche, W. 1965 Regenerative Vorgange im Gehirn und
Ruckenmark. Ergeb. Anat. Entiv-gesch. 38:143-194.
Koppanyi, T. 1955 Regeneration in the central nervous
system of fishes. In Regeneration in the Central
Nervous System (Ed. William F. Windle) pp.3-19,
Charles C. Thomas, Springfield, Illinois.

75
Koppanyi, T. and P. Weiss 1922 Funktionelle regeneration
des Ruckenmarkes bei anamniern. Anz. Akad. Wiss. Wien.
Math. Naturre Kl. 59:206-219.
Levi-Montalcini, R. and B. Brooks 1960 Excessive growth
of the sympathetic ganglis evoked by a protein isolated
from the mouse salivary gland. Proc. Nat. Acad. Sci.
(Wash.) 46:373-384.
Lissmann, H. W. 1946a The neurological basis of the
locomotory rhythm in the spinal dogfish (Scylliuro
canícula, Acanthius vulgaris) I. Reflex Behavior.
J. Exp. Biol. 23:143-161.
Lissmann, H. W. 1946b The neurological basis of the
locomotory rhythm in the spinal dogfish (Scyllium
canícula, Acanthius vulgaris) II. The effect of
deafferentation. J. Exp. Biol. 23:162-176.
Littrell, J. L. 1955 Apparent functional restitution in
piromen treated spinal cats In Regeneration in the
Central Nervous System (Ed. William F. Windle)
pp. 219-228, Charles C. Thomas, Springfield, Illinois.
Littrell, J. L., D. Bunnell, W. F. Agnew, J. O. Smart and
W. F . Windle 1953 Effects of a bacterial pyrogen on
hind-limb function in spinal cats. Anat. Rec., 115:430-436.
Liu, C. N. and W. W. Chambers 1958 Intraspinal sprouting of
dorsal root axons. Arch. Neurol. 79:46-61.
Lund, R. D. and J. S. Lund 1971 Synaptic adjustment after
deafferentation of the superior colliculus of the rat.
Science, 171:804-807.
Marón, K. 1959 Regeneration capacity of the spinal cord
in Lampetra fluviatilis larvae. Folia Biol., 7:179-189.
McCouch, G. P., G. M. Austin and C. Y. Liu 1955 Sprouting
of new terminals as a cause of spasticity. Am. J.
Physiol. 183:642.
Moore, R. Y., A. Bjorklund and U. Stenevi 1971 Plastic
changes in the adrenergic innervation of the rat
septal area in response to denervation. Brain Res.
(In Press).

76
Niazi, I. A. 1963 The histology of tail regeneration in
the animocoetes. Can. J. Zool., 41:125-145.
Nicholas, J. S. and D. Hooker 1928 Progressive cord
degeneration and collateral transmission of spinal
impulses following section of the cord in albino
rat fetuses. Anat. Rec. 38:24-32.
Nieuwenhuys, R. 1964 Comparative anatomy of the spinal
cord. Prog. Brain Res. 11:1-57.
Pearcy, J. F. and T. Koppanyi 1924 A further note on
regeneration of the cut spinal cord in fish. Proc.
Soc. Exp. Biol. Med., 22:17-19.
Piatt, J. 1955a Regeneration in the central nervous
system of amphibia. In Regeneration in the Central
Nervous System (Ed. William F. Windle) pp. 20-46,
Charles C. Thomas, Springfield, Illinois.
Piatt, J. 1955b Regeneration of the spinal cord in the
salamander. J. Exp. Zool., 129:177-207.
Piatt, J. and M. Piatt 1958 Transection of the spinal
cord in the adult frog. Anat. Rec. 131:81-95.
Raisman, G. 1966 The connexions of the septum. Brain
89:317-348.
Raisman, G. 1969a A comparison of the mode of termination
of the hippocampal and hypothalamic afferents to the
septal nuclei as revealed by the electron microscopy
of degeneration. Exp. Brain Res. 7:317-343.
Raisman, G. 1969b Neuronal plasticity of the septal nuclei
of the adult rat. Brain Res. 14:25-48.
Ramon y Cajal, S. 1928 Degeneration and Regeneration of the
Nervous System. 1:47-51.
Ramon y Cajal, S. 1960 Studies on Vertebrate Neurogenesis,
L. Guth, Translator, Charles C. Thomas, Springfield,
Illinois.

77
Rasmussen, G. L. 1957 Selective silver impregnation of
synaptic endings In New Research Techniques of
Neuroanatomy, (Ed. W. F. Windle) pp. 27-39, Charles
C. Thomas, Springfield, Illinois.
Rasmussen, L. E. 1971 Organ distribution of exogenous
â– ^C-urea in elasmobranchs with special regard to the
nervous system. Comp. Biochem. Physiol. 40A:145-154.
Rasmussen. L. E. and R. A. Rasmussen 1967 Comparative
protein and enzyme profiles of the cerebrospinal fluid,
extradural fluid, nervous tissue and sera of
elasmobranchs In Sharks, Skates and Rays (Ed. Perry W.
Gilbert, Robert F. Mathewson and David P. Rail)
pp. 361-380, John Hopkins Press, Baltimore, Maryland.
Schneider, G. E. 1970 Mechanisms of functional recovery
following lesions of visual cortex and superior
colliculus in neonate and adult hamsters. Brain,
Behavior and Evol. 3:295-323.
Scott, D., Jr., E. Gutmann and P. Horsky 1966 Regeneration
in spinal neurons: Proteosynthesis following nerve
growth factor administration. Science 152:787-788.
Scott, D. and C. N. Liu 1964 Factors promoting regeneration
of spinal neurons: Positive influence of nerve growth
factor. Prog, in Brain Res. 13:127-150.
Segaar, J. 1965 Behavioral aspects of degeneration and
regeneration in fish brain: A comparison with higher
vertebrates. Prog. Brain Res. 14:143-231.
Sharma, S. C. 1972 Reformation of retinotectal projections
after various tectal ablations in goldfish. Exp.
Neurol. 34:171-182.
Smith, H. 1929 The composition of the body fluids in
elasmobranchs. J. Biol. Chem. 81:407-419.
Sperry, R. W. 1944 Optic nerve regeneration with return
of vision in anurans. J. Neurophysiol. 7:57-70.
Sugar, 0. and R. W. Gerard 1940 Spinal cord regeneration
in the rat. J. Neurophysiol. 3:1-19.

78
Ten Cate, J. and Ten Cate-Kazejawa 1933 La coordination des
mouvements locomoteurs apres la section transversale de
la moelle epiniere chez les requins. Arch. Neerl.
Physiol. 18:15-23.
Von Lenhossek, M. 1892 Beobachtungen an den spinalganglien
und deni Ruchenmark von Pristiurusembryonen. Anat.
Anz. Bd. 7:519.
Von Lenhossek, M. 1895 Der fienere bau des nervensystems
im lechte neuester Forschungen. Aufl. Fischer, Berlin.
Windle, W. F. 1955 Comments on regeneration in the human
central nervous system. In Regeneration in the Central
Nervous System (Ed. William F. Windle) pp. 265-272,
Charles C. Thomas, Springfield, Illinois.
Windle, W. F. 1956 Regeneration of axons in the vertebrate
central nervous system. Physiol. Rev. 36:427-440.
Windle, W. F. and W. W. Chambers 1950a Regeneration in the
spinal cord of the cat and dog. J. Comp. Neurol.
93:241-257.
Windle, W. F. and W. W. Chambers 1950b Spinal cord
regeneration associated with a cellular reaction induced
by administration of a purified bacterial pyrogen.
Abst. V. Internat. Anat. Comg., Oxford, p. 196.
Windle, W. F. and W. W. Chambers 1951 Regeneration in the
spinal cord of the cat and dog. Arch. Neurol. Psychiat.,
Chi., 65:261-262.
Windle, W. F., C. D. Clemente and W. W. Chambers 1952
Inhibition of formation of a glial barrier as a means
of permitting peripheral nerve to grow into the brain.
J. Comp. Neurol. 96:359-370.
Yoon, M. 1971 Reorganization of retinotectal projection
following surgical operations on the optic tectum in
goldfish. Exp. Neurol. 33:395-411.

BIOGRAPHICAL SKETCH
John Bruce.Gelderd was born during a German
bombing raid in Wasquehal, France on September 21, 1939.
He attended grade school in Broadview, Illinois, and Clearwater,
Florida, and graduated from Clearwater High School in 1957.
After a 4^ year stint in the U. S. Air Force, he attended
St. Petersburg Jr. College from September, 1962, to May,
1964. He entered the University of Florida in September,
1964 and received a bachelor's degree in zoology in 1967.
He began his graduate studies toward the Doctor of
Philosophy degree at the University of Florida in September,
1968.
He has accepted a faculty position in the
Department of Anatomy at Louisiana State University in
New Orleans, Louisiana.
79

I certify that I have read this study and that in my
opinion it conforms to acceptable standards of scholarly
presentation and is fully adequate, in scope and quality,
as a dissertation for the degree of Doctor of Philosophy.
t.
ÍíIülL
Bernstein, Chairman
iate Professor of Neuroscience
I certify that I have read this study and that in my
opinion it conforms to acceptable standards of scholarly
presentation and is fully adequate, in scope and quality,
as a dissertation for the degree of Doctor of Philosophy.
A
F. A. King, Chairman of
Department of Neuroscience
I certify that I have read this study and that in my
opinion it conforms to acceptable standards of scholarly
presentation and is fully adequate, in scope and quality
as a dissertation for the degree of Doctor of Philosophy.
C./JJ. Vierck,
Associate Processor of Neuroscience
I certify that I have read this study and that in my
opinion it conforms to acceptable standards of scholarly
presentation and is fully adequate, in scope and quality,
as a dissertation for the degree of Doctor of.Philosophy.
' / f k
John B. Munson
Assistant Professor, Neuroscience

This dissertation was submitted to the Department of
Neuroscience in the College of Medicine and to the Graduate
Council, and was accepted as partial fulfillment of the
Dean, Graduate School

04
UNIVERSITY OF FLORIDA
3 1262 08554 3782




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