An electron microscopic analysis of the left phrenic nerve in the rat

In this electron microscopic study, the axonal categories in the left phrenic nerve at its entrance to the diaphragm have been determined. At a level 3 mm rostral to the diaphragm, the left phrenic nerve contains approximately 700 axons: 57% are myelinated and 43% are unmyelinated. The dorsal root ganglion cells give rise to 31% of the myelinated axons and the ventral root contributes 69%. Of the unmyelinated axons, the dorsal root ganglion cell contributes 59%, the cervical sympathetic chain 24%, and 17% course through the ventral roots. These ventral root unmyelinated axons are presumably preganglionic efferents since the proximal stump of the ventral root showed no decrease in unmyelinated axons after ventral rhizotomy.     

Regrowth of axons in lesioned adult rat spinal cord: promotion by implants of cultured Schwann cells

Summary Highly purified populations of Schwann cells were grafted into lesioned adult rat spinal cord to determine if they promote axonal regeneration. Dorsal spinal cord lesions were created by a photochemical lesioning technique. Schwann cells derived from E16 rat dorsal root ganglia, either elongated and associated with their extracellular matrix or dissociated and without matrix, were rolled in polymerized collagen to form an implant 4–6 mm long which was grafted at 5 or 28 days after lesioning. No immunosuppression was used. Acellular collagen rolls served as controls. At 14, 28 and 90 days and 4 and 6 months after grafting, animals were analysed histologically with silver and Toluidine Blue stains and EM. The grafts often filled the lesion and the host borders they apposed exhibited only limited astrogliosis. By 14 days, bundles of unmyelinated and occasional thinly myelinated axons populated the periphery of Schwann cell implants. By 28 days and thereafter, numerous unmyelinated and myelinated axons were present in most grafts. Silver staining revealed sprouted axons at the implant border at 28 days and long bundles of axons within the implant at 90 days. Photographs of entire 1 m plastic cross-sections of nine grafted areas were assembled into montages to count the number of myelinated axons at the graft midpoint; the number of myelinated axons ranged from 517–3214. Electron microscopy of implants showed typical Schwann cell ensheathment and myelination, increased myelin thickness by 90 days, and a preponderance of unmyelinated over myelinated axons. Random EM sampling of five Schwann cell grafts snowed that the ratio of unmyelinated to myelinated axons was highest (201) at 28 days. These ratios implied that axons numbered in the thousands at the graft midpoint. Dissociated Schwann cells without matrix promoted axonal ingrowth and longitudinal orientation as effectively as did elongated Schwann cells accompanied by matrix. There was a suggestion that axonal ingrowth was at least as successful, if not more so, when the delay between lesioning and grafting was 28 rather than 5 days. Acellular collagen grafts did not contain axons at 28 days, the only interval assessed. In sum, grafts of Schwann cells in a rolled collagen layer filled the lesion and were well tolerated by the host. The Schwann cells stimulated rapid and abundant growth of axons into grafts and they ensheathed and myelinated these axons in the normal manner.     

Redistribution of Schwann cells at the developing PNS–CNS borderline. An ultrastructural and autoradiographic study on the S1 dorsal root of the cat

In order to test our hypothesis that myelin-forming Schwann cells early during development, after having been eliminated from their parent axons, colonize neighbouring unmyelinated axons, we studied the distribution of Schwann cells at the PNS–CNS border in the feline S1 dorsal spinal root during pre- and postnatal development using electron microscopy and autoradiography. Myelination of axons peripheral to the PNS–CNS border began about 1.5 weeks before birth. The adult distribution of one-third myelinated and two-thirds unmyelinated axons was noted 3 weeks after birth. Analysis based on to-scale reconstructions of axon and Schwann cell samples from the first 6 postnatal weeks gave the following results. 1) CNS tissue appeared in the proximal part of the root around birth and expanded peripherally during the first three postnatal weeks. (2) The number of Schwann cells associated with myelinated axons decreased. (3) The number of Schwann cells associated with unmyelinated axons increased. (4) The mitotic activity of the Schwann cells was low at birth and nil after the first postnatal weak. (5) Apoptotic cell units were virtually absent. (6) Aberrant Schwann cells, i.e. short and very short Schwann cells with distorted and degenerating myelin sheaths, were common. (7) The endoneurial space contained numerous Schwannoid cells i.e. solitary cells surrounded by a basal lamina. (8) Cytoplasmic contacts between unmyelinated axons and aberrant Schwann cells or Schwannoid cells were observed. We take these results to support our hypothesis.     

In vivo proliferation,migration and phenotypic changes of Schwann cells in the presence of myelinated fibers

Following injury to a peripheral nerve, changes in the behavior of Schwann cells help to define the subsequent microenvironment for regeneration. Such changes, however, have almost exclusively been considered in the context of Wallerian degeneration distal to an injury, where loss of axonal contact or input is thought to be critical to the changes that occur. This supposition, however, may be incorrect in the proximal stumps where axons are still in contact with their cell bodies. In this work, we studied aspects of in vivo Schwann cell behavior after injury within the microenvironment of proximal stumps of transected rat sciatic nerves, where axons are preserved. In particular we studied this microenvironment proximal to the outgrowth zone, in an area containing intact myelinated fibers and a perineurial layer, by using double immunolabelling of Schwann cell markers and 5-bromo-2'-deoxyuridine (BrdU) labeling of proliferating cells.In normal sciatic nerve, Schwann cells were differentiated, in an orderly fashion, into those associated with unmyelinated fibers that labeled with glial fibrillary acidic protein (GFAP) and those associated with myelinated fibers that could be identified by individual axons and myelin sheaths. After sciatic nerve transection, there was rapid and early expansion in the population of GFAP-labeled cells in proximal stumps that was generated in part, by de novo expression of GFAP in Schwann cells of myelinated fibers. Schwann cells from this population also underwent proliferation, indicated by progressive rises in BrdU and GFAP double labeling. Finally, this Schwann cell pool also developed the property of migration, traveling to the distal outgrowth zone, but also with lateral penetration into the perineurium and epineurium, while in intimate contact with new axons.The findings suggest that other signals, in the injured proximal nerve stumps, beyond actual loss of axons, induce 'mature' Schwann cells of myelinated axons to dedifferentiate into those that up-regulated their GFAP expression, proliferate and migrate with axons.     

Fibre composition of the ventral roots L7 and S1 in the owl monkey (Aotus trivirgatus)

Summary The ventral roots L7 and S1 of the owl monkeyAotus trivirgatus, were examined by electron microscopy. On average, these roots contain 2950 and 1837 myelinated axons respectively. In both roots the myelinated axons have bimodal size distributions, but the S1 root contains more small myelinated axons. Both roots contain a substantial proportion of unmyelinated axon profiles (UAP). In the L7 root the proportion of UAP decreases as the spinal cord is approached, from 19% distally to 5% in the juxtamedullary rootlets. Unmyelinated and very small myelinated CNS-type axons have not been observed in the L7 transitional region. The average S1 root contains some 40% unmyelinated axons at all examined proximo-distal levels. Unmyelinated/ very small myelinated axons are easily found on the CNS side of the S1 transitional region, in direct relation to motoraxon bundles. Bundles of unmyelinated and small myelinated axons occur in the ventral pia mater of both segments. The unmyelinated axons in the L7 root of the owl monkey appear to be arranged like those in the feline L7 ventral root, possibly representing afferents. It is likely that most unmyelinated and small myelinated axons in the ventral root S1 are autonomic efferents.     

Defective myelination in the optic nerve of the Browman-Wyse (BW) mutant rat

Summary The Browman-Wyse (BW) rat displays a spectrum of ocular abnormalities which include myelination by Schwann cells of retinal ganglion cell (RGC) axons within the retina. Immunohistochemical and ultrastructural studies of the optic nerves of adult BW rats (30–60 days of age) with myelinated intraretinal axons were performed. Although individual nerves displayed considerable morphological variability, all were characterized by an initial dysmyelinated proximal segment which was separated from a normally myelinated distal segment by a transitional junctional zone. The proximal segment contained axons which were predominantly unmyelinated: where myelination occurred, almost all sheaths were P_o-positive, proteolipid protein-negative, and the myelinating cell was a Schwann cell. In the distal segment the distribution of myelinated axons appeared to be normal, sheaths were PLP⁺, and the myelinating cell was an oligodendrocyte. Within the proximal segment, axons that were myelinated by Schwann cells were isolated by a basal lamina and expanded extracellular spaces from the bulk of other RGC axons within the optic nerve. Few carbonic anhydrase (CAII)⁺ or GalC⁺ oligodendrocytes were seen in proximal segments that contained Schwann cells: anti-CAII antibody stained atypical cells within the proximal segments which did not resemble CAII⁺ oligodendrocytes in the distal segment, and which were probably GalC^–. Astrocytes appeared normal throughout the length of the nerve, and there was no morphological specialization at the junctional zone similar to that at the lamina cribrosa. The possible source (s) of the intraneural Schwann cells, and the pathogenetic mechanisms underlying the aberrant myelination of RGC axons within the BW optic nerve are discussed.     

Vagal nerve maturation in the fetal lamb: An ultrastructural and morphometric study

The maturation of the left vagal nerve was studied in the fetal lamb by transmission electron microscopy and by computer-assisted morphometry of sections of the entire nerve at seven gestational ages between 79 and 145 days (term is 147 days) and in the adult ewe. The number of unmyelinated axons per Schwann cell progressively decreased from 25 to 55 at 79 days to 1 to 5 at near-term. Unmyelinated axons of various sizes were enclosed within a single Schwann cell at all ages, but the mean axonal diameter increased in inverse relation to the number of unmyelinated axons. A few Schwann cells enclosed two myelinated axons, but in most instances myelination did not begin until a 1:1 ratio was achieved; some single axons with a Schwann cell remained unmyelinated in the adult. Myelinated fibers were rare at 79 days but myelination progressed rapidly thereafter until the adult ratio of myelinated: unmyelinated fibers was reached at about 100 days; myelinated axons were not uniformly distributed. The myelin sheaths and axons of small fibers progressively increased in diameter in late gestation, but new large fibers were not added. Early myelinating fibers and immature unmyelinated axons contained more microtubules than neurofilaments; neurofilaments predominated in mature axons with or without myelin. Cross-linkages between neurofilaments were already evident by 79 days. Maturation of the vagal nerve thus occurs first by an increase in number of myelinated fibers and then by an increase in the size of each fiber in this fixed population. The bimodal distribution in the size histogram of myelinated fibers is not achieved until 134 days gestation and correlates well with physiological maturation of respiratory patterns. © 1993 Wiley-Liss, Inc.     

The fine structure of the Golgi tendon organ

Summary The Golgi tendon organ (GTO) has a capsule composed of cells confluent with the perineural epithelial sheath surrounding the Ib afferent nerve. Fluids of the capsule lumen are isolated from extra-capsular fluids by the capsule wall; its tight-fitting collars form seals through which collagen bundles enter and leave the proximal and distal ends of the fusiform capsule. The capsule lumen between the sealed capsule openings, is divided into longitudinal compartments by delicate processes of septal cells. Collagen bundles spiral down through the longitudinal axis of the GTO. As they descend through the lumen, the bundles are distinctly separated from each other by fluid-filled spaces. These collagen bundles divide, twist, and regroup over short longtiudinal distances and unmyelinated axons interwine among them. Compartments which are densely filled with collagen are not innervated. Axon profiles appear in four forms: (I) myelinated branches of Ib axon, (II) unmyelinated axons surrounded by Schwann cell processes and basal lamina, (III) axons covered only by basal lamina, and (IV) axons with bare surfaces. We postulate that increased tensile forces on the collagen bundles caused by muscle contraction tighten the braided collagen bundles thereby squeezing and distorting the axon terminals. The proposed mechanical events are consistent with known discharge characteristics of Ib axons.     

Sensory fibres in ventral roots L7 and S1 in the cat

1. Receptive fields were determined for ninety-eight unmyelinated and 132 myelinated axons in the L7 and S1 cat ventral roots.2. Seventy of the ninety-eight unmyelinated axons had their receptive fields in somatic structures, the skin and deep tissues.3. Of the seventy unmyelinated axons with somatic receptive fields, thirty-five were mechanical nociceptors, fifteen were mechanical and thermal nociceptors, eleven were deep nociceptors, six were thermal receptors, and three were low threshold mechanoceptors.4. Twenty-four of the ninety-eight unmyelinated axons had their receptive fields in visceral structures: the intestine, bladder and vagina.5. We confirm the work of others that myelinated fibres attached to peripheral receptive fields can be found in ventral roots and that the receptive fields and functional qualities of these fibres are as one would expect of dorsal root fibres for the same segments.6. A previous study demonstrated that approximately 30% of the axons in the L7 and S1 cat ventral roots are unmyelinated and arise from dorsal root ganglion cells. The present study confirms that these axons are sensory and that the axons are predominantly cutaneous nociceptors and visceral afferents. Thus it is concluded that the L7 and S1 cat ventral roots have a major sensory component.     

De- and remyelination in spinal roots during normal perinatal development in the cat: a brief summary of structural observations and a conceptual hypothesis

We have studied the perinatal development of large myelinated axons (adult D > 10 microm) in cat ventral and dorsal lumbosacral spinal roots using autoradiography and electron microscopy (serial section analysis). These axons acquire their first myelin sheaths 2-3 weeks before birth and show nearly mature functional properties first at a diameter of 4-5 microm, i.e. 3-4 weeks after birth. The most conspicuous event during this development takes place around birth, when a transient primary myelin sheath degeneration strikes already well myelinated although short 'aberrant' Schwann cells. The aberrant Schwann cells become completely demyelinated, then measuring about 10 microm in length, and are subsequently eliminated from their parent axons. Morphometry indicates that on average 50% of the Schwann cells originally present along a prospective large spinal root axon suffer elimination. Here it should be noted that in cat lumbo-sacral spinal roots, the longitudinal growth of myelinated Schwann cells that belong to the group containing what will be the largest fibers is on average twice that of their parent axons. The elimination phenomenon is particularly striking in the dorsal roots close to the spinal cord where CNS tissue invades the root for several hundred micrometres. Our observations suggest that, once demyelinated and then eliminated, Schwann cells (i.e. aberrant Schwann cells) colonize neighbouring axons, future myelinated as well as future unmyelinated ones. In the former case the immigrant Schwann cells appear to start myelin production, possibly risking a second demyelination and elimination. We take our observations to indicate that Schwann cells in the cat, during normal development, may switch iteratively between a 'myelin-producing' and a 'non-myelin-producing' phenotype. From a functional point of view the transient presence along a myelinated axon of intercalated unmyelinated segments approximately 10 microm long, due to aberrant Schwann cells, would mean a slowing down of the action potential. The rapid disappearance of aberrant Schwann cells during the two first postnatal weeks could then explain the progressing normalization of the leg-length conduction time.     

Regeneration of axons in the optic nerve of the adult Browman-Wyse (BW) mutant rat

Summary We have studied the regeneration of axons in the optic nerves of the BW rat in which both oligodendrocytes and CNS myelin are absent from a variable length of the proximal (retinal) end of the nerve. In the optic nerves of some of these animals, Schwann cells are present. Axons failed to regenerate in the exclusively astrocytic environment of the unmyelinated segment of BW optic nerves but readily regrew in the presence of Schwann cells even across the junctional zone and into the myelin debris filled distal segment. In the latter animals, the essential condition for regeneration was that the lesion was sited in a region of the nerve in which Schwann cells were resident. Regenerating fibres appeared to be sequestered within Schwann cell tubes although fibres traversed the neuropil intervening between the ends of discontinuous bundles of Schwann cell tubes, in both the proximal unmyelinated and myelin debris laden distal segments of the BW optic nerve. Regenerating axons never grew beyond the distal point of termination of the tubes. These observations demonstrate that central myelin is not an absolute requirement for regenerative failure, and that important contributing factors might include inhibition of astrocytes and/or absence of trophic factors. Regeneration presumably occurs in the BW optic nerve because trophic molecules are provided by resident Schwann cells, even in the presence of central myelin, oligodendrocytes and astrocytes. All the above experimental BW animals also have Schwann cells in their retinae which myelinate retinal ganglion cell axons in the fibre layer. Control animals comprised normal Long Evans Hooded rats, BW rats in which both retina and optic nerve were normal, and BW rats with Schwann cells in the retina but with normal, i.e. CNS myelinated, optic nerves. Regeneration was not observed in any of the control groups, demonstrating that, although the presence of Schwann cells in the retina may enhance the survival of retinal ganglion cells after crush, concomitant regrowth of axons cut in the optic nerve does not take place.     

Unmyelinated fibers in the cervical and lumbar ventral roots of the cat

Ventral spinal roots at all spinal levels in humans contain many unmyelinated axons, as do all spinal roots of the cat which have been examined. Rats, however, have fewer unmyelinated fibers in cervical and certain lumbar ventral roots. To determine if unmyelinated fibers are distributed uniformly in ventral roots of the cat, cervical and lumbar ventral roots were examined by light and electron microscopy, and axons were counted. Lesions were made to determine the origin of unmyelinated fibers in ventral root L6. From 5 to 15% of the axons in cervical ventral roots are unmyelinated. This distribution of unmyelinated fibers in the cat is similar to the distribution in humans, although the cervical ventral roots of the cat contain relatively fewer unmyelinated fibers than the cervical ventral roots of humans. Approximately 24% of the axons in ventral root L6 are unmyelinated. More than 90% of these axons degenerate proximal to, but not distal to, a ventral rhizotomy and ipsilateral to a dorsal root ganglionectomy. Thus, most unmyelinated axons in ventral root L6 appear to arise from dorsal root ganglion cells. Much variation in the number and percent of unmyelinated axons in ventral roots at the same spinal level exists between individual cats and between opposite sides of the same cat.     

Different multiple regeneration capacities of motor and sensory axons in peripheral nerve

Abstract After peripheral nerve injury, axons often project sprouts from the node of Ranvier proximal to the damage site. It is well known that one parent axon can sprout and maintain several regenerating axons. If enough endoneurial tubes in the distal stump are present for the regenerating axons to grow along, then the number of mature myelinated nerve fibers in the distal stump will be greater than the number in the proximal stump. "Multiple regeneration" is used to describe this phenomenon in the peripheral nerve. According to previous studies, a prominent nerve containing many axons can be repaired by the multiple regenerating axons sprouting from another nerve that contains fewer axons. Most peripheral nerves contain a mixture of myelinated motor and sensory axons as well as unmyelinated sensory and autonomic axons. In this study, a multiple regeneration animal model was developed by bridging the proximal common peroneal nerve with the distal common peroneal nerve and the tibial nerve. Differences in the multiple regeneration ratio of motor and sensory nerves were evaluated using histomorphometry one month after ablating the dorsal root ganglion (DRGs) and ventral roots, respectively. The results suggest that the motor nerves have a significantly larger multiple regeneration ratio than the sensory nerves at two different time points.     

Regrowth of PNS axons through grafts of the optic nerve of the Browman-Wyse (BW) mutant rat

Summary We have examined the behaviourin vivo of regenerating PNS axons in the presence of grafts of optic nerve taken from the Browman-Wyse mutant rat. Browman-Wyse optic nerves are unusual because a 2–4 mm length of the proximal (retinal) end of the nerve lacks oligodendrocytes and CNS myelin and therefore retinal ganglion cell axons lying within the proximal segment are unmyelinated and ensheathed by processes of astrocyte cytoplasm. Schwann cells may also be present within some proximal segments. Distally, Browman-Wyse optic nerves are morphologically and immunohistochemically indistinguishable from control optic nerves.When we grafted intact Browman-Wyse optic nerves or triplets consisting of proximal, junctional and distal segments of Browman-Wyse optic nerve between the stumps of freshly transected sciatic nerves, we found that regenerating axons avoided all the grafts which did not contain Schwann cells, i.e., proximal segments which contained only astrocytes; regions of Schwann cell-bearing proximal segments which did not contain Schwann cells; junctional and distal segments (which contained astrocytes, oligodendrocytes and CNS myelin debris). However, axons did enter and grow through proximal segments which contained Schwann cells in addition to astrocytes. Schwann cells were seen within grafts even after mitomycin C pretreatment of sciatic proximal nerve stumps had delayed outgrowth of Schwann cells from the host nerves; we therefore conclude that the Schwann cells which became associated with regenerating axons within the grafts of Browman-Wyse optic nerve were derived from an endogenous population. Our findings indicate that astrocytes may be capable of supporting axonal regeneration in the presence of Schwann cells.     

Organization of peripheral nerves and spinal roots of the Atlantic stingray, Dasyatis sabina

1. The sizes and numbers of axons in peripheral nerves and spinal roots were investigated in the stingray, Dasyatis sabina. 2. The axons of the dorsal and ventral roots do not mingle in peripheral nerves of this animal as they do in higher vertebrates. Thus, it was usually possible to split the peripheral nerve into two portions, one containing only dorsal root axons, the other containing only ventral root axons. This feature was useful for the analysis of certain aspects of spinal cord organization. 3. The fact that dorsal and ventral root axons were segregated in peripheral nerves enabled us to demonstrate, without experimental surgery, that the central processes of the dorsal root ganglion cells and the proximal ventral root axons were 10-20% narrower, on the average, than the distal processes of the same dorsal root ganglion cells or the distal parts of the same ventral root axons. 4. The stingray is remarkable in having very few unmyelinated axons in the dorsal roots, ventral roots, or peripheral nerves. This paucity of unmyelinated axons distinguishes the Atlantic stingrays from all other vertebrates whose roots and nerves have been examined for unmyelinated fibers. 5. Similar findings were obtained for one spotted eagle ray (Aetobatus narinari) and two cow-nose rays (Rhinoptera bonasus).     

Occurrence of fasciculated microtubules at nodes of Ranvier in rat spinal roots

Summary Fascicles of microtubules, previously considered to be unique to the initial segments of myelinated axons, were found at nodes of Ranvier of sensory and motor axons in rat spinal roots, though their occurrence was limited to the proximal portion of the axons. No fasciculated microtubules were noticed in the internodes of myelinated axons or in unmyelinated axons. In transverse sections, microtubules in fascicles were characteristically cross-linked by short filamentous strands at a centre-to-centre distance of 36–40 nm. In sensory axons, the density of fasciculated microtubules at the node of Ranvier was 45–48% at the level of the dorsal root ganglion and decreased progressively as thin sections were made more distal to the ganglion in both directions. In motor axons, microtubule-fascicles involved about 24% of the microtubules in the ventral rootlets, but they were rarely seen in distal portions of the ventral roots. No fasciculation of microtubules was discerned in the sciatic and saphenous nerves. These findings suggest that cross-linking protein(s) for fasciculation, which are synthesized in the perikaryon and primarily used for the initial segment, may be further transported to fasciculate microtubules at some proximal nodes of Ranvier.     

Acellular nerve graft re-seeded by Schwann cells migrating from the nerve stump can stimulate spinal motoneurons for functional reinnervation of the rat muscle

The acellular nerve graft was utilised to restore a functional reinnervation of the biceps brachii muscle from the motoneuron pool of the cervical spinal cord. The musculocutaneous nerve stump was sutured to an acellular nerve graft, the opposite end of which was inserted into the cervical spinal cord cranial to the avulsed C5 ventral root. The acellular nerve graft was repopulated by Schwann cells heavily immunostained for NGFr within 90 days. The Schwann cells migrating from the nerve stump reached the spinal cord grey matter, where they stimulated the motoneurons to send axonal sprouts. The functional reinnervation of the biceps brachii muscle was assessed by means of the behavioural (grooming) test and EMG, the presence of myelinated and unmyelinated axons was demonstrated by light and electron microscopy. The axonal reconnection of the musculocutaneous nerve stump was verified by horseradish peroxidase retrograde labelling of the spinal motoneurons. Moreover, the motoneurons on the operated side of the C5 spinal segment displayed increased immunostaining for GAP-43 in comparison to the contralateral side, whereas the pattern of AChE histochemical reaction was similar on both the operated and contralateral side, of the C5 segment 150 days after acellular graft implantation. The regenerated axons bridged a 4-cm long originally acellular nerve graft to reach and reinnervate the biceps brachii muscle. The reinnervation of the neuromuscular junctions was morphologically determined by immunofluorescence for neurofilaments. The number of myelinated axons in the acellular nerve graft was significantly higher than those growing over the cellular graft, but their diameter was smaller. The results of experiments presented here demonstrate functional recovery of the biceps muscle reinnervation through the acellular nerve graft repopulated by migrating Schwann cells. The process of reinnervation by acellular nerve graft is however delayed and worse in comparison with the cellular graft.     

Time-dependent differences in the increase in GAP-43 expression in dorsal root ganglion cells after peripheral axotomy.

Peripheral axotomy of primary afferent neurons results in the up-regulation of the growth-associated phosphoprotein GAP-43, by dorsal root ganglion cells. We have studied the temporal sequence of GAP-43 expression in those dorsal root ganglion neurons with unmyelinated axons (the small dark cells) and in those with myelinated axons (the large light cells) after sciatic nerve section in the adult rat. Immunoreactivity for the RT 97 neurofilament epitope, which is detectable only in large light dorsal root ganglion cells, was used to differentiate the two types of dorsal root ganglion cell. Within two days of a sciatic nerve section the number of GAP-43-immunoreactive profiles in the ipsilateral ganglion had increased five-fold and this increase persisted for 80 days post-section. While 50% of the small numbers of GAP-43-positive cells in control ganglia were RT 97 positive, only 8% of the large number of GAP-43-immunoreactive cells four days post-section, were RT 97 positive. By 14 days the number of RT 97-positive/GAP-43-positive cells had increased to 29%. This was paralleled by an increase in GAP-43 immunoreactivity in large diameter profiles at 14 days. The signals that alter GAP-43 expression in unmyelinated (small, RT 97 -ve) and myelinated (large, RT 97 +ve) afferents after peripheral nerve injury appear to operate with different time-courses.     

Acute implantation of an avulsed lumbosacral ventral root into the rat conus medullaris promotes neuroprotection and graft reinnervation by autonomic and motor neurons

Trauma to the conus medullaris and cauda equina may result in autonomic, sensory, and motor dysfunctions. We have previously developed a rat model of cauda equina injury, where a lumbosacral ventral root avulsion resulted in a progressive and parallel death of motoneurons and preganglionic parasympathetic neurons, which are important for i.e. bladder control. Here, we report that an acute implantation of an avulsed ventral root into the rat conus medullaris protects preganglionic parasympathetic neurons and motoneurons from cell death as well as promotes axonal regeneration into the implanted root at 6 weeks post-implantation. Implantation resulted in survival of 44+/-4% of preganglionic parasympathetic neurons and 44+/-4% of motoneurons compared with 22% of preganglionic parasympathetic neurons and 16% of motoneurons after avulsion alone. Retrograde labeling from the implanted root at 6 weeks showed that 53+/-13% of surviving preganglionic parasympathetic neurons and 64+/-14% of surviving motoneurons reinnervated the graft. Implantation prevented injury-induced atrophy of preganglionic parasympathetic neurons and reduced atrophy of motoneurons. Light and electron microscopic studies of the implanted ventral roots demonstrated a large number of both myelinated axons (79+/-13% of the number of myelinated axons in corresponding control ventral roots) and unmyelinated axons. Although the diameter of myelinated axons in the implanted roots was significantly smaller than that of control roots, the degree of myelination was appropriate for the axonal size, suggesting normal conduction properties. Our results show that preganglionic parasympathetic neurons have the same ability as motoneurons to survive and reinnervate implanted roots, a prerequisite for successful therapeutic strategies for autonomic control in selected patients with acute conus medullaris and cauda equina injuries.