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Excitability properties of median and peroneal motor axons.

Threshold tracking was used to compare excitability properties (stimulus-response curves, strength-duration properties, recovery cycle, and threshold electrotonus) of median motor axons at the wrist and peroneal motor axons at the ankle in 12 healthy subjects. Stimulus-response curves and strength-duration properties were similar, though higher stimulus intensities were required for peroneal axons. However, there were significant differences in the recovery cycle of excitability following a conditioning stimulus and in threshold electrotonus. In the recovery cycle, median axons had significantly greater supernormality and late subnormality. In threshold electrotonus, the initial slow threshold changes in response to subthreshold depolarizing and hyperpolarizing currents (S1) were significantly greater in median axons, and there was also greater accommodation to depolarizing currents (S2) and greater threshold undershoot after depolarization. Similar differences in supernormality and the S1 phase of threshold electrotonus were found between peroneal axons at ankle and knee, suggesting that these properties may be dependent on nerve length. When median motor axons at the wrist were compared with peroneal motor axons at the knee, there were no differences in refractoriness and supernormality and only small differences in S1, but the late subnormality and undershoot were significantly greater in the median axons. These findings suggest that, in addition to any length-dependent differences, peroneal axons have a less prominent slow K(+) conductance. We conclude that the properties of different motor axons are not identical and their responses to injury or disease may therefore differ.

Adult↗

Acute ischemia causes axonal stasis, swelling, attenuation, and secondary demyelination.

Microsphere embolization of rat sciatic nerve capillaries results in a central fascicular ischemic core. Twenty-four hours after microembolization, the pathological alterations along the length of 55 myelinated fibers were reconstructed by computer imaging of 2,000 serial semi-thin epoxy sections of a tissue block that extended from just above and into an ischemic core. From proximal to distal, the typical sequence of pathological alterations was: normal----swollen dark axons + thin myelin or demyelination----attenuated axons----axon cytolysis (46 fibers) or normal axons (9 fibers). Because organelle accumulation and axonal swelling were the earliest and most proximal pathological lesions, we infer that regional hypoxia causes axonal stasis as a primary event. Demyelination was found in fibers showing swollen dark and attenuated axons. These findings suggest that axons are selectively vulnerable to acute ischemia and that, depending on severity, the fibers either undergo axonal degeneration or transitory structural alterations without axonal degeneration, the latter consisting of axonal changes and secondary demyelination.

Animals↗

Axonal elongation as a stochastic walk.

A new formula calculates rates of directed axonal growth (elongation or retraction) using measurements of growth cone movements. By explicitly separating changes in axonal length from other nonelongational growth cone movements, the calculated rates reflect the detailed cellular growth mechanisms more directly than previous growth measures. In addition, the formula produces three distinct parameters of axonal elongation: n, a growth step rate; s, a growth step size; and P, a probability that a growth step leads to axonal elongation. For normal and regenerating individual chick and frog axons in culture, the formula has quantitated the following differences: the axon itself can elongate more rapidly in the chick, and the axon elongates in smaller steps in the chick. The underlying dynamics of growth of regenerating axons are quite similar to normal axons, but, in the short term, regenerating axons elongate in larger steps and at a slower rate. The distribution of these new rate measurements suggests that the elongation of axons can be usefully modelled as a one-dimensional stochastic walk.

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Intrinsic connectivity of the rat subiculum: I. Dendritic morphology and patterns of axonal arborization by pyramidal neurons.

The dendritic and axonal morphology of rat subicular neurons was studied in single cells labeled with Neurobiotin. Electrophysiological classification of cells as intrinsic burst firing or regular spiking neurons was correlated with morphologic patterns and cell locations. Every cell had dendritic branches that reached the outer molecular layer, with most cells having branches that reached the hippocampal fissure. All but two pyramidal cells had axon collaterals that entered the deep white matter (alveus). Branching patterns of apical dendrites varied as a function of the cell's soma location along the fissure-alveus axis of the cell layer. The first major dendritic branch point for most cells occurred at the superficial edge of the cell layer giving deep cells long primary apical dendrites and superficial cells short or absent primary apical dendrites. In contrast, basal dendritic arbors were similar across cells regardless of cell position. Apical and basal dendrites of all cells had numerous spines. Superficial and deep cells also differed in axonal collateralization. Deep cells (mostly intrinsically bursting [IB] class) had one or more ascending axon collaterals that typically remained within the region circumscribed by their apical dendrites. Superficial cells (mostly regular spiking [RS] class) tended to have axon collaterals that reached longer distances in the cell layer. Numerous varicosities and axonal extensions were present on axon collaterals in the cell layer and in the apical dendritic region, suggesting intrinsic connectivity. Axonal varicosities and extensions were found on axons that entered presubiculum, entorhinal cortex or CA1, supporting the notion that these were projection cells. Local collaterals were distinctly thinner than collaterals that would leave the subiculum, suggesting little or no myelin on local collaterals and some myelin on efferent fibers. We conclude that both IB and RS classes of subicular principal cells make synaptic contacts in and apical to the cell layer. Based on the patterns of axonal arborization, we suggest that subiculum has at least a crude columnar and laminar architecture, with ascending collaterals of deep cells forming columns and broader axonal arbors of superficial cells serving to distribute activity across multiple columns.

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Regenerating descending axons preferentially reroute to the gray matter in the presence of a general macrophage/microglial reaction caudal to a spinal transection in adult zebrafish.

We analyzed pathway choices of regenerating, mostly supraspinal, descending axons in the spinal cord of adult zebrafish and the cellular changes in the spinal cord caudal to a lesion site after complete spinal transection. Anterograde tracing (by application of the tracer rostral to the spinal lesion site) showed that significantly more descending axons (74%) regenerated in the spinal gray matter of the caudal spinal cord than would be expected from random growth. Retrograde tracing (by application of the tracer caudal to the spinal lesion site) showed that, rostral to the lesion, most of these axons (80%) extended into the major white matter tracts. Thus, ventral descending tracts often were devoid of labeled axons caudal to a spinal lesion but contained many axons rostral to the lesion in the same animals, indicating a pathway switch of descending axons from the white matter to the gray matter. Ascending axons of spinal neurons were not observed regrowing to the rostral tracer application site; therefore, they most likely did not contribute to the axonal populations analyzed. A macrophage/microglia response within 2 days of spinal cord transection, along with phagocytosis of myelin, was observed caudal to the transection by immunohistochemistry and electron microscopy. Nevertheless, caudal to the lesion, descending tracts in the white matter were filled with myelin debris during the time of axonal regrowth, at least up to 6 weeks postlesion. We suggest that the spontaneous regeneration of axons of supraspinal origin after spinal cord transection in adult zebrafish may be due in part to the axons' ability to negotiate novel pathways in the spinal cord gray matter.

Age Factors↗

Pyramidal cell axons show a local specialization for GABA and 5-HT inputs in monkey and human cerebral cortex.

Various mechanisms are thought to control excitation of pyramidal cells of the cerebral cortex. With immunocytochemical methods, we found that the proximal portions of numerous pyramidal cell axons (Pyr-axons) in the human and monkey neocortex are immunoreactive for the serotonin (5-HT) receptor 5-HT-(1A). With double-labeling experiments and confocal laser microscopy, we found that most (93.4%) of the 5-HT(1A)-immunoreactive Pyr-axons present in layers II and III were innervated by parvalbumin-immunoreactive chandelier cell axon terminals. In addition, Pyr-axons were compartmentalized: 5-HT-(1A) receptors were found proximal to inputs from chandelier cells. Although we found close appositions between GABAergic chandelier cell axon terminals and Pyr-axons, suggesting synaptic connections, we did not observe 5-HT-immunoreactive fibers in close proximity to the Pyr-axons. These results suggested that Pyr-axons are under the influence of 5-HT in a paracrine manner (via 5-HT-(1A) receptors) and, more distally, are under the influence of gamma-aminobutyric acid (GABA) in a synaptic manner (through the axons of chandelier cells). The local axonal specialization might represent a powerful inhibitory mechanism by which the responses of large populations of pyramidal cells can be globally controlled by subcortical serotonin afferents, in addition to local inputs from GABAergic interneurons.

Adult↗

C-fiber (Remak) bundles contain both isolectin B4-binding and calcitonin gene-related peptide-positive axons.

Unmyelinated nerve fibers (Remak bundles) in the rodent sciatic nerve typically contain multiple axons. This study asked whether C-fiber bundles contain axons arising from more than one type of neuron. Most small neurons of the lumbar dorsal root ganglion (DRG) are either glial cell line-derived neurotrophic factor dependent or nerve growth factor dependent, binding either isolectin B4 (IB4) or antibodies to calcitonin gene-related peptide (CGRP), respectively. Injection of IB4-conjugated horseradish peroxidase into a lumbar DRG resulted in intense labeling of IB4 axons, with very low background. Visualized by confocal fluorescence, IB4-binding and CGRP-positive nerve fibers originating from different DRG neurons came together and remained closely parallel over long distances, suggesting that these two types of axon occupy the same Remak bundle. With double-labeling immunogold electron microscopy (EM), we confirmed that IB4 and CGRP axons were distinct and were found together in single Remak bundles. Previous studies indicate that some DRG neurons express both CGRP and IB4 binding. To ensure that our immunogold results were not a consequence of coexpression, we studied large populations of unmyelinated axons by using quantitative single-label EM. Tetramethylbenzidine, a chromogen with strong intrinsic signal amplification of IB4-horseradish peroxidase, labeled as many as 52% of unmyelinated axons in the dorsal root. Concomitantly, 97% of the Remak bundles with more than one axon contained at least one IB4-labeled axon. Probabilistic modeling using binomial distribution functions rejected the hypothesis that IB4 axons segregate into IB4-specific bundles (P < 0.00001). We conclude that most Remak bundle Schwann cells simultaneously support diverse axon types with different growth factor dependences.

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Long-term survival of centrally projecting axons in the optic nerve of the frog following destruction of the retina.

A significant number of unmyelinated axons and their synaptic endings in the frog, Rana pipiens, were found to retain a normal morphology long after separation from their cell bodies. At the end of various survival periods following unilateral removal of the retina, horseradish peroxidase (HRP) was administered to the optic nerve stump by a fiber-filling method. In frogs maintained at 20 degrees C, unmyelinated optic nerve axons conducted HRP from the site of application in the orbit to layers A, C, and E of the contralateral optic tectum, even though their retinas had been removed up to 69 days earlier. Such fiber-filling was absent beyond 19 days in other frogs surviving at 35 degrees C. No labeled fibers were continuous with any intracerebral neurons. The HRP was always localized intraaxonally, and the marked axons and terminals were ultrastructurally normal. Counts of surviving axons from electron micrographs of the optic nerves showed that, at 20 degrees C, more than half of the normal complement of unmyelinated axons disappeared in the first 10 days. All the myelinated axons degenerated during the first 6 weeks survival. However, approximately 55,000 normal-appearing unmyelinated axons (12% of the unmyelinated fiber population) persisted in the optic nerve at 10 weeks following removal of the retina. The survival rate was lower at 35 degrees C. In other frogs, one eye was injected with 3H-leucine to initiate axonal transport into the retinal ganglion cell axons. That eye was removed 48 hours later. Autoradiographic analysis of brain sections of frog surviving an additional 31 to 61 days at 20 degrees C showed strong labeling of the optic tract and layers A, C, and E of the contralateral optic tectum. The absence of displaced ganglion cells that might exist within the optic nerve was verified by other observations. It is hypothesized that the potential shown by frog optic axons for long-term survival in the absence of the cell-body expresses a general property of vertebrate (and invertebrate) axons, rather than a special property of the frog optic nerve.

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Electron microscopic and immunohistochemical evidence that unmyelinated ventral root axons make u-turns or enter the spinal pia mater.

The occurrence of unmyelinated and small myelinated axons and of nerve fibres with a substance P-like immunoreactivity was studied in juxtamedullary L7 ventral root fascicles and surrounding pia mater of kittens and cats. Electron microscopic analysis of thin transverse serial sections from this region in adult cats showed that the number of unmyelinated axon profiles decreases rapidly as the CNS is approached, reaching zero near or at the CNS/PNS border. No unmyelinated axons were found to enter the CNS through this root. At least to some extent, the disappearance of unmyelinated ventral root axons from the juxtamedullary root fascicles was due to presence of intrafascicular axonal hairpin loops and to a shift of axons from ventral root fascicles to the pia mater. It was also found that, in the L7 ventral root, the content of unmyelinated axons in the pia mater is lower in kittens than in cats. Fluorescence microscopic examination of specimens incubated with substance P antiserum showed that some looping axons and ventral root-pia mater axons were substance P immunoreactive. These observations suggest the hypothesis that sensory unmyelinated axons might grow through the L7 ventral root and enter the pia mater during postnatal development. Moreover they show that the occurrence of sensory unmyelinated axons in ventral roots does not necessarily contradict the law of Magendie .

Age Factors↗

Microtubules and caliber of central and peripheral processes of sensory axons.

The microtubular content and caliber of sensory axons were studied in the L7 dorsal root, at the distal pole of the L7 spinal ganglion, and in the sural nerve of cats. Calibers of myelinated axons were symmetrical about the ganglion. In contrast, nonmedullated axons were strikingly different; 80% of the population at the root were smaller than 0.4 micron2, whilst just across the ganglion the same group was less than 20%. The microtubule densities of myelinated axons of the root were 11.8 and 6.1 microtubules/micron2 for 3- and 10 microns diameter axons, respectively. Across the ganglion the densities of myelinated axons of equal sizes were 24.2 and 14.4 microtubules/micron2, respectively. These values represent an approximate ratio of 1:2 between central and peripheral microtubule densities. Microtubule densities for nonmedullated axons also decreased with the increase in the cross-sectional area. The densities of root nonmedullated axons ranged between 90 and 10 microtubules/micron2; these were smaller, usually by a factor of three, than the densities of peripheral axons of a similar size (range: 367-44). Contrasting with the differences observed across the ganglion, the microtubular content and caliber of sensory axons seems to be quite uniform along their peripheral course. This is supported by the similar values found in the juxtaganglionic and sural nerves. It is estimated that an axon that contains 90 microtubules/micron2 has 26.7 mg of tubulin per ml of axoplasm in its assembled form, and 3.0 mg/ml if it contains 10 microtubules/micron2; these values are the practical limits of assembled tubulin in axoplasms.(ABSTRACT TRUNCATED AT 250 WORDS)

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Morphology of retinogeniculate X and Y axon arbors in monocularly enucleated cats.

We examined the terminal arbors of single, physiologically identified retinogeniculate X and Y axons from the remaining retinas of adult cats raised from birth with monocular enucleation. These were compared with arbors of X and Y axons in normally reared cats. We used intra-axonal injections of horseradish peroxidase to label each axon after recording its response properties. While the axons in monocularly enucleated cats exhibited normal response properties, both X and Y axons in these cats had abnormally large terminal arbors. Each of the hypertrophied X arbors appeared to be completely confined to the single geniculate lamina A or A1 appropriate to its eye of origin (i.e., lamina A for the contralateral retina and lamina A1 for the ipsilateral retina). In contrast, in addition to their normal terminations, most of the Y arbors seemed to extend well into laminae normally innervated only by the retina that was removed. Thus most or all of the translaminar sprouting previously reported for monocularly enucleated cats appears to reflect extensions of Y axon arbors. These data, in addition to earlier, analogous data from young kittens and cats reared with monocular lid suture, suggest the following sequelae during postnatal development: the retinogeniculate X arbors mature first and develop exuberant arbors that are later competitively pruned as the Y axons expand their innervation of the lateral geniculate nucleus; monocular lid suture prevents the Y axons from succeeding in this competition, so they fail to establish normal arbors and cannot reduce the exuberant X arbors; monocular enucleation offers a less resistant path in the denervated laminae for the rapidly growing Y arbors from the remaining eye, and the expansion of these arbors there reduces the competitive pressure on the exuberant X arbors. Thus, in monocularly enucleated cats, sprouting is limited to Y axons, either because only they possess the capacity to sprout or because they are in the midst of a period of relatively rapid growth at the time of the neonatal enucleation. The X axon arbors are also abnormally large within their appropriate laminae. This occurs presumably because they are able to maintain their immature exuberance, although we cannot rule out the possibility that they are pruned and later regrow to the final size seen in our experiments.

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Axonal arborization in the developing chick retinotectal system.

The growth and arborization of chicken retinal ganglion cell axons have been investigated by means of an intraaxonally transported fluorescent marker in the developing retinotectal system. The fluorescent dye D282 or diI from the carbocyanine group of dyes is taken up by ganglion cells and labels the axon as well as the axonal growth cones and the terminal arborizations on the tectum. Branching and arborization start in the chick retinotectal system on embryonic day 9 (E9). At this stage retinal axons leave the stratum opticum (SO) and invade the stratum griseum et fibrosum superficiale (SGFS), where arborization takes place. On day E12 several axons were found to arborize in the SGFS. At this stage arbors appear to have small branches with less than 4 branching points. The extension of terminal arbors in the anterior/posterior (A/P) and in the dorsal/ventral (D/V) direction was determined for 50 axonal trees at days E13-14 and for 24 arbors at days E15-16. Few axonal terminals were investigated at day E18. The mean A/P extent of axonal terminal trees increases from 0.23 +/- 0.12 to 0.36 +/- 0.22 mm from E13-14 to E15-16 and seems to stay at this order of magnitude on E18. The mean D/V extent increases from 0.23 +/- 0.17 to 0.30 +/- 0.18 mm in the same embryonic period of development. The number of branching points calculated from the same number of axonal trees increases from 7.50 +/- 2.98 at E13-14 to 11.70 +/- 4.10 at E15-16. This number seems to increase further after day E16 achieving values of about 20 to 25 at E18. This was, however, not quantifiable by the technique used here and represents an approximate value estimated from 6 completely labeled terminal fields at E18. The data presented here suggest that the modeling of the final branching pattern in the chick retinotectal system takes place within a relatively short period of embryonic development. Prior to the beginning of terminal arborization two important events contribute to the formation of a retinotopic projection. One event is the change of the D/V position by a minority of axons lying ectopic in terms of retinotopy. Some axons turn at right angles and change their D/V position. The other event is the appearance of side branches along the A/P axis.

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Ultrastructural identification of synaptic terminals from the axon of type 3 interneurons in the cat lateral geniculate nucleus.

Synaptic terminals from the axons of type 3 neurons in the A-laminae of the cat LGN impregnated with the Golgi gold-toning procedure were examined at light and electron microscopic levels. The axons were identified by their somatic origin, thin diameter, and, in one of these cells, by dense undercoating beneath the axolemma, which is a known characteristic of the axon initial segment. The axon of one of the analyzed cells was profusely branched and extended throughout most of lamina A within the dendritic domains of the cell, and both types of processes were oriented along projection lines in LGN. This suggests that the dendrites and axons of type 3 cells receive inputs and exert effects, of probably inhibitory nature, within restricted retinotopic regions of LGN. The vast majority of the axon terminals of these cells were distributed in series along axonal branches. In one of the type 3 cells, however, a dense cluster of terminals arising from a secondary axonal branch was observed. Ultrastructurally, the analyzed synaptic terminals of the type 3 cells contained flattened or pleomorphic synaptic vesicles, dark mitochondria, and established synapses that appeared to be of symmetrical type when the membranes were perpendicularly cut. On the basis of these characteristics these terminals are classified as F boutons, following Guillery's (Z. Zellforsch. 96:1-38, '69), nomenclature. The postsynaptic elements to the axon terminals were dendrites of small to medium size, which received "en passant" synaptic contacts in extraglomerular regions of the geniculate neuropil by the terminals distributed in series. The axon terminals located in clusters, however, made synapses with dendrites in glomerular regions of the neuropil, where they were not seen postsynaptic to retinal or other types of terminals. This is in contrast to the postsynaptic nature of F2 boutons in the same glomeruli, which have been identified as dendritic appendages of the GABA positive type 3 neurons in the cat LGN (Montero: J. Comp. Neurol. 254:228-245, '86). On the other hand, the axonal F terminals differ from F1 boutons in terms of synaptic relations and ultrastructure, since the latter have been shown to be presynaptic to F2s and somata and to contain crowded populations of flat synaptic vesicles which give them a characteristic dark appearance. Terminals equivalent to F1 boutons have been shown to originate from perigeniculate cells in the rat LGN. From these observations it is suggested that the geniculate GABAergic interneurons support two morphologically and functionally different type of inhibitory terminals synapsing the dendrites of relay cells.(ABSTRACT TRUNCATED AT 400 WORDS)

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Intracellular labeling of neurons in the medial accessory olive of the cat: III. Ultrastructure of axon hillock and initial segment and their GABAergic innervation.

The gamma-aminobutyric acid (GABA) synaptic input of identified axons in the cat inferior olive was studied by use of combination of intracellular labeling with horseradish peroxidase and postembedding gold-immunocytochemistry. With this technique olivary cells were physiologically identified and light microscopically reconstructed, and the horseradish peroxidase reaction product and the immunogold labeling were subsequently simultaneously visualized for electron microscopic investigation with the use of serial ultrathin sections. The axons of cell type I (characterized by dendrites which radiate away from the cell body) originated from the soma, whereas those of type II neurons (characterized by dendritic trees which curve back towards the soma) were derived from a primary dendrite. The axons of olivary neurons stand out by the length of their axon hillock (up to 21 microns) and initial segment (up to 40 microns). The hillock forms various spiny appendages which were located within glomeruli together with dendritic spines of other olivary neurons. Axonal spines of type II neurons were more numerous and complex looking than those of type I. The axonal spines, the shaft of the axon hillock, and the transition between the hillock and initial segment were primarily innervated by GABAergic terminals (65%) but non-GABAergic terminals (35%) were present as well. The terminals apposed to the axons of type I neurons contacted mainly the axonal shafts, whereas most of the terminals adjacent to the axons of type II neurons established synaptic contacts with the axonal spines. The initial segments were largely devoid of synaptic input. Distally, the initial segment acquired a myelin sheath.

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First step of selective motoneuron axonal growth: selective outgrowth at discrete sites in the spinal cord.

Selective axonal growth at a series of choice points along pathways is essential for the establishment of precise motoneuron projections. To reveal some of the molecules responsible for this selective growth of motoneuron axons, this study investigates the phenomenon of why motoneurons extend axons outside the spinal cord, whereas interneurons do not. Axonal growth in the chick embryonic spinal cord at early stages of development was examined immunohistochemically. MAb SC1 staining of serial sections selectively revealed the entire distribution of motoneuron axons in the embryo. In the cervical segments, some motoneurons, called dorsal motoneurons, extended axons outside the cord via the dorsal root entry zone. The axons of both dorsal and ventral motoneurons were distributed only in the anterior half of the sclerotome; inside the cord motoneuron cell bodies and axons were distributed evenly along the anterior-posterior axis. Motoneurons and interneurons extended axons during the same period, and although the growth cones of both were intermixed in the same locations, only motoneuron axons grew out from the cord. The outgrowth of all st 19 motoneurons from the cord through either the dorsal or ventral exit zone strongly suggests that extension of their axons outside the cord is a selective rather than random process.

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Effects of delayed transplantation of cultured Schwann cells on axonal regeneration from central nervous system cholinergic neurons.

The introduction of transplants consisting of cultured Schwann cells and their associated extracellular matrix (Sc/ECM) into a central nervous system (CNS) lesion cavity facilitates axonal regeneration from injured, adult mammalian neurons with subsequent reinnervation of their appropriate target (Kromer and Cornbrooks: Proceedings of the National Academy of Sciences of the United States of America 82:6330-6334, 1985). In the present study, the effects of a delayed transplantation procedure on the time course of this regenerative response were evaluated. For these experiments, bilateral CNS lesions were created between the septum and hippocampus by removing the fimbria-fornix pathway. Lesion cavities received either no transplants, transplants of collagen, or Sc/ECM transplants at the time the lesion was created or 6 days later. When no transplants or transplants of collagen were used, axonal sprouts extended for very short distances into the lesion cavity. These axons were not preferentially associated with the collagen transplants nor maintained at long post-lesion survival times. In animals that received Sc/ECM transplants, the number of sprouting axons and the progression of axonal growth along the transplants was much more extensive than for the collagen transplants. Although more axons were detected in cavities that received transplants immediately after the fimbria-fornix lesion, axonal regeneration along the transplants was similar regardless of whether there was a delay in transplanting the Schwann cells. By using histochemical techniques to identify acetylcholinesterase (AChE), regenerating AChE-positive axons were first detected in the cavity at 3 days post-transplantation, were associated with the Sc/ECM transplants by 5 days, and crossed the cavity within 8 days post-transplantation. Regenerating, neurofilament-positive axons crossed the CNS-Sc/ECM transplant interfaces in association with laminin-positive, glial fibrillary acidic protein-positive cellular pathways. Upon reaching the caudal end of the Sc/ECM transplant, the cholinergic axons abandoned the transplant and oriented directly toward the adjacent hippocampus. Both the simultaneous and delayed transplantation paradigms demonstrated a similar reinnervation pattern of AChE-positive fibers in the hippocampus, but there was a more rapid penetration and more extensive arborization of fibers in animals receiving the delayed transplants. Cholinergic fibers initially invaded the dentate gyrus molecular layer and hilus between 8 and 14 days post-transplantation. By 45 days post-transplantation, AChE-positive axons were detected throughout the dentate gyrus and regio inferior, but few fibers were present in regio superior of the hippocampus.(ABSTRACT TRUNCATED AT 400 WORDS)

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Morphology of single vestibulospinal collaterals in the upper cervical spinal cord of the cat. II. Collaterals originating from axons outside the ventral funiculi.

Recent studies have shown that vestibulospinal axons reach the upper cervical spinal cord of the cat via several different funicular routes. The purpose of this study was to describe the projections of those axons travelling outside the well-recognized pathways in the ventral funiculi. These axons are located in the dorsal columns, dorsolateral funiculi, and lateral funiculi. Collaterals of these axons were stained following extracellular injections of Phaseolus vulgaris leucoagglutinin in the medial and descending vestibular nuclei. The trajectories of individual collaterals were reconstructed from serial histological sections. Collaterals arising from axons in the same funiculus usually had the same characteristic appearance. Axons in the lateral funiculi, ipsilateral or contralateral to their cells of origin, gave rise to collaterals that had a simple structure and usually followed a horizontal trajectory across laminae VII and VIII. The boutons of these collaterals were distributed throughout the mediolateral extent of laminae VI and VII and the dorsal half of lamina VIII. In contrast, axons in the dorsolateral funiculi, ipsilateral or contralateral to their cells of origin, terminated primarily in laminae IV and V. Many collaterals of these axons projected either rostrally or caudally and had a narrow mediolateral distribution. The combined distribution of boutons from collaterals originating from axons in the dorsal columns included the dorsal horn and intermediate zone. Although these collaterals were less common and formed a heterogeneous group, they were easily distinguished from collaterals originating from axons travelling in other funiculi. These results indicate that vestibulospinal axons travelling outside the ventral funiculi comprise several distinct systems. Each system travels by a different funicular route and is distinguished by differences in collateral morphology and termination zones.(ABSTRACT TRUNCATED AT 250 WORDS)

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Lack of Purkinje cell loss in adult rat cerebellum following protracted axotomy: degenerative changes and regenerative attempts of the severed axons.

The cerebellar Purkinje cells, due to their geometrical disposition and their high calbindin content, offer an optimal system in which to test the adequacy of current opinions on axotomy effects. We have, therefore, analyzed with calbindin immunostaining the morphological changes of Purkinje cells from 1 day to 6 months after axonal section in the cerebellar white matter. This method allows us to study the morphological changes in their dendrites, cell bodies, and axons. We have also searched for simultaneous changes in glial cells and vascularization by using cell type-specific markers. In addition, an ultrastructural study of Purkinje cells, 7 days after large electrolytic lesions affecting the white matter and the overlying granular layer, was carried out to determine whether amputation of the recurrent collateral system provokes a fast neuronal death. Neither the Purkinje cells axotomized close to their cell bodies (electrolytic lesions) nor those axotomized in the white matter (cerebellar transection) degenerated. Thus, this study demonstrates that Purkinje cells are extremely resistant to axotomy; those severed in the white matter at distances varying from 100 microns to 3 mm remain alive for as long as 6 months. At all survival times studied, axotomized Purkinje cells exhibited few changes in their somata and dendrites, as well as in their glial microenvironment. The major changes occurred in the axonal compartment. Axonal alterations, namely the presence of torpedoes and hypertrophy of the recurrent collateral system, were early events already noticeable 24 hours after the lesion, although they later differed in their time course and spatial distribution. It is remarkable that the distal segments of the central stumps of the cut axons survived in large numbers without any apparent retraction, with their terminal varicosities apposed to the wall of the wound cavity even 6 months after the lesion. Nevertheless, these segments were thinner than normal Purkinje cell axons (axonal atrophy). Despite this apparent immutability, some regenerative attempts did occur in the severed axons, such as axonal sprouts penetrating the deeper region of the granular layer in zones close to the lesion, presence of arciform axons, and hypertrophy of the recurrent collateral system. However, the Purkinje cell axons did not regenerate, and these neurons remained separated from their targets by a cavity in virtually all cases.

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