Differential synaptic inputs to the cell body and proximal dendrites of preganglionic parasympathetic neurons in the rat conus medullaris.

Preganglionic parasympathetic neurons (PPNs) reside in the intermediolateral (IML) nucleus of the rat lumbosacral spinal cord and contribute to the autonomic control of visceral pelvic organs. PPNs provide the final common pathway for efferent parasympathetic information originating in the spinal cord. We examined the detailed ultrastructure of the type and organization of synaptic inputs to the cell body and proximal dendrites of PPNs in the rat conus medullaris. The PPNs were retrogradely labeled by a systemic administration of the B subunit of cholera toxin conjugated to horseradish peroxidase. We demonstrate four distinct types of synaptic boutons in apposition with PPN somata and proximal dendrites: S-type boutons show clear, spheroid vesicles; F-type boutons show flattened vesicles; dense-cored vesicle-type (DCV-type) boutons show a mixture of clear and dense-cored vesicles; L-type boutons were rare, but large, exhibited clear spheroid vesicles, and were only encountered in apposition with the PPN dendrites in our sample. The membrane surface covered by apposed boutons was markedly higher for the proximal dendrites of PPNs, compared with their somata. The inhibitory synaptic influence was markedly higher over the PPN somata compared with their proximal dendrites, as suggested by the higher proportion of putative inhibitory F-type boutons in apposition with the soma and a higher frequency of S-type boutons per membrane length for the proximal dendrites. Our studies suggest that the synaptic input to PPNs originates from multiple distinct sources and is differentially distributed and integrated over the cell membrane surface.

Reimplantation of avulsed lumbosacral ventral roots in the rat ameliorates injury-induced degeneration of primary afferent axon collaterals in the spinal dorsal columns.

Injuries to the cauda equina/conus medullaris portion of the spinal cord can result in motor, sensory, and autonomic dysfunction, and neuropathic pain. In rats, unilateral avulsion of the motor efferents from the lumbosacral spinal cord results in at-level allodynia, along with a corresponding glial and inflammatory response in the dorsal horn of the spinal cord segments immediately rostral to the lesion. Here, we investigated the fate of intramedullary primary sensory projections following a motor efferent lesion. The lumbosacral (L6 and S1) ventral roots were unilaterally avulsed from the rat spinal cord (VRA; n=9). A second experimental group had the avulsed roots acutely reimplanted into the lateral funiculus (Imp; n=5), as this neural repair strategy is neuroprotective, and promotes the functional reinnervation of peripheral targets. A laminectomy-only group served as controls (Lam; n=7). At 8 weeks post-lesion, immunohistochemical examination showed a 42% reduction (P<0.001) in the number of RT97-positive axons in the ascending tracts of the dorsal funiculus of the L4-5 spinal segment in VRA rats. Evidence for degenerating myelin was also present. Reimplantation of the avulsed roots ameliorated axon and myelin degeneration. Axons in the descending dorsal corticospinal tract were unaffected in all groups, suggesting a specificity of this lesion for spinal primary sensory afferents. These results show for the first time that a lesion restricted to motor roots can induce the degeneration of intramedullary sensory afferents. Importantly, reimplantation of the lesioned motor roots ameliorated sensory axon degeneration. These data further support the therapeutic potential for reimplantation of avulsed ventral roots following trauma to the cauda equina/conus medullaris.

Glial reactions and degeneration of myelinated processes in spinal cord gray matter in chronic experimental autoimmune encephalomyelitis.

Multiple sclerosis and experimental autoimmune encephalomyelitis (EAE) result in inflammatory white matter lesions in the CNS. However, information is sparse with regard to the effects of autoimmune demyelinating disease on gray matter regions. Therefore, we studied the late effects of chronic EAE in C57BL/6 mice on the spinal cord gray matter using immunohistochemistry. Here, EAE induced marked astrocytic, microglial, and macrophage activation in the ventral horn gray matter, without any motoneuron loss. Activated caspase-3 was also increased in the ventral horn gray matter. Furthermore, activated poly (ADP-ribose) polymerase (PARP), another apoptotic marker, co-localized with myelin basic protein (MBP) of oligodendrocyte processes, but not with the oligodendroglial cell body marker, adenomatous polyposis coli gene clone CC1 (APC-CC1), or with neurofilament marker (RT-97) or synaptophysin of axonal arbors. However, there was no associated increase in the number of terminal deoxynucleotidyl transferase (TdT) mediated-dUTP nick end labeling positive nuclei in the spinal cord gray matter of EAE mice. In addition, co-localization of MBP and the low-affinity neurotrophin receptor, p75, was demonstrated, further supporting the notion of apoptotic oligodendrocyte process degeneration in the gray matter of EAE mice.

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.

Regenerating supernumerary axons are cholinergic and emerge from both autonomic and motor neurons in the rat spinal cord.

Multipolar neurons in the mammalian nervous system normally exhibit one axon and several dendrites. However, in response to an axonal injury, adult motoneurons may regenerate supernumerary axons. Supernumerary axons emerge from the cell body or dendritic trees in addition to the stem motor axon. It is not known whether these regenerating axons contain neurotransmitters for synaptic transmission at their terminals. Here, using immunohistochemistry for choline acetyltransferase, an enzyme that synthesizes acetylcholine, we demonstrate the emergence of cholinergic supernumerary axons at 6 weeks after a unilateral L5-S2 ventral root avulsion and acute implantation of the avulsed L6 ventral root into the adult rat spinal cord. Light microscopic serial reconstruction of choline acetyltransferase immunoreactive arbors shows that these supernumerary axons originate from both autonomic and motor neurons. The supernumerary axons emerge from the cell body or dendrites, exhibit an abnormal projection pattern within the intramedullary gray and white matters, make frequent abrupt turns in direction, and form bouton-like swellings as well as growth cone-like terminals. Double labeling immunohistochemistry studies show that the choline acetyltransferase immunoreactive supernumerary axons co-localized with two proteins associated with axonal growth and elongation, growth-associated protein 43 and p75, the low affinity neurotrophic factor receptor. Our findings suggest that regenerating supernumerary axons selectively transport and store choline acetyltransferase, supporting the notion that supernumerary axons may develop functional and active synaptic transmission. Therefore, regenerating supernumerary axons may contribute to the plasticity in neural circuits following injury in the adult nervous system.

Quantitative analyses of intracellularly characterized and labeled thalamocortical projection neurons in the ventrobasal complex of primates.

This study describes the architecture of neurons and individual dendritic arbors of thirteen intracellularly labeled thalamocortical projection neurons that respond to non-noxious stimuli from the primate (Macaca fascicularis or Macaca mulatta) ventrobasal complex (VB). The neurons compose a homogeneous morphological class with total dendritic lengths from 10,169 microns to 21,711 microns (mean 17,615 microns +/- 3,705). The labeled neurons were remarkably similar in most measured parameters including the number of dendrites (7.5 +/- 1.2), percentage of dichotomous branching (89.8% +/- 3.4), and contribution of terminal branches to total dendritic length (88.4% +/- 2.0). The individual dendrites ranged in total length from 443 microns to 7,657 microns with a mean of 2,346 microns (+/- 137, n = 98). There was a positive correlation between stem dendrite diameter and total dendrite length, making it possible to estimate the total size of an individual dendrite by measuring the stem dendrite diameter. There was only a small increase in mean path distance with increasing dendritic size at the whole neuron and individual dendritic levels, so that for individual dendrites the mean path distance of a dendrite consisting of only two segments was 199 microns, while the mean path distance for a dendrite with eight segments was only 45 microns longer. Analysis of dendrite diameter, segment order, and path distance shows that dendritic diameter is not reliable for determining the location of synaptic contacts viewed by electron microscopy onto dendritic trees. The small variation of measured parameters between these neurons presents a powerful tool for future developmental, plasticity and comparative studies of VB neurons.

Dendritic architecture of rat somatosensory thalamocortical projection neurons.

This study examines dendrites from physiologically characterized and intracellularly labelled thalamocortical projection (TCP) neurons from the rat ventrobasal complex (VB) and posterior nucleus (POm). The goals were to provide quantitative descriptions of TCP neuron dendrites, examine underlying design principles of dendritic morphology, and determine correlations between dendritic size parameters. Forty-four dendrites from seven VB neurons and 21 dendrites from three POm TCP neurons that responded to low-threshold mechanical stimuli were reconstructed and quantitatively analyzed at the light microscopic level. The dendritic architecture of the neurons was remarkably similar in most parameters studied, including the percentage of dichotomous branching, contribution of terminal branches to total dendritic length, and branching symmetry. There was a positive correlation between stem dendrite diameter and the length of the entire dendrite arbor, making it possible to estimate the total length of a dendritic arbor by measuring the stem dendrite diameter. The correlations of the VB and POm dendrites had different slopes. The path distance (the distance from the soma to a dendritic end point) of individual dendrites showed only a small variation with large differences in the total dendritic length of an arbor. The constant diameter of distal dendrites shows that dendrite diameter is a poor predictor of synaptic location on the dendritic tree. Although the morphology of neurons and their individual dendrites varied considerably in overall size and qualitative appearance, when examined qualitatively, many aspects of dendritic structure were similar within and between groups. We suggest that the rat somatosensory TCP neurons have a stereotyped dendritic architecture and present data which provide a base for future comparative, developmental, and plasticity studies.

Architecture of individual dendrites from intracellularly labeled thalamocortical projection neurons in the ventral posterolateral and ventral posteromedial nuclei of cat.

This study provides quantitative descriptions of individual dendrites from electrophysiologically characterized and intracellularly labeled thalamocortical projection (TCP) neurons of the cat ventrobasal complex. One hundred nine dendrites from six ventral posterolateral (VPL) neurons and six ventral posteromedial (VPM) neurons were examined. Measurement of several parameters showed that the individual dendrites were very similar to each other in overall architecture even though they varied greatly in total length and number of dendritic branches. The mean path distance (length from soma to a dendritic tip) was very similar for all dendrites in each group (VPL or VPM) regardless of the number of branches found along the path distance. However, VPL dendrites had a longer mean path distance (VPL = 206 +/- 36 microns; n = 51) than VPM dendrites (VPM = 182 +/- 29 microns; n = 58; P < 0.001). For all dendrites there was a strong correlation between the stem dendrite diameter and the dendritic length, which allows the estimation of dendritic length from dendrite diameter. Analysis of dendritic scaling shows that branches higher than first order do not follow Rall's 3/2 power rule, so these neurons cannot be modeled using the equivalent cylinder approximation. The data add to the qualitative descriptions of cat ventrobasal (VB) TCP dendrites currently available and provide a basis for future comparative, developmental, and plasticity studies. Analysis shows that many parameters of cat VB TCP dendrites fall within a narrow range, suggesting that, regardless of differences in length or superficial appearance, these dendrites share a stable underlying architecture.

Cell body and dendritic tree size of intracellularly labeled thalamocortical projection neurons in the ventrobasal complex of cat.

Fifteen thalamocortical projection (TCP) neurons from the adult cat ventrobasal complex (VB) were intracellularly labeled with horseradish peroxidase or neurobiotin and examined quantitatively. We find that cat TCP neurons share key morphological features and form one neuronal type. Previously reported variations in dendritic appearance cannot be supported by the present quantitative data. The number of dendrites varied between 4 and 13 (mean 9.1; +/- 4.0) and the total dendritic length of adult cat VB neurons varied between 9,421 and 19,646 microns (mean 13,120 microns; +/- 2,605). Linear regression analyses showed that soma diameter or cross-sectional area measurements provide a poor estimate for total dendritic length in TCP neurons. In contrast, the number of first order dendrites or the sum of first order dendrite diameters do provide a good estimate of overall TCP neuron size. This relationship is useful in predicting total dendritic length when it is not possible to reconstruct the entire dendritic tree. The mean dendritic path distance (distance from soma to the dendritic tip measured along the dendrite) was relatively constant for all neurons regardless of differences in total dendritic length or the number of branches that form the path distance.

Dendritic arbors of neurons from different regions of the rat thalamic reticular nucleus share a similar orientation.

Neurons in different regions of the rat thalamic reticular nucleus were labeled with biotin dextran amine and reconstructed. When viewed in coronal section, some neurons had a radial dendritic tree while others had dorso-ventrally elongated arbors. When rotated, all the neurons had a planar, disc-shaped dendritic field with the dendrites orientated parallel to the long axis of the nucleus. We conclude that all thalamic reticular nucleus neurons have a similar dendritic morphology and orientation.

Preserved features of thalamocortical projection neuron dendritic architecture in the somatosensory thalamus of the rat, cat and macaque.

A number of studies have shown that the organization of the mammalian somatosensory thalamus varies between species. As differences in cellular and synaptic thalamic organization would be expected to influence neuronal dendritic architecture, we compared somatosensory thalamocortical projection (TCP) neurons from the rat, cat and macaque. The results show that key features of the dendritic branching pattern remain unchanged despite large differences in the size of TCP neurons between the species. The features examined were: (i) ratio of the length of terminal branches to the length of the entire dendritic tree; (ii) the percentage of branch points that gave rise to two daughter branches as opposed to those that gave rise to three or more daughter branches; (iii) the proportional sum of absolute deviations (a measure of branching symmetry), and (iv) the mean branch order of the terminal segments. The present study provides evidence that somatosensory TCP neurons in these species comprise a homogeneous class and share a common dendritic architecture that is conserved across species despite changes in other aspects of thalamic circuitry. This suggests that TCP neuronal form is based on relatively stable genetic blueprint and that epigenetic factors (e.g. synaptic input) resulting from evolutionary changes in thalamic organization have had less influence on dendritic architecture.

Dendritic orientation of thalamocortical projection neurons in the ventrobasal complex of macaques.

This study describes the orientation of dendritic arbors from intracellularly labeled thalamocortical projection (TCP) neurons of Macaca fascicularis or Macaca mulatta. All neurons were located in the ventrobasal complex and responded to non-noxious stimuli. Each neuron was composed of dendrites that varied considerably in size and each dendrite tended to occupy a particular region of the perisomatic space with minimal overlap with other dendrites. Quantitative and qualitative analysis of the dendritic arbors of the labeled neurons showed they had an asymmetric distribution so that some regions of the perisomatic space contained more of the dendritic tree than others. Eleven of the thirteen reconstructed neurons had a larger percentage of the dendritic tree projecting into the medial portion of the perisomatic space. These results show that the dendritic arbors of macaque TCP neurons are not organized in a radially symmetric pattern as previously described and the asymmetric distribution of dendrites may be related to synaptic input.

Bilateral cavernous malformations resulting from cranial irradiation of a choroid plexus papilloma.

We report a case of multiple parietal cavernous malformations in a thirteen year old female who received cranial irradiation following incomplete resection of a choroid plexus papilloma. The cavernous malformations, which developed within the prior parietal radiation ports, were diagnosed nine years after the patients' radiation treatment when the patient presented with increasing frequency of seizures. Family history was negative for familial cavernous malformations. Due to the worsening frequency of seizures, the patient underwent resection of these two cavernous malformations with diagnosis confirmed by pathology. Post-operative, there was a significant reduction in seizure frequency. The origin and pathophysiology of cavernous malformations remains controversial. Cranial radiation treatment for tumors, particularly in children, may possibly lead to the development of these lesions, as occurred in this case. This is, to our knowledge, the first case of multiple cavernous malformations occurring within a previous radiation field following radiotherapy for a neoplasm.

Long-term effects of a lumbosacral ventral root avulsion injury on axotomized motor neurons and avulsed ventral roots in a non-human primate model of cauda equina injury.

Here, we have translated from the rat to the non-human primate a unilateral lumbosacral injury as a model for cauda equina injury. In this morphological study, we have investigated retrograde effects of a unilateral L6-S2 ventral root avulsion (VRA) injury as well as the long-term effects of Wallerian degeneration on avulsed ventral roots at 6-10 months post-operatively in four adult male rhesus monkeys. Immunohistochemistry for choline acetyl transferase and glial fibrillary acidic protein demonstrated a significant loss of the majority of the axotomized motoneurons in the affected L6-S2 segments and signs of an associated astrocytic glial response within the ventral horn of the L6 and S1 spinal cord segments. Quantitative analysis of the avulsed ventral roots showed that they exhibited normal size and were populated by a normal number of myelinated axons. However, the myelinated axons in the avulsed ventral roots were markedly smaller in caliber compared to the fibers of the intact contralateral ventral roots, which served as controls. Ultrastructural studies confirmed the presence of small myelinated axons and a population of unmyelinated axons within the avulsed roots. In addition, collagen fibers were readily identified within the endoneurium of the avulsed roots. In summary, a lumbosacral VRA injury resulted in retrograde motoneuron loss and astrocytic glial activation in the ventral horn. Surprisingly, the Wallerian degeneration of motor axons in the avulsed ventral roots was followed by a repopulation of the avulsed roots by small myelinated and unmyelinated fibers. We speculate that the small axons may represent sprouting or axonal regeneration by primary afferents or autonomic fibers.

The soma and proximal dendrites of sympathetic preganglionic neurons innervating the major pelvic ganglion in female rats receive predominantly inhibitory inputs.

Sympathetic preganglionic neurons (SPNs) in the intermediolateral (IML) and dorsal commissural nucleus (DCN) of the thoracolumbar segments of the spinal cord contribute to the autonomic control of the pelvic visceral organs. We examined the morphology of these neurons at the light and electron microscopic level and quantified the boutons apposing the soma and proximal dendrites of the SPNs innervating the major pelvic ganglion (MPG) in female rats. The majority of these cells resided in the DCN (61.6±6.2%) and IML (33.2±4.4%) nuclei. Measurements of cell volume and shape revealed no differences between SPNs sampled from the DCN and IML populations. Ultrastructural studies of DCN and IML SPNs revealed that coverage of SPNs by synaptic inputs is sparse, with an average of 11.60±2.41% of the soma membrane and 16.33±6.18% of proximal dendrites apposed by boutons, though some somata exhibited no synaptic coverage. Three distinct types of boutons were found to appose the SPN somata and dendrites. The putatively inhibitory F-type bouton covered a significantly greater percentage of membrane on the soma (8.48±2.12%) and dendrites (12.65±4.34%), than the S-type bouton, a putatively excitatory bouton, which only covered 2.94±0.70% of the somatic and 3.68±2.98% of the dendritic membranes. Boutons with dense-core vesicles were rare. Our results demonstrate that SPNs of the DCN and IML of female rats are similar morphologically, and that synaptic input on these cells, though sparse, is predominantly inhibitory.

Anatomical tracer injections into the lower urinary tract may compromise cystometry and external urethral sphincter electromyography in female rats.

Physiological and anatomical investigations are commonly combined in experimental models. When studying the lower urinary tract (LUT), it is often of interest to perform both urodynamic studies and retrogradely labeled neurons innervating the peripheral target organs. However, it is not known whether the use of anatomical tracers for the labeling of, e.g. spinal cord neurons may interfere with the interpretation of the physiological studies on micturition reflexes. We performed cystometry and external urethral sphincter (EUS) electromyography (EMG) under urethane anesthesia in adult female rats at 5-7 days after injection of a 5% fluorogold (FG) solution or vehicle into the major pelvic ganglia (MPG) or the EUS. FG and vehicle injections into the MPG and EUS resulted in decreased voiding efficiency. MPG injections increased the duration of both bladder contractions and the inter-contractile intervals. EUS injections decreased EUS EMG bursting activity during voiding as well as increased both the duration of bladder contractions and the maximum intravesical pressure. In addition, the bladder weight and size were increased after either MPG or EUS injections in both the FG and vehicle groups. We conclude that the injection of anatomical tracers into the MPG and EUS may compromise the interpretation of subsequent urodynamic studies and suggest investigators to consider experimental designs, which allow for physiological assessments to precede the administration of anatomical tracers into the LUT.

At-level neuropathic pain is induced by lumbosacral ventral root avulsion injury and ameliorated by root reimplantation into the spinal cord.

Neuropathic pain is common after traumatic injuries to the cauda equina/conus medullaris and brachial plexus. Clinically, this pain is difficult to treat and its mechanisms are not well understood. Lesions to the ventral roots are common in these injuries, but are rarely considered as potential contributors to pain. We examined whether a unilateral L6-S1 ventral root avulsion (VRA) injury in adult female rats might elicit pain within the dermatome projecting to the adjacent, uninjured L5 spinal segment. Additionally, a subset of subjects had the avulsed L6-S1 ventral roots reimplanted (VRA+Imp) into the lateral funiculus post-avulsion to determine whether this neural repair strategy elicits or ameliorates pain. Behavioral tests for mechanical allodynia and hyperalgesia were performed weekly over 7 weeks post-injury at the hindpaw plantar surface. Allodynia developed early and persisted post-VRA, whereas VRA+Imp rats exhibited allodynia only at 1 week post-operatively. Hyperalgesia was not observed at any time in any experimental group. Quantitative immunohistochemistry showed increased levels of inflammatory markers in laminae III-V and in the dorsal funiculus of the L5 spinal cord of VRA, but not VRA+Imp rats, specific to areas that receive projections from mechanoreceptive, but not nociceptive, primary afferents. These data suggest that sustained at-level neuropathic pain can develop following a pure motor lesion, whereas the pain may be ameliorated by acute root reimplantation. We believe that our findings are of translational research interest, as root implantation surgery is emerging as a potentially useful strategy for the repair of cauda equina/conus medullaris injuries.

Guidelines for the conduct of clinical trials for spinal cord injury as developed by the ICCP panel: spontaneous recovery after spinal cord injury and statistical power needed for therapeutic clinical trials.

The International Campaign for Cures of Spinal Cord Injury Paralysis (ICCP) supported an international panel tasked with reviewing the methodology for clinical trials in spinal cord injury (SCI), and making recommendations on the conduct of future trials. This is the first of four papers. Here, we examine the spontaneous rate of recovery after SCI and resulting consequences for achieving statistically significant results in clinical trials. We have reanalysed data from the Sygen trial to provide some of this information. Almost all people living with SCI show some recovery of motor function below the initial spinal injury level. While the spontaneous recovery of motor function in patients with motor-complete SCI is fairly limited and predictable, recovery in incomplete SCI patients (American spinal injury Association impairment scale (AIS) C and AIS D) is both more substantial and highly variable. With motor complete lesions (AIS A/AIS B) the majority of functional return is within the zone of partial preservation, and may be sufficient to reclassify the injury level to a lower spinal level. The vast majority of recovery occurs in the first 3 months, but a small amount can persist for up to 18 months or longer. Some sensory recovery occurs after SCI, on roughly the same time course as motor recovery. Based on previous data of the magnitude of spontaneous recovery after SCI, as measured by changes in ASIA motor scores, power calculations suggest that the number of subjects required to achieve a significant result from a trial declines considerably as the start of the study is delayed after SCI. Trials of treatments that are most efficacious when given soon after injury will therefore, require larger patient numbers than trials of treatments that are effective at later time points. As AIS B patients show greater spontaneous recovery than AIS A patients, the number of AIS A patients requiring to be enrolled into a trial is lower. This factor will have to be balanced against the possibility that some treatments will be more effective in incomplete patients. Trials involving motor incomplete SCI patients, or trials where an accurate assessment of AIS grade cannot be made before the start of the trial, will require large subject numbers and/or better objective assessment methods.

Transformation of synaptic vesicle phenotype in the intramedullary axonal arbors of cat spinal motoneurons following peripheral nerve injury.

Permanent transection of a peripheral motor nerve induces a gradual elimination of whole axon collateral systems in the axotomized spinal motoneurons. There is also an initial concurrent decrease in the amount of recurrent inhibition exerted by these arbors in the spinal cord for up to 6 weeks after the injury, whereas the same reflex action returns to normal by the 12-week postoperative state. The aim of the present investigation was to study the fine structure of the intramedullary axonal arbors of axotomized alpha-motoneurons in the adult cat spinal cord following a permanent peripheral motor nerve lesion. For this purpose, single axotomized alpha-motoneurons were labeled intracellularly with horseradish peroxidase at 12 weeks after permanent transection of their peripheral motor nerve. The intramedullary portions of their motor axon and axon collateral arbors were first reconstructed at the light microscopic level and subsequently studied ultrastructurally. This study shows that the synaptic contacts made by the intramedullary axon collateral arbors of axotomized motoneurons have undergone a change in synaptic vesicle ultrastructure from spherical and clear vesicles to spherical and dense-cored vesicles at 12 weeks after the transection of their peripheral axons. We suggest that the present transformation in synaptic vesicle fine structure may also correspond to a change in the contents of these boutons. This may, in turn, be responsible for the strengthening and recovery of the recurrent inhibitory reflex action exerted by the axotomized spinal motoneurons following a prolonged permanent motor nerve injury.

Neurofilamentous hypertrophy of intramedullary axonal arbors in intact spinal motoneurons undergoing peripheral sprouting.

An incomplete motor nerve injury or a partial loss of motoneurons leads to a partial denervation of skeletal muscle. As part of a compensatory response, the remaining intact motoneurons undergo peripheral sprouting and increase their motor unit size. Our knowledge about the responses in the more proximal parts of these sprouting motoneurons is sparse, however. We investigated the effects of an incomplete transection of the medial gastrocnemius (MG) nerve in the adult cat on the morphology of the intramedullary axon and axon collateral systems of the remaining intact MG motoneurons. At twelve weeks following the partial transection of the MG nerve, intracellular recording and labeling techniques were used to deposit horseradish peroxidase into single intact MG motoneurons for detailed morphological studies. The light microscopic appearance and caliber of the intramedullary stem motor axons of the intact MG motoneurons were indistinguishable from controls. The number and size of the intramedullary motoraxon collateral systems were also unchanged. However, frequent and marked hypertrophy of the distal portions of the motoraxon collaterals was encountered. Electron microscopic studies of the hypertrophied collaterals demonstrated abnormal accumulations of disorganized neurofilaments arranged in bundles or whorls. The morphological changes were indistinguishable from the neurofilamentous hypertrophy that has previously been reported in Wallerian degeneration, in experimental and human motor neuron disease and in some regenerating axonal processes of spinal motoneurons. We conclude that, neurofilamentous hypertrophy of the intramedullary arbors of motor axons may also be part of a reactive and non-degenerative response in intact motoneurons undergoing compensatory peripheral sprouting.