Spinal Gray Research Articles

A neural basis for motor primitives in the spinal cord.

Motor primitives and modularity may be important in biological movement control. However, their neural basis is not understood. To investigate this, we recorded 302 neurons, making multielectrode recordings in the spinal cord gray of spinalized frogs, at 400, 800, and 1200 mum depth, at the L2/L3 segment border. Simultaneous muscle activity recordings were used with independent components analysis to infer premotor drive patterns. Neurons were divided into groups based on motor pattern modulation and sensory responses, depth recorded, and behavior. The 187 motor pattern modulated neurons recorded comprised 14 cutaneous neurons and 28 proprioceptive neurons at 400 mum in the dorsal horn, 131 intermediate zone interneurons from approximately 800 microm depth without sensory responses, and 14 motoneuron-like neurons at approximately 1200 microm. We examined all such neurons during spinal behaviors. Mutual information measures showed that cutaneous neurons and intermediate zone neurons were related better to premotor drives than to individual muscle activity. In contrast, proprioceptive-related neurons and ventral horn neurons divided evenly. For 46 of the intermediate zone interneurons, we found significant postspike facilitation effects on muscle responses using spike-triggered averages representing short-latency postspike facilitations to multiple motor pools. Furthermore, these postspike facilitations matched significantly in both their patterns and strengths with the weighting parameters of individual primitives extracted statistically, although both were initially obtained without reference to one another. Our data show that sets of dedicated interneurons may organize individual spinal primitives. These may be a key to understanding motor development, motor learning, recovery after CNS injury, and evolution of motor behaviors.

Primary afferent terminals that express presynaptic NR1 in rats are mainly from myelinated, mechanosensitive fibers.

Presynaptic N-methyl-D-aspartate (NMDA) receptors in terminals of primary afferents to spinal cord of rats were first reported by Liu et al. (1994; Proc. Natl. Acad. Sci. USA 91:8383-8387) and were proposed to modulate nociceptive input (Liu et al. [1997] Nature 386:721-724). We previously demonstrated kainate and AMPA receptors in numerous primary afferent terminals in the spinal cord fixed with diluted paraformaldehyde and no glutaraldehyde. Therefore, we reinvestigated the occurrence of presynaptic NMDAR1 (NR1) with this fixation protocol. With confocal microscopy, numerous immunofluorescent puncta were double-stained for NR1 and the presynaptic marker synaptophysin throughout the spinal gray. NR1-immunostained puncta costained more frequently with a tracer that labels myelinated afferents (cholera toxin subunit B; CTB) than with a tracer that labels non-peptidergic unmyelinated afferents (Griffonia simplicifolia isolectin B4; IB4). Virtually no double staining was found for NR1 and calcitonin gene-related peptide (CGRP), which labels somatic peptidergic primary afferents. In the gracile nucleus, virtually all puncta labeled for CTB appeared immunopositive for NR1. At the electron microscopic level, most immunopositive terminals in spinal cord and gracile nucleus displayed morphological characteristics of endings of myelinated primary afferents. NR1 was presynaptic in 60-65% of all synapses in which it was expressed pre- or postsynaptically, or both, in spinal laminae I-IV. Estimates for the gracile nucleus were higher (80%). No presynaptic NR1 was found in the ventroposterior thalamus. Because of the relative sparsity of presynaptic NR1 in terminals in laminae I and IIo and in terminals of peptidergic unmyelinated afferents, it is suggested that presynaptic NMDA receptors play a more significant role in modulation of mechanosensitive, innocuous input than in nociception.

Exercise increases mRNA levels for adhesion molecules N-CAM and L1 correlating with BDNF response.

In situ hybridization was used to evaluate whether long-term moderate locomotor exercise, which up-regulates BDNF and TrkB levels in the spinal gray matter of the adult rat, similarly influences the expression of the cell adhesion molecules N-CAM and L1. Exercise doubled the level of N-CAM mRNA hybridization signal in the lumbar spinal gray. The increase in L1 mRNA was less consistent. N-CAM mRNA levels slightly increased in the white matter. BDNF mRNA levels also increased in cells of the ventral horn and the white matter due to the exercise. These results suggest that exercise-induced rearrangements of the spinal network involve N-CAM, L1 and BDNF, crucial in different aspects of synaptic plasticity and synapse formation.

Contributions of the medullary raphe and ventromedial reticular region to pain modulation and other homeostatic functions.

The raphe magnus is part of an interrelated region of medullary raphe and ventromedial reticular nuclei that project to all areas of the spinal gray. Activation of raphe and reticular neurons evokes modulatory effects in sensory, autonomic, and motor spinal processes. Two physiological types of nonserotonergic cells are observed in the medullary raphe and are thought to modulate spinal pain processing in opposing directions. Recent evidence suggests that these cells may modulate stimulus-evoked arousal or alerting rather than pain-evoked withdrawals. Nonserotonergic cells are also likely to modulate spinal autonomic and motor circuits involved in thermoregulation and sexual function. Medullary serotonergic cells have state-dependent discharge and are likely to contribute to the modulation of pain processing, thermoregulation, and sexual function in the spinal cord. The medullary raphe and ventromedial reticular region may set sensory, autonomic, and motor spinal circuits into configurations that are appropriate to the current behavioral state.

Reduction of GluR2 RNA editing, a molecular change that increases calcium influx through AMPA receptors, selective in the spinal ventral gray of patients with amyotrophic lateral sclerosis.

Enhancement of calcium influx through the alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA)/kainate receptor is a plausible mechanism underlying selective neuronal death in amyotrophic lateral sclerosis (ALS). The calcium conductance of the AMPA receptor is regulated by the GluR2 subunit that is edited at the glutamine/arginine residue site in the subunit assembly. We investigated the molecular changes of GluR2 mRNA in the spinal cord of ALS cases, those of cases with other neurological diseases, and those of normal cases using reverse transcription-polymerase chain reaction combined with restriction enzyme cleavage. We found that the editing efficiency was significantly lower only in the ventral gray of ALS cases (virtually 0% in 2 cases) than in any spinal region of the disease controls and normal controls. In addition, expression of GluR2 mRNA is lower in the ventral gray of the ALS cases and disease controls than in that of the normal controls. The above molecular changes of GluR2 mRNA in the ventral gray of ALS cases may enhance calcium influx through AMPA receptors, thereby promoting neuronal vulnerability. The decrement of GluR2 mRNA editing efficiency is unique to the ventral gray of ALS cases and may be closely linked to the etiology of ALS.

Urocortin Expression in Rat Brain: Evidence Against a Pervasive Relationship of Urocortin-Containing Projections With Targets Bearing Type 2 CRF Receptors

Histochemical and axonal transport methods were used to clarify the central organization of cells and fibers that express urocortin (UCN), a recently discovered corticotropin-releasing factor (CRF)-related neuropeptide, which has been proposed as an endogenous ligand for type 2 CRF receptors (CRF-R2). Neurons that display both UCN mRNA and peptide expression were found to be centered in the Edinger-Westphal (EW), lateral superior olivary (LSO), and supraoptic nuclei; lower levels of expression are seen in certain cranial nerve and spinal motoneurons and in small populations of neurons in the forebrain. Additional sites of UCN mRNA and peptide expression detected only in colchicine-treated rats are considered to be minor ones. UCN-immunoreactive projections in brain are predominantly descending and largely consistent with central projections attributed to the EW and LSO, targeting principally accessory optic, precerebellar, and auditory structures, as well as the spinal intermediate gray. Although neither the EW nor LSO are known to project to the forebrain, UCN-ir neurons in the EW were identified that project to the lateral septal nucleus, which houses a prominent UCN-ir terminal field. Although substantial UCN-ir projections were observed to several brainstem cell groups that express CRF-R2, including the dorsal raphe and interpeduncular nuclei and the nucleus of the solitary tract (NTS), most prominent seats of CRF-R2 expression were found to contain inputs immunopositive for piscine urotensin I, but not rat UCN. The results define a central UCN system whose organization suggests a principal involvement in motor control and sensorimotor integration; its participation in stress-related mechanisms would appear to derive principally by virtue of projections to the spinal intermediolateral column, the NTS, and the paraventricular nucleus. Several observations, including the lack of a pervasive relationship of UCN-ir projections with CRF-R2-expressing targets, support the existence of still additional CRF-related peptides in mammalian brain.

Inhibitory Effects of Ventral Signals on the Development of Brn-3.0-Expressing Neurons in the Dorsal Spinal Cord

Brn-3.0, a POU-domain transcription factor, is expressed in specific postmitotic neurons in the dorsal part of the neural tube which are among the first spinal cord neurons to appear in development. In the mature spinal cord, the Brn-3.0 cells form a numerous population of scattered neurons in the intermediate spinal gray. Ablation of the notochord in chick embryos extends the domain of Brn-3.0 expression into the ventral neural tube, while ectopic grafts of notochord tissue suppress Brn-3.0 expression. The notochord effects on Brn-3.0 expression are reproducedin vivoby the implantation of a local source of recombinant Shh protein. The down-regulation of Brn-3.0 expression in the dorsal spinal cord by the notochord and Shh contrasts with the known inductive effects of these ventral signals on the approximately simultaneous development of the spinal motor neurons. In cultured explants of neural plate from the region of the presumptive spinal cord, Brn-3.0 neurons develop in the absence of surface ectoderm and ventral midline tissue, suggesting that the Brn-3.0 phenotype may represent a “default” developmental pathway for early spinal cord neurons. Together these results advance the understanding of the mechanism of the generation of neuronal diversity in the developing vertebrate CNS.

Induction of c- fos gene expression by spinal cord transection in the rat

Sympathetic nerve activity is maintained after high spinal injury through circuits that remain in question. We evaluated patterns of c- fos gene induction as a monitor of spinal neurons responding to high spinal cord transection in the rat. Rats were anesthetized with isofluorane. Lower cervical or upper thoracic spinal segments were exposed, immersed in warm mineral oil and transected. Spinal cords were exposed but not transected in anesthetized controls. After 2.5 h, spinalized and control rats were perfused for immunocytochemistry. Cervical and thoracolumbar spinal segments and dorsal root ganglia were sectioned coronally. Tissues were incubated in primary, polyclonal antisera raised in rabbit or sheep against a peptide sequence unique to the N-terminal domain of Fos, and processed immunocytochemically. Neurons were induced to express Fos-like immunoreactivity (FLI), bilaterally, in the spinal gray, but not in primary sensory ganglia. Spinal cord transection induced neurons to express FLI in thoracic laminae I, IIo (outer substantia gelatinosa). Vre (lateral reticulated division), VII (lamina intermedia) and X, and the intermediolateral cell column. Lamina VIII was also labeled in spinal-injured but not in control animals. Immunolabeled nuclei were prominent in lumbar segments and were concentrated in the medial third of laminae I and IIo, and in laminae VII and X. Few cells were labeled in upper cervical or sacral segments. FLI was sparse in the spinal gray of controls and expressed mainly within the dorsal root entry zone of upper thoracic segments. Patterns of c- fos gene expression were site-specific and correlated with laminae that respond predominantly to noxious stimulation and that contain sympathetic interneurons. Laminae that are responsive to non-noxious stimuli and activated by walking, IIi, nucleus proprius, medial V and layer VI were not induced to express FLI. We conclude that neurons in specific spinal laminae that process high threshold afferents and that harbor neurons with sympathetic nerve-related activity are activated selectively by spinal cord transections. We hypothesize that peripheral afferents processed by spinal-sympathetic circuit neurons may regulate sympathetic discharge in the absence of supraspinal drive.

Axonal branching and termination of cervical reticulospinal neurons in the cat brachial segments

Axonal branching patterns in the brachial segments of cervical reticulospinal neurons (C-RSNs) were examined in cats using intraaxonal injection of horseradish peroxidase (HRP). Axons of these neurons were electrophysiologically identified by their projection to the lower cervical but not to the lumbar segments and monosynaptic activation after tectal and pyramidal stimulation. Six axons were stained up to terminals. Their stem axons descended in the ventral funiculus near the boundary of the spinal gray. The majority of collateral axons crossed lamina VIII and distributed terminals in the whole area of lamina VIII, the middle part of lamina VII, the lateral 2/3 of lamina VI, the ventral part of lamina V, and sparsely in the limb motor nuclei. Their possible functional role in head orienting movements is discussed.

Medullary visceral reflex circuits: local afferents to nucleus tractus solitarii synthesize catecholamines and project to thoracic spinal cord.

Visceral feedback circuits in lower brainstem were elucidated with retrograde tracers by mapping neurons that issue local projections to the general visceral afferent division of the nucleus tractus solitarii (NTS) and dorsomotor vagal nucleus (DMX) in adult male rats. In study 1, spinal and intramedullary afferents to the visceral-sensorimotor complex (NTS-X) were traced to contiguous populations of cell bodies arranged in cylindrical segmental organization. NTS-X afferents derive from curvilinear arrays of neurons that parallel the efferent radiations of the solitariotegmental tract. Newly discovered afferents arise from circumscribed cell groups in the dorsal reticular formation and periventricular zone. Another source was traced to a paraambigual cell column in the apex of the rostral ventrolateral reticular nucleus (n.RVL). In study 2, catecholaminergic afferents were initially defined with combined retrograde transport-immunocytochemical methods. Deposits of retrograde tracers into NTS-X transported to neurons containing tyrosine hydroxylase (TH) in the A1, C1, and C3 areas or phenylethanolamine N-methyltransferase (PNMT) in the C1 area of the n.RVL and C3 area. In study 3, it was revealed that NTS-X afferents arise, in part, as collaterals of thoracic reticulospinal neurons. Deposits of the retrograde fluorescent tracer Fluorogold into the upper thoracic cord and rhodamine-labeled microbeads into NTS-X transported to the same neurons within a subambigual locus in n.RVL and parts of nucleus raphe magnus. In study 4, dual retrograde tracer-immunocytochemical analysis demonstrated that catecholamines are synthesized by a subset of neurons in the n.RVL that issue collaterals to the NTS-X and thoracic cord. Double retrogradely labeled TH- or PNMT-immunoreactive cell bodies were restricted to the C1 area within a 450-microns column bordered rostrally by the facial nucleus and ventrally by the medullary subpial surface. We conclude that visceral reflex arcs are reciprocally organized. Targets of NTS projection are also sources of local NTS-X afferent innervation. Catecholaminergic and other local afferents from reticular formation, periventricular, and spinal gray may, via collaterals, simultaneously modulate visceral reflex excitability at the level of NTS and the outflow of autonomic and respiratory motoneurons.

Distribution and morphology of sacral spinal cord neurons innervating pelvic structures in Xenopus laevis.

Relatively little is known about the organization of neural input to pelvic viscera in amphibia. In this study, sacral spinal efferent neurons were labeled in Xenopus laevis frogs by application of horseradish peroxidase (HRP) to the tenth spinal nerve, to pelvic musculature, or to the pelvic nerve. DiI was applied to the pelvic nerve with similar results. Labeled spinal neurons were located in the intermediate gray or in the ventral horn. Neurons in the tenth dorsal root ganglion, but not in the spinal cord, were labeled after application of HRP or DiI to the pudendal nerve. The labeled neurons in the spinal cord intermediate gray were in a position comparable to that of the mammalian sacral parasympathetic nucleus (SPN). Two apparent subdivisions included 1) a medial cluster of cells with mediolaterally oriented dendrites and 2) a lateral group with dorsoventrally oriented dendrites. An intermediate group, not clearly classed with the other two, was also identifiable. In some cases, labeled tenth nerve primary afferents were seen in contact with efferent neurons of the intermediate gray. Labeled neurons in the ventral horn medial to the lateral motor column were small, with dendrites oriented mediolaterally, in a position comparable to that of the mammalian Onuf's nucleus. The peripheral targets of DiI-labeled pelvic nerve axons were the compressor cloaca muscle, cloaca, and bladder. DiI-labeled pudendal nerve axons distributed peripherally to cloacal lip and medial thigh integument. These data suggest that the pudendal nerve in amphibians is purely sensory and that both somatic and autonomic motor axons traverse the pelvic nerve.

Topographic specificity of corticospinal connections formed in explant coculture

The corticospinal pathway connects layer V pyramidal neurons in discrete regions of the sensorimotor cortex to topographically matching targets in the spinal cord. In rodents initial pathway errors occur transiently during early postnatal development, such that visual cortical axons project inappropriately into the corticospinal tract. Nevertheless, only sensorimotor axons form corticospinal connections, which are topographically ordered in hamsters from the earliest stages of innervation. Previous work in vivo suggests that pathfinding is carried out by primary cortical axons whereas target innervation occurs by extension of axon collaterals at appropriate locations. In vitro studies have provided evidence that chemotropic factors may selectively attract extension of neurites into specific targets. To investigate the basis for corticospinal target selection during development, we have used an in vitro explant coculture system. Sensorimotor and visual cortical explants from newborn hamsters were presented with inappropriate targets from olfactory bulb and cerebellum and targets from the cervical (forelimb) and lumbar (hindlimb) enlargements of the early postnatal spinal cord. Under in vitro conditions, corticospinal target selection was highly specific and remarkably similar to corticospinal connectivity in vivo. Visual and sensorimotor cortical neurites extended nonselectively into the white matter of the spinal cord. However, only neurites from the sensorimotor cortex were able to extend into and arborize within the spinal gray. In the majority of cases, these connections were topographically appropriate, matching forelimb cortex to cervical cord and hindlimb cortex to lumbar cord. However, we found no evidence that chemotropic attraction was responsible for selection of appropriate targets by cortical neurites or that spinal target tissue promoted extension of cortical axon collaterals within the collagen matrix. These results suggest that the ability of cortical neurites to recognize correct spinal targets and form terminal arbors may require direct axon target interaction.

Connections of the basal telencephalic areas c and d in the turtle brain.

Tracer substances were injected into the basal telencephalic areas c and d of the turtle brain. These areas (Acd) have recently been shown to be connected reciprocally with the dorsal spino-medullary region, though the particular subregions involved in these projections remained unclear. We demonstrated that the efferent projections of area d terminate predominantly within or immediately adjacent to the trigeminal nuclear complex and in the high cervical spinal gray. The dendritic domain of the vagus-solitarius complex and the dorsal column nuclear complex might also receive some basal telencephalic efferents. The afferent projections to Acd, on the other hand, arise predominantly in the dorsal column nuclei as defined according to cytoarchitectural and hodological criteria. A few retrogradely labeled cells were found in the vagus-solitarius complex, the principal trigeminal nucleus and the high cervical spinal cord. Numerous labeled cells were found in the dorsolateral isthmo-rhombencephalic tegmentum, especially the n. visceralis secundarius, the n. vestibularis superior and parts of the lateral lemniscal complex. Aminergic cell populations projecting to Acd were the n. raphes inferior and superior, the locus coeruleus, the substantia nigra, pars compacta and the ventral tegmental area. Other meso-diencephalic cell groups were the griseum centrale (including the n. laminaris of the torus semicircularis), the n. interpeduncularis dorsalis, the nucleus of the fasciculus longitudinalis medialis, the nucleus and the nucleus interstitialis of flm, the n. interstitialis commissuralis posterior and then n. caudalis. Several hypothalamic regions, the reuniens complex and the perirotundal region of the thalamus also appeared to project heavily to Acd. Telencephalic areas retrogradely labeled after injection of tracer into Acd and its immediate surroundings were the rostral part of the lateral (olfactory) cortex, adjacent regions of the basal dorsal ventricular ridge and the n. centralis amygdalae, the n. tractus olfactorius lateralis as well as the areas g and h. The data suggest that areas c and d may correlate best with the 'extended' amygdala in mammals; further correlation with structures similar to the ventral striopallidum, however, cannot be excluded. Homostrategies are discussed with regard to the processing of higher-order somatovisceral information in turtles, birds and mammals.

Tropism and corticospinal target selection in the rat

Layer V pyramidal cells in the intermediate part of the cerebral cortex enter the lumbar spinal gray, but not the cervical spinal gray matter, during the first postnatal week. To study if the ingrowth of intermediate corticospinal axons into the lumbar spinal gray is guided by a diffusible tropic factor, we co-cultured expiants of the intermediate part of the sensorimotor cortex and of the lumbar spinal gray matter in 3-D collagen gels. Using this test system, a target specific directional growth of cortical axons towards the lumbar spinal gray explant can be demonstrated in vitro. Retrograde labeling indicates that most labeled cell bodies were located in layer V of the cortex explant and were characterized by a pyramidal cell shape. Furthermore, axon behavior of retrogradely labeled neurons within the cortical explant is considerably affected by the presence of lumbar spinal gray target tissue. In contrast to lumbar spinal gray innervation, intermediate corticospinal tract axons do not enter the cervical spinal gray in vivo. Is it the inability of intermediate corticospinal tract axons to respond to cervical target-derived influences? In the current study we co-cultured explants of the intermediate cortex and cervical spinal gray matter in 3-D collagen gels. Our data indicate that in vitro axons from layer V neurons in the intermediate part of the cortex are capable of recognizing and responding to a diffusible factor released by the cervical spinal cord target area. This suggests that the failure of neurons within the intermediate cortex to enter the cervical spinal gray in vivo, is not due to their inability to respond to a target-derived diffusible factor, but probably regulated by other extrinsic factors.

Neurogenesis of spinothalamic and spinocerebellar tract neurons in the lumbar spinal cord of the rat

The temporal and spatial neurogenic patterns of spinothalamic and spinocerebellar neurons were determined in spinal cord segment L1 of the rat. Neurogenic patterns were demonstrated with [ 3H]thymidine administered to fetal rats during the period when neurons with supraspinal projections are known to be generated, i.e. on one of embryonic (E) days E13, E14, or E15. The animals were allowed to survive 50 to 100 days postpartum, then neurons with spinothalamic and spinocerebellar projections were retrogradely filled with fluorescent axonal tracers, Fluoro-Gold or True blue, which were pressure injected into the dorsal thalamus and cerebellum in various combinations in the same and in separate animals. Neurons labeled with each retrograde tracer and [ 3H]thymidine and neurons labeled with retrograde tracers alone were counted in spinal cord segment L1 in each of the animals. Spinothalamic and spinocerebellar neurons were found to be separate and distinct populations. Statistical analysis of the data showed that spinothalamic and spinocerebellar neurons also have distinctly different patterns of neurogenesis which suggest early determination in each cell line. The temporal neurogenic pattern followed a projection-distance gradient, such that spinothalamic neurons, which have longer axons than spinocerebellar neurons, completed neurogenesis prior to spinocerebellar neurons in each region of the spinal gray.

Projections from the nucleus tractus solitarii to the spinal cord.

Projections from the nucleus tractus solitarii (NTS) to the spinal cord were demonstrated in the male Sprague-Dawley rat. In retrograde transport studies, a horseradish peroxidase conjugate or a fluorescent dye, FluoroGold, were injected into midcervical or upper thoracic spinal segments. Most solitariospinal neurons were multipolar or bipolar and located between the obex and spinomedullary junction. Solitariospinal neurons were concentrated in proximity to the ventral border of the solitary tract and extended dorsally into the intermediate division and ventrolaterally into the intermediate reticular zone (IRt) of the lateral tegmental field. This subgroup predominantly projects to midcervical spinal segments. A subset of small neurons was retrogradely labeled from cervical or thoracic spinal segments in the medial commissural nucleus and contiguous with a periventricular group surrounding the central canal. In anterograde transport studies, iontophoretic deposits of Phaseolus vulgaris leucoagglutinin were centered stereotaxically on sites in NTS identified by retrograde transport data. The lectin was incorporated by neurons of the solitary complex and transported bilaterally by axons that emerged from the nucleus and entered the reticular formation. The solitario-reticular (transtegmental) pathway irradiated diagonally across the IRt and extended caudally into the cervical lateral funiculus and spinal gray. A small periventricular-spinal pathway also descended longitudinally to the neuraxis. Solitariospinal neurons project to superficial lamina of the dorsal horn, laminae VII and X and ventral horn. The projections are predominantly contralateral to phrenic and intercostal motor nuclei and ipsilateral to the intermediolateral cell column. The solitariospinal projection represents the shortest route in the central nervous system, other than the local intraspinal reflex, through which first order visceral afferents signal cardiorespiratory and alimentary motor nuclei.

Neuronal types in the spinal dorsal gray of the turtle Chrysemys d'orbigny: a Golgi study

At thoracic and lumbar levels the spinal dorsal gray of young specimens of the turtle Chrysemys d'orbigny consists of a cell-free neuropil and an aggregation of perikarya termed here the lateral column of the dorsal horn (LCDH). Nerve cell clusters also occur in the dorsal commissure. The main neuropil area can be divided into a thin superficial layer containing some myelinated fibers (neuropil area Ib) and a compact core composed of unmyelinated axon terminals, dendritic branches, and thin glial processes (neuropil area II). A looser neuropil area is located at the horn base (neuropil area III). The so-called marginal zone of de Lange represents a fourth synaptic field termed here neuropil area Ia. The LCDH consists of neurons of different size and shape. Two peculiar nerve cell types have been recognized in the dorsal horn: giant and bitufted neurons. The former exhibits a large dendritic arbor, which after passing through neuropil areas II and Ib projects into neuropil area Ia and the adjacent white matter. Most frequently Golgi-stained giant neurons have perikarya and dendritic domains on the same side (ipsilateral giant neurons). There are also heterolateral giant neurons whose dendritic branches invade the opposite horn. Bitufted neurons are characterized by the presence of two main dendritic shafts connecting neuropil area II of both dorsal horns. At neuropil levels the major dendritic branches ramify profusely giving rise to short tortuous terminal processes. Perikarya of bitufted neurons occur in the dorsal commissure. The LCDH also contains many small and medium-sized neurons. These are oriented in two main directions: parallel or radial with respect to the dorsal horn surface. The population of horizontally oriented neurons comprises two subtypes termed here alpha and beta. Radially oriented neurons are pleomorphic, defying precise, unequivocal classification.

Postnatal development of the ipsilateral corticospinal component in rat spinal cord: A light and electron microscopic anterograde HRP study

Most developmental and regeneration studies using the corticospinal tract (CST) as a model are focussed on its major contralateral component in the ventralmost part of the dorsal funiculus. However, for a correct interpretation of the data in these and future studies, a complete anatomical description of the rat CST, including its ipsilateral uncrossed component, is a prerequisite. In this study we used anterograde horseradish-peroxidase (HRP) tract-tracing at the light and electron microscopic level to describe the development of the uncrossed ipsilateral CST component. In the far caudal portions of the medulla oblongata, most of the CST fibers decussate dorsally and medially to assume positions in the ventralmost part of the dorsal funiculus. However, a small number of labeled fibers continue uncrossed in a position just lateral to the ventral median fissure. Labeled ipsilateral CST fibers caudally extend into the ventral funiculus of the third thoracic segment at postnatal day 1 (P1), into the seventh thoracic segment at P3, and into the first lumbar segment at P5 and P7. After the first postnatal week labeled ipsilateral CST fibers gradually disappear from upper lumbar, lower-, and mid-thoracic spinal cord levels, respectively. From P14 through adulthood the ipsilateral CST projection can not be visualized below upper thoracic spinal cord levels. Quantitative analysis of the amount of label along the length of the outgrowing ipsilateral CST revealed a characteristic staggered pattern of outgrowth: a small number of fibers take the lead and additional fibers are successively added. The outgrowth of labeled ipsilateral CST fibers is mainly restricted to spinal cord white matter. Occasionally a few outgrowing fibers into the adjacent cervical spinal gray can be observed, however, only between P3 and P7. Electron microscopic observations at the cervical enlargement (fifth cervical segment) demonstrate that the first labeled ipsilateral CST fibers, at P1, are characterized by the presence of growth cone-like enlargements at their distal endings. The process of myelination of labeled ipsilateral CST axons starts at about day P14. Although myelination seems to be largely completed at P28, labeled unmyelinated axons are still present in the adult ipsilateral CST component.

Biochemical characterization and regional quantification of μ, δ and κ opioid binding sites in rat spinal cord

The spinal cord contains μ, δ and κ opioid receptors which mediate the antinociceptive effects of opioid agonists administered onto the spinal cord. In this study, we characterized the binding sites for highly-selective μ, δ and κ opioid radioligands and quantified and distribution of opioid binding sites in rat lumbosacral spinal cord using autoradiography. In sections of rat brain mounted on glass slides, the μ ligand, [ 3H]sufentanil, bound with high affinity with an apparentK d of 0.46 nM. The δ ligand, [ 3H]DPDPE ([ d-Pen 2,5]-enkephalin), bound with aK d of 4.31 nM, and the κ-ligand, [ 3H]U69593, bound with aK d of 2.27 nM. Three regions of the spinal gray were targeted for quantification of binding sites by autoradiography. The data indicate that when considered as a percentage of the total opioid binding capacity within a region, the contribution of μ sites in laminae I-II was about 90%, with δ and κ sites 7% and 3%, respectively. In lamina V, the μ sites comprised about 70% of the total opioid sites, with δ and κ sites comprising 28% and 2%, respectively. In the area adjacent to the central canal, μ sites contributed about 65% of the total opioid sites followed by δ sites at 33% and κ sites at 2% of total opioid sites. These results demonstrate a differential distribution of μ, δ and κ binding sites with respect to the organization of the spinal gray matter. The preferential occurrence of all 3 opioid binding sites in the superficial dorsal horn is noteworthy since many fine caliber primary afferent fibers mediating nociception establish synaptic contact in this region.

The recovery of forelimb-placing behavior in rats with neonatal unilateral cortical damage involves the remaining hemisphere

Following unilateral lesions of the somatic sensorimotor cortex (SMC) in neonatal, but not adult, rats, an aberrant ipsilateral corticospinal projection originates from the undamaged hemisphere (Hicks and D'Amato, 1970; Leong and Lund, 1973; Castro, 1975). We have evaluated the contribution of the hemisphere contralateral to a unilateral lesion of the SMC in the recovery of tactile forelimb-placing behavior. Neither adult-lesioned or neonatally lesioned animals show evidence for placing deficits with either forelimb when tested 30 or 42 d after the lesion. However, in adult-lesioned animals, a subsequent lesion of the undamaged SMC on postlesion day 42 produces placing deficits only with the forelimb contralateral to the second lesion, while such a second lesion in the neonatally lesioned rats results in placing deficits with both forelimbs. Anatomical observations in the animals used for behavioral analyses confirm previous reports of a substantial ipsilateral corticospinal projection in rats with unilateral SMC damage as neonates and demonstrate that many of these aberrant fibers recross the midline within the spinal cord to arborize extensively within the ipsilateral spinal gray. These findings indicate that, following unilateral SMC lesions in neonates, the contralateral hemisphere mediates some aspects of the recovery of forelimb placing. The aberrant ipsilateral corticospinal projection may provide the anatomical substrate through which the cortex effects this recovery.