Projections from the cervical enlargement to the cerebellar nuclei in the rat,studied by anterograde axonal tracing

Spinocerebellar projections from the cervical enlargement originate from neurons in the medial part of lamina VI and the central part of lamina VII. In the present study, the topographic projections of the cervical enlargement to the cerebellar nuclei were examined by anterograde tracing with biotinylated dextran in the rat. Following injections of the tracer into the spinal cord at levels between the C5 and T1 segments, anterogradely labeled axons and terminals were immunohistochemically demonstrated in the cerebellar nuclei. Unilateral injections revealed that projections are bilateral, but predominantly ipsilateral, to the cells of origin. Labeled axons entered the medial nucleus from its rostrodorsal and rostromedial aspects. Labeled terminals were distributed to dorsal and medial parts of the middle subdivision at its rostral levels and to medial parts of the caudomedial subdivision of the medial nucleus. Most axons terminated in the middle subdivision. Single axons were seen to course rostrocaudally in the medial nucleus and give off terminal axons to both subdivisions. A few labeled terminals were seen in the dorsolateral protuberance of the medial nucleus, and in the anterior interpositus and the posterior interpositus nuclei. No labeled terminals were seen in the lateral cerebellar nucleus. The present study demonstrates that spinocerebellar neurons in laminae VI and VII of the cervical enlargement project to dorsomedial areas of the medial nucleus at rostral levels, bilaterally but predominantly ipsilaterally. It is suggested that these areas specifically receive cutaneous and muscular input related to the forelimb movement. J. Comp. Neurol. 377:251–261, 1997. © 1997 Wiley-Liss, Inc.     

Distribution of spinal cord projections from the medullary subnucleus reticularis dorsalis and the adjacent cuneate nucleus: A phaseolus vulgaris- leucoagglutinin study in the rat

The distribution and organization of descending spinal projections from the dorsal part of the caudal medulla were studied in the rat following injections of Phaseolus vulgaris leucoagglutinin into small areas of the subnucleus reticularis dorsalis (SRD) and the adjacent cuneate nucleus (Cu). The caudal aspect of the Cu projected only to the dorsal horn of the ipsilateral cervical cord via the dorsal funiculus. These projections were mainly to laminae I, IV, and V. More ventrally located reticular structures projected to the full length of the cord. Fibers originating from the SRD travelled through the ipsilateral dorsolateral funiculus and terminated within the deep dorsal horn and upper layers of the ventral horn, mainly in laminae V–VII. Fibers originating from subnucleus reticularis ventralis (SRV) travelled ipsilaterally through the lateral and ventrolateral funiculi and bilaterally through the ventromedial funiculus. These fibers terminated within the ventral horn. The density of labeling within the gray matter varied at different levels of the cord was as follows: cervical > sacral > thoracic > lumbar. The reciprocal connections between the caudal medulla and the spinal cord suggest that the former is an important link in feedback loops that regulate spinal outflow. © 1995 Wiley-Liss, Inc.     

Organization of diencephalic projections from the medullary subnucleus reticularis dorsalis and the adjacent cuneate nucleus: A retrograde and anterograde tracer study in the rat

The distribution and organization of diencephalic projections from the subnucleus reticularis dorsalis (SRD) and the neighbouring cuneate nucleus (Cu) were studied in the rat by using microinjections of Phaseolus vulgaris leucoagglutinin in SRD and Cu and wheat germ agglutinin-apo horseradish peroxidase-gold in some selected thalamic areas. As previously reported, the efferent projections from the Cu were essentially contralateral and terminated mainly in the ventroposterolateral thalamic nucleus. Less dense terminals from the Cu were also observed in the posterior thalamic group, the ventral aspect of the zona incerta and the caudal and dorsal portion of the reuniens area. Retrograde tracer injections in the medial ventroposterolateral thalamic nucleus labeled numerous cells in the contralateral Cu, with a smaller number in the gracile nucleus. From the SRD, terminals were observed in the lateral aspect of the ventromedial thalamic nucleus, the lateral parafascicular area and, to a lesser extent, in the ventral aspect of the zona incerta and the core of the reuniens area. Retrograde tracer injections in the lateral part of the ventromedial thalamic nucleus labeled cells in the caudal medulla, many of which were located in the dorsal-most aspect of the SRD throughout its caudo-rostral extent. The existence of SRD-thalamic connections reinforces the idea that the caudal reticular formation is an important nociceptive relay to the thalamus. Our data shed new light on old hypotheses suggesting that, in addition to spino-thalamic pathways, spino-reticulo-thalamic pathways may play an important role in distributing pain signals to the forebrain. J. Comp. Neurol. 390:133–160, 1998. © 1998 Wiley-Liss, Inc.     

The projection of the medial and posterior articular nerves of the cat's knee to the spinal cord

We studied the spinal projections of the medial and posterior articular nerves (MAN and PAN) of the knee joint in the cat with the aid of the transganglionic transport of horseradish peroxidase. The afferent fibers of the MAN entered the spinal cord via the lumbar dorsal roots L5 and L6 and those of the PAN entered via the dorsal roots L6 and L7. Within the dorsal root ganglia, most labeled neurons had small to medium diameters. A relatively higher number of medium-size cell bodies were labeled from the PAN than from the MAN. In the spinal cord labeled MAN afferent fibers and terminations were most dense in the L5 and L6 segments, and those of the PAN were most dense in L6 and L7, that is, in the respective segments of entry. Labeled afferent fibers from both nerves projected rostrally at least as far as L1 and caudally as far as S2. Labeled fibers were found in Lissauer's tract as well as in the dorsal column immediately adjacent to the dorsal horn. In the spinal gray matter, both nerves had two main projection fields, one in the cap of the dorsal horn in lamina I, the other in the deep dorsal horn in laminae V-VI and the dorsal part of lamina VII. Both nerves, but particularly the PAN, projected to the medial portion of Clarke's column. No projection was found to laminae II, III, and IV of the dorsal horn or to the ventral horn. Since these findings parallel observations on hindlimb muscle afferent fibers, the present data support the existence of a common pattern for the central distribution of deep somatic afferent fibers.     

Anatomical organization of the spinocerebellar system in the cat,as studied by retrograde transport of horseradish peroxidase

The distribution of spinocerebellar tract (SCT) neurons has been studied in the entire length of the spinal cord of the cat following injections of horseradish peroxidase into the cerebellum, and whether or not the axons of the labeled neurons crossed within the spinal cord was determined in cases with injections preceded by hemisections at the cervical levels. The SCTs were classified into the following corssed and uncrossed tracts according to the cell origin and the fiber course; The crossed SCTs originate from (1) the central cervical nucleus (the CCN-SCT), (2) lamina VIII neurons of the cervical to the lumbar cord (the lamina VIII-SCT), (3) spinal border cells (the border cell-SCT), (4) neurons in the medial lamina VII of the lumbar to the caudal spinal segments (the medial lamina VII-SCT), (5) ventral horn neurons (laminae VII and VIII) of the sacral and caudal segments (the ventral horn-SCT) and (6) dorsal horn neurons (lamina V) of the sacral and the caudal segments (the dorsal horn-SCT). The uncorssed tracts originate from (1) neurons of the medial lamina VI of C2 to T1 (the medial lamina VI-SCT of the cervical cord), (2) neurons in the central part of lamina VII of C6 to T1 (the central lamina VII-SCT of the cervical enlargement), (3) lamina V neurons of the lower cervical to the lumbar cord (the lamina V-SCT), (4) Clarke's column (the Clarke's column-SCT) and (5) neurons in the medial lamina VI of L5 and L6 (the medial lamina VI-SCT of the lumbar cord). The present study suggests that the spinocerebellar system originates from more diverse laminae than has previously been known, and further refined studies on the topographic projections of each tract will yield more important and valuable information in this field.     

Vasoactive intestinal polypeptide and substance P in primary afferent pathways to the sacral spinal cord of the cat

An analysis of vasoactive intestinal polypeptide immunoreactivity (VIP-IR) and substance P-IR in the cat spinal cord has revealed marked differences in the distribution of the two peptides. While substance P-IR was located at all levels of the cord, VIP-IR was most prominent in the sacral segments in Lissauer's tract and lamina I on the lateral edge of the dorsal horn. VIP-IR was also present in the sacral cord in (1) laminae V, VII, and X, (2) a thin band on the medial side of the dorsal horn, (3) the dorsal commissure, (4) the lateral band of the sacral parasympathetic nucleus, and (5) in a few animals in Onuf's nucleus. In other segments of the spinal cord VIP-IR was much less prominent but was present in Lissauer's tract and laminae I, II, and X. Substance P-IR was more uniformly distributed at all segmental levels in laminae I-III, V, VII, and X and in the dorsal commissure. In ventrolateral lamina I of the sacral spinal cord both VIP-IR and substance P-IR exhibited a distinctive periodic pattern in the rostrocaudal axis. The peptides were associated with bundles of dorsoventrally oriented axons and varicosities spaced at approximately 210-micron intervals center to center along the length of the spinal cord. The bundles in lamina I continued into lamina V where they further divided into smaller bundles that extended medially through laminae V and VII. The most prominent bundles of VIP axons passed ventrally from lateral laminae V and VII to enter lamina X and the ventral part of the dorsal gray commissure. On the other hand the majority of substance P axons in lamina V turned dorsally to join with axons on the medial side of the dorsal horn and to pass into the dorsal part of the dorsal gray commissure. Rostrocaudal VIP axons were present not only in Lissauer's tract but also in dorsolateral lamina I, in the lateral funiculus and in the ependymal cell layer of the central canal. Following unilateral transection of the sacral dorsal roots (2 weeks-22 months) the density of VIP axons and terminals was markedly reduced in ipsilateral Lissauer's tract and lateral laminae I and V; however, no change was detected in lamina X. Sacral deafferentation reduced substance P-IR in the dorsal gray commissure and in lateral laminae I and V.(ABSTRACT TRUNCATED AT 400 WORDS)     

Uncrossed and crossed projections from the upper cervical spinal cord to the cerebellar nuclei in the rat, studied by anterograde axonal tracing

In the upper cervical spinal segments, neurons in the medial part of lamina VI give rise to uncrossed spinocerebellar axons, whereas the central cervical nucleus (CCN) and neurons in laminae VII and VIII give rise to crossed spinocerebellar axons. Using anterograde labeling with biotinylated dextran in the rat, we examined the projections of these neuronal groups to the cerebellar nuclei. Uncrossed and crossed projections were distinguished by cerebellar lesions placed on the side contralateral or ipsilateral to the tracer injections confined to the second and third cervical spinal segments (C2 and C3, respectively). Labeled terminals of uncrossed projections were seen in the middle, dorsal, and ventrolateral parts of the middle subdivision and in the ventral part of the caudomedial subdivision of the medial nucleus. In the anterior interpositus nucleus, terminals were seen in the middle of the mediolateral extent, whereas, in the posterior interpositus nucleus, they were seen in lateral and caudal parts. The terminals of crossed projections from the CCN were distributed ventrally in medial to ventrolateral parts of the middle subdivision of the medial nucleus. Some terminals were seen in the caudomedial subdivision of the medial nucleus. In the anterior interpositus nucleus, labeled terminals were seen mainly in rostromedial parts, whereas, in the posterior interpositus nucleus, they were seen in caudal and dorsal parts of the medial half. The present study suggests that the medial lamina VI group and the CCN in the upper cervical segments project to the different areas of the cerebellar nuclei and are concerned with different functions.     

Cells of origin of long descending propriospinal fibers connecting the spinal enlargements in cat and monkey determined by horseradish peroxidase and electrophysiological techniques.

The cells of origin of the long descending propriospinal tract (LDPT) in the cervical enlargement were studied in cat and monkey by using the retrograde transport of horseradish peroxidase (HRP). Their distribution was confirmed electrophysiologically in cat by recording their antidromic action potentials. In cats and monkeys unilateral injections of HRP were made into the gray matter of the lumbosacral enlargement, but there was some spread to the contralateral side. In cats labeled somas were found in greatest numbers in lamina VIII and medial lamina VII, bilaterally. Labeled cells also were found bilaterally in laminae I, IV--VI, and X, but few were in IV and VI. Those in lamina V were usually in the lateral part of the lamina near the reticulated region. The cross-sectional areas of 20 neurons from each of laminae I and V--VIII were measured. Cells in lamina I were smallest and the largest were in VII and VIII. In cats with the spinal cord hemisected between the injection site and the cervical enlargement containing the somas, the bilaterality of the LDPT neurons in laminae VII and VIII was confirmed anatomically and physiologically. Contralaterally projecting neurons in laminae VIII and medial VII constituted a majority of LDPT cells in those laminae. The LDPT neurons in the dorsal horn appeared to project mainly ipsilaterally, but the number of labeled dorsal horn cells in these preparations was small. The distribution of antidromically localized cells of the LDPT was found to be in good agreement with the anatomical results. Their conduction velocity was 59 +/- 22 m/s (mean +/- s.d., n = 245). Histograms of the conduction velocity by laminae are given. In monkey the distribution of labeled somas was similar to that in the cat, except that the concentration of labeled somas in the ventral horn was more medially and dorsally located. Labeled somas were found bilaterally in laminae I, IV--VIII, and X, but more appeared to be ipsilateral to the side of the injection, especially in the dorsal horn. The bilaterality of the LDPT in the monkey was not tested with hemisections of the spinal cord. Neurons of the LDPT are ideally situated for conveying sensory information from the forelimb for eliciting reflexes in the hindlimb, as has been observed after stimulating afferents in the forelimb, and for coordinating, in general, motor functions between the two pairs of limbs.     

Anatomical organization of long ascending propriospinal neurons in the cat spinal cord

Retrograde transport of lectin-HRP conjugate (WGA-HRP) was used to examine the anatomical organization of long ascending propriospinal neurons (LAPNs) projecting to the cervical enlargement (C5-T1) and to the upper part of the cervical cord (C3-4) in cats. Small injections (0.05-1.0 microliter) of dilute (1-4%) WGA-HRP were made into the C5-T1 or C3-4 regions. The field potential evoked from stimulation of the superficial radial nerve served to position the micropipette delivering injections. Small and localized populations of labelled LAPNs were found in the dorsal horn (laminae IV-V), the intermediate zone (dorsal and medial lamina VII), and the ventral horn (ventral lamina VII, laminae VIII and IX). Ventral horn LAPNs projecting to the C5-T1 region were preferentially located in rostral lumbar regions. Ventral LAPNs projecting to the C3-4 region were more caudally situated. No regional differences in distribution of dorsal horn and intermediate zone LAPNs were noted in comparing the results of C3-4 with C5-T1 injection protocols. It is concluded that the caudally located ventral LAPNs may exert their influence on cervical motor output through C3-4 propriospinal interneurons. Other LAPNs are considered to exert their effect more directly, either at the C5-T1 or the C3-4 levels.     

The terminations of corticospinal tract axons in the macaque monkey

This study examined the corticospinal tract in monkey by utilizing the anterograde transport of wheat germ lectin conjugated to horseradish peroxidase (WGA HRP) at the light microscopic level and the axonal transport of 3H-proteins with both light and electron microscopic autoradiographic techniques. The animals survived 3-9 days after the injections of 3H-leucine or 3H-leucine/WGA HRP into either motor or sensory cortices. With the laminar schema of Rexed as a guide to the layers of the spinal gray matter, qualitative and quantitative analyses of labeled projections of the corticospinal tract (CST) were made. With the light microscope, axons from the sensory cortex labeled with WGA-HRP could be observed in the contralateral spinal gray from lamina I to the border of laminae VI/VII, the heaviest distribution being located in medial III-VI. There was a small ipsilateral projection to V and VI. With 3H label, laminae I and II revealed few overlying silver grains; many grains overlay laminae III-VI. Projections from the motor cortex labeled with either WGA-HRP or 3H extended from the contralateral laminae III/IV border into the motor nucleus (lamina IX) and were seen to be somewhat more dense in the lateral areas of the spinal gray. The motor cortex projected heavily to ipsilateral VIII, and in sparse amounts to ipsilateral V and VI. Electron microscopy of radioactive axons from the sensory cortex to dorsal horn revealed many radioactive myelinated fibers and some labeled non-myelinated axons. Labeled terminals contacted medium to small dendrites; there were a few labeled C-type profiles in glomeruli and occasional axo-axonal or dendro-axonal contacts, the labeled cortical axons being the postsynaptic structure. In ventral horn following motor cortex injections, the labeled axons were all myelinated. The synaptic contacts were found on small, medium, and large proximal dendrites as well as on cell bodies. Labeled terminals which formed the central element in glomeruli were also seen in this region. Most of the labeled corticospinal terminals in dorsal and ventral horn contained rounded vesicles, but a significant number revealed pleomorphic vesicles. The relationship of these morphological findings to physiological studies of the CST is presented.     

The distribution of dorsal root axons to laminae IV,V, and VI of the macaque spinal cord: A quantitative electron microscopic study

The projections of dorsal root axons to the deeper laminae (IV, V, and VI) of the Macaque spinal cord were examined by the use of experimentally induced degeneration following dorsal rhizotomy or by injection of dorsal root ganglia with tritiated amino acids followed by light and electron mi-croscopic autoradiography. Following dorsal rhizotomy, neurofilamentous degeneration of synaptic profiles occurs in each of the three deep laminae, more commonly in laminae IV and V than in lamina VI. The neurofilamentous degeneration is seen both in central glomerular (C) profiles and in many of the round vesicle (R) profiles. Neurofilamentous degeneration occurs as early as 18 hours following rhizotomy and the degenerating terminals are most numerous at 3–4 days postrhizotomy. None are seen after 7 days survival. The neurofilamentous profiles form axodendritic and, occasionally, axosomatic synapses with neurons of the dorsal horn. They are also seen to be postsynaptic to flat vesicle (F) profiles in axoaxonal synapses. A second type of degeneration, electron-lucent degeneration, is seen in laminae V and VI, and only occasionally in lamina IV. The lucent degeneration occurs somewhat later after rhizotomy than does the neurofilamentous degeneration and reaches its peak at 5 days postrhizotomy. No lucent terminals are seen after 7 days survival. Electron-dense degeneration, so common in lamina II, is not seen in the deeper dorsal horn. Autoradiographic techniques show that both C and R terminals are labelled in the deeper dorsal horn. Both of these terminals form axodendritic synapses and a significant number are found to be postsynaptic in axoaxonal synapses. Most of the C terminals degenerate following rhizotomy or are labelled following injection of the parent dorsal root ganglia with tritiated amino acids. Approximately one-fifth of the R profiles are derived from dorsal roots. F profiles do not appear to be of dorsal root origin in any case. It is concluded that neurofilamentous alterations represent the degeneration of larger-diameter (Aβ) axons which distribute to the deeper dorsal horn and that electron-lucent degeneration represents the termination of Aδ fibers. Electron-dense degeneration thought to represent the termination of nonmyelinated axons (C fibers) in the superficial dorsal horn is not seen in the deeper dorsal horn and it is concluded that C fibers do not project to the deeper laminae.     

Spinal cord projections from hindlimb muscle nerves in the rat studied by transganglionic transport of horseradish peroxidase, wheat germ agglutinin conjugated horseradish peroxidase, or horseradish peroxidase with dimethylsulfoxide

The spinal cord projections of four different groups of hindlimb muscle nerve branches--the medial and lateral gastrocnemius nerves, muscle branches of the deep peroneal nerve, muscle branches of the femoral nerve, and a nerve to the hamstring muscles--were studied with transganglionic transport of horseradish peroxidase (HRP) in the rat. The influence of varying the postoperative survival (3, 6, and 10 days) and of using wheat germ agglutinin-HRP conjugate (WGA-HRP), or HRP with dimethylsulfoxide (DMSO) instead of free HRP was studied for the gastrocnemius nerves. After 3 days' survival following application of HRP to the gastrocnemius nerves, fine granular labeling was found mainly in lamina V in L4-5, and coarse granular labeling was found in Clarke's column as far caudally as L2, and in laminae VI and VII predominantly in Th12-L2. After 6 or 10 days' survival, the fine labeling in lamina V was sparse or absent, whereas the coarse labeling appeared to remain or to be only slightly reduced in Clarke's column and in laminae VI and VII. No labeling suggestive of terminals was observed in laminae I-III from the gastrocnemius nerves. Except for sparse labeling in lamina I in some of the cases and some minor differences rostrocaudally, the spinal distribution of labeling was similar to that from the other nerves investigated. The distribution of labeling obtained after application of WGA-HRP or HRP with DMSO to the gastrocnemius nerves was very similar to that obtained with free HRP after 3 days' survival. The results indicate that the spinal cord projections of hindlimb muscle nerves in the rat distribute mainly in the deep part of the dorsal horn and in the intermediate zone. Furthermore, the lack of labeling suggestive of terminals in laminae I-III from the gastrocnemius nerves suggests, in conflict with earlier findings in the cat, that primary afferent fibers from muscles do not necessarily terminate in these laminae in the rat. The results suggest, furthermore, that fine granular labeling found in lamina V represents fine-calibered afferent fibers. Finally, the similar spinal projection patterns of the different muscle nerves investigated suggest either a less developed or an essentially different somatotopic organization for muscle afferents compared to cutaneous afferents, as revealed in earlier studies.     

Cortical projections to superficial laminae of the dorsal horn and to the ventral horn of the spinal cord in the North American opossum. Studies using the orthograde transport of WGA-HRP

The orthograde transport of horseradish peroxidase conjugated to wheat germ agglutinin (WGA-HRP) has been used to study the distribution of corticospinal axons in adult and pouch-young opossums. As predicted from the results of degeneration and autoradiographic experiments, injections of WGA-HRP into limb areas of somatic motor-sensory cortex labeled axons in the dorsal and lateral funiculi of the cervical and upper thoracic spinal cord which could be traced to dense terminal zones in laminae III–VI. In addition, we obtained evidence for the presence of a few cortical axons in the ventral white matter and for innervation of the medial part of laminae I and II, laminae VII and VIII and lamina X. A few cortical axons are even present in lamina IX.     

Spinocerebellar projections to lobules III to V of the anterior lobe in the cat,as studied by retrograde transport of horseradish peroxidase

Spinocerebellar tract (SCT) neurons projecting to lobules III to V of the cerebellar anterior lobe were identified by the retrograde horseradish peroxidase technique. SCT neurons projecting to lobule III with crossed ascending axons were located mainly in the central cervical nucleus (CCN), the medial part of lamina VII of L6 to the caudal segments, and the dorsal horn (lamina V) and ventral horn (lamina VIII) of the sacral-caudal segments. Spinal border cells with crossed ascending axons also projected to lobule III. SCT neurons projecting to this lobule with uncrossed ascending axons were located in the medial part of lamina VI of the cervical segments and the middle part of lamina VII of C6 to T1, lamina V of the lower cervical, thoracic and the lumbar segments, Clarke's column including marginal neurons, and the medial part of lamina VI of L5 and L6. These neuronal groups also projected to lobule IV, except for those present caudal to L6 (in the medial part of lamina VII, and laminae V and VIII of the sacral-caudal segments). A far smaller number of similar neurons projected to lobule V. Injections of HRP restricted to the vermal region labeled mainly neurons in the CCN and Clarke's column while restricted injections to the intermediate-lateral regions labeled ipsilaterally spinal border cells, lamina V neurons, and Clarke column neurons, especially of the lumbar segments as well as marginal neurons of this column.     

Immunocytochemical localization of glutamate decarboxylase in rat spinal cord.

The GABA synthesizing enzyme, glutamate decarboxylase (GAD), has been localized by light and electron microscopy in the rat lumbosacral spinal cord using a peroxidase-labeling antibody technique. The light microscopic localization shows heavy, punctate reaction product for GAD in the dorsal horn laminae I-III. Moderately heavy reaction product is also seen in the deeper dorsal horn laminae IV-VI, the medial aspect of the intermediate gray (lamina VII) and the region around the central canal (lamina X). A moderately light concentration of GAD reaction product is observed in the ventral horn, and punctate deposits of reaction product also are seen on motoneuron cell bodies. The punctate distribution of reaction product for GAD in both ventral and dorsal horns, as visualized by light microscopy, corresponds to GAD-containing synaptic terminals seen by electron microscopy in comparable regions of the spinal gray. Many more GAD-positive terminals are observed in dorsal horn laminae I-III than in deeper laminae IV-VI. GAD-containing terminals in the dorsal horn are presynpatic to dendrites and cell bodies. Gad-containing terminals presynaptic to other axon terminals are observed also, and they are more numerous in laminae II and III. In the ventral horn motor nuclei, GAD-positive knobs are presynaptic to large and small dendrites and motoneuror cell bodies. In addition, small GAD-containing terminals also are presynaptic to larger axonal terminals which are in turn presynaptic to motoneuron somata. The observation of GAD-containing terminals presynaptic to dendrites and cell bodies in both dorsal and ventral horns is compatible with the evidence suggesting that GABA terminals may mediate postsynaptic inhibition of spinal interneurons and motoneurons. The additional finding of GAD-positive terminals presynaptic to other axonal terminals in the dorsal horn and motor nuclei is consistent with the growing evidence that GABA also may be the transmises mediating presynaptic inhibition via axo-axond synapses in the spinal cord.     

Fine afferent fibers from viscera do not terminate in the substantia gelatinosa of the thoracic spinal cord

Transganglionic transport of horseradish peroxidase (HRP) has been used to study the anatomy of the central projection of somatic and visceral afferent fibers to the thoracic spinal cord of the cat. A dense concentration of somatic afferent fibers and terminals was found in laminae I and II of the dorsal horn and more scattered terminals were present in laminae III, IV and V and in Clarke's column. In contrast, visceral afferent fibers and terminals were found only in lamina I or reaching lamina V via a small bundle of fibers located in the lateral border of the dorsal horn. These results indicate that fine afferent fibers from viscera, unlike those of cutaneous origin, do not project to the substantia gelatinosa (lamina II) of the dorsal horn.     

Spinocerebellar projections in the pigeon with special reference to the neck region of the body

Neck sensory information is important for control of head and body movements in all vertebrates. Neuroanatomic tracing methods were used to study the pathways of neck afferent systems. Both the projection of primary afferent fibers and of secondary afferent pathways to brainstem and cerebellum were investigated with the anterograde transport of dextran amines as tracers (biotinylated dextran amine and tetramethyl rhodamine dextran amine). For comparison, the projections of spinocerebellar systems of wing and leg were studied also. Complementary experiments using retrograde tracers (Fast Blue, tetramethyl rhodamine dextran amine, rhodamine isothiocyanate) injected into the cerebellum served to corroborate the results of the anterograde tracing experiments. Primary neck afferent fibers terminated in the spinal gray substance with dense terminal fields in laminae I to V of the dorsal horn and lamina IX of the ventral horn as well as in the marginal nuclei located at the lateral border of the spinal cord. In the brainstem, dense terminal fields were seen in deep layers of the medullary dorsal horn, in the external cuneate nucleus, and in group x. Secondary neck afferents arising from ventral horn cells showed a significant projection to the descending and medial vestibular nuclei and to the medial cerebellar nucleus. Terminals were found both in the anterior and the posterior cerebellum. A quantitative evaluation disclosed that most terminals of neck afferents distributed in lobules II-IV of the anterior cerebellum and lobule IX of the posterior cerebellum. With injections aimed at spinocerebellar neurons located into the cervical and lumbosacral enlargements, no projections were found in the vestibular or deep cerebellar nuclei. Projections from the cervical enlargement were concentrated in lobules III-V and those from the lumbosacral enlargement in lobules III-VI. This points to a rostrocaudal somatotopic representation of neck, wing, and leg in the anterior cerebellum. The results of the retrograde tracing experiments support such a somatotopic organization.     

Trajectory of group Ia afferent fibers stained with horseradish peroxidase in the lumbosacral spinal cord of the cat: three dimensional reconstructions from serial sections.

A reconstruction was made of the intramedullary trajectory of 23 physiologically identified Ia afferents from cat hind limb muscles (medial gastrocnemius, soleus, plantaris, flexor digitorum-hallucis longus, and hamstring). The afferents were stained by intra-axonally injected HRP. The axons of these afferents were traced over distances of 5.8 mm to 15.7 mm rostrocaudally. In the dorsal funiculus fibers from all the muscles showed a similar course and similarly bifurcated into an ascending and a descending branch. The mean diameters of stem axons, ascending branches, and descending branches were 6.6 micrometer, 5.8 micrometer, and 3.0 micrometer, respectively. Within the analyzed lengths of the spinal cord five to eleven collaterals were given off from the two branches. The distances between adjacent collaterals of the ascending and descending branches averaged 1200 micrometer and 790 micrometer, respectively. The collaterals as a rule passed through the medial half of the dorsal horn before they entered the deeper parts of the gray matter. The terminal distribution areas common to all Ia collaterals were: (1) the medial half of the base of the dorsal horn, mainly lamina VI: (2) lamina VII; and (3) lamina IX. The numbers of terminals were largest in lamina IX and smallest in lamina VII. The density of terminals in lamina IX was highest in the homonymous motor cell column. The terminal distribution areas of adjacent collaterals showed no overlap in the sagittal plane. Terminal branches carried one bouton terminal and up to six boutons en passage with an average of 1.8 terminals per terminal branch. Apparent axosomatic and axodendritic contacts were seen on small-sized and medium-sized neurons in laminae V-VI, medium-sized neurons in lamina VII, and large neurons in lamina IX. One motoneurons was contacted by an average of 3.3 terminals. In addition to the common features, Ia collaterals of various muscles of origin showed some differences in their trajectories in the ventral horn, and in their terminations in the gray matter.     

Localization of the Serotonin Transporter in Rat Spinal Cord

The spinal cord is richly innervated by serotoninergic fibres originating from the raphe nuclei. The localization of the terminating component of serotoninergic neurotransmission, the serotonin transporter SERT1, was found in both the dorsal and ventral horns, especially at the level of the cervical and lumbar segments. Within the thoracic region, we observed a heavily labelled bundle in the intermediolateral nucleus of lamina VII. A low density of stained fibres was encountered in the sacral spinal cord. In contrast to homogeneous staining of motor nuclei, a differential labelling of laminae was seen in the dorsal horn, with laminae I, III and IV exhibiting a higher density of immunopositive terminals than the medial part of lamina II. High magnification revealed a preferential accumulation of serotonin transporter staining within nerve endings and varicosities of thin fibres. Double immunofluorescence staining demonstrated a co-localization of serotonin and its uptake system within these varicosities. These results show that the serotonin transporter is highly expressed in the rat spinal cord and that its distribution parallels the serotoninergic innervation. They also reinforce the view that varicosities are important neuronal structures, which modulate the function of dorsal and ventral horn neurons by releasing serotonin.