Astrocytes and guidance of outgrowing corticospinal tract axons in the rat. An immunocytochemical study using anti-vimentin and anti-glial fibrillary acidic protein

In the present investigation the role of astrocytes and their precursors in guidance of outgrowing corticospinal tract axons in the rat is studied. Antibodies against glial fibrillary acidic protein and vimentin are used to analyse immunogen expression of glial cells, whereas the postnatal outgrowth of corticospinal tract axons through the spinal cord was studied using anterogradely transported horseradish peroxidase. The first, leading corticospinal tract axons, being the objective of the present study, are characterized by dilatations at their distal ends, the growth cones. Growth cones of pioneer corticospinal tract axons are randomly distributed in the presumptive corticospinal tract area of the ventral most part of the dorsal funiculus. A dramatic change in glial cell labelling is found from the majority being vimentin immunoreactive and glial fibrillary acidic protein-negative at birth to almost all being the reverse at the end of the fourth postnatal week. From double labelling experiments it can be concluded that the vimentin-glial fibrillary acidic protein transition occurs within astrocyte precursor cells. The absence of glial fibrillary acidic protein-immunoreactive glial cells during the outgrowth period of pioneer corticospinal tract axons indicates that they cannot play a role in the guidance of outgrowing corticospinal tract pioneer axons. Vimentin-immunoreactive glial cells are present throughout the presumptive corticospinal tract area at the time of arrival of the leading corticospinal tract fibres. The vimentin-immunoreactive glial cells, which themselves are orientated perpendicular to the outgrowing corticospinal tract axons, are mainly arranged in longitudinal tiers parallel to the rostrocaudal axis. Electron microscopically, growth cones of pioneer corticospinal tract axons frequently exhibit protrusions into vimentin-immunoreactive glial cell processes, suggesting an adhesive type of contact. Therefore, in addition to a positional role, vimentin-immunoreactive glial cells probably play a chemical role in guidance of pioneer corticospinal tract axons. A prominent vimentin-immunoreactive glial septum was noted during corticospinal tract outgrowth in the midline raphe of the medulla oblongata and spinal cord whereas it is absent in the decussation area of corticospinal tract fibres. After the first postnatal week the major vimentin-immunoreactive glial barrier either completely disappears (medullary levels) or gradually reduces to a minor glial fibrillary acidic protein-immunoreactive one (spinal cord levels).(ABSTRACT TRUNCATED AT 400 WORDS)     

The regional distribution of myelin oligodendrocyte glycoprotein (MOG) in the developing rat CNS: an in vivo immunohistochemical study

Using an immunohistochemical approach we have characterized the in vivo developmental distribution of myelin oligodendrocyte glycoprotein within the rat CNS. Myelin oligodendrocyte glycoprotein expression emerged in a non-uniform manner during the first 3 postnatal weeks. Although it was absent throughout the CNS of the newborn rat at postnatal day 0(P0), it had appeared in the spinal cord and brainstem by P7. The forebrain and cerebellum remained devoid of immunoreactivity until after P14. Myelin oligodendrocyte glycoprotein emerged at different times within the closely associated fasciculi of the dorsal funiculus. It appeared in the fasciculus cuneatus during the first postnatal week and in the fasciculus gracilis and corticospinal tracts during weeks 2 and 3 respectively. Myelin oligodendrocyte glycoprotein expression developed along a caudo-rostral gradient from spinal cord to forebrain and along an antero-posterior gradient within the CNS in general. The relationship between the onset of myelin oligodendrocyte glycoprotein expression and myelinogenesis was also investigated. In most regions, myelin oligodendrocyte glycoprotein expression lagged behind the initial appearance of myelin basic protein and Luxol Fast Blue-stained myelin by at least 1 week. These observations support the idea that myelin oligodendrocyte glycoprotein is the latest myelin protein to appear in development, only being expressed during the final stages of oligodendrocyte differentiation. Furthermore, the pattern of staggered expression within the dorsal columns indicates that localized, region-specific interactions may comprise a key element in the control of the terminal phases of oligodendrocyte differentiation.     

Postnatal development of the corticospinal tract in the rat

Summary Horseradish-peroxidase was used to anterogradely label and thus to trace the growth of corticospinal axons in rats ranging in age from one day to six months. Three to eight HRP-gels were implanted in the left cerebral hemisphere of the cortex. In each spinal cord three levels were studied, the cervical intumescence (C5), the mid-thoracic region (T5) and the lumbar enlargement (L3). The methodology employed for the electron microscopic visualization of HRP has been described previously (Joosten et al. 1987a).The outgrowth of labelled unmyelinated corticospinal tract axons in the rat spinal cord primarily occurs during the first ten postnatal days. The outgrowth of the main weve of these fibres is preceded by a number of pathfinding axons, characterized by dilatations at their distal ends, the growth cones. By contrast, later appearing unmyelinated axons, which presumably grow along the pathfinding axons, do not exhibit such growth cones. The first labelled pioneer axons can be observed in the cervical intumescence at postnatal day one (P1), in the mid-thoracic region at day three (P3) and in the lumbar enlargement at day five (P5).Prior to the entrance of the axons, the prospective corticospinal area or the pre-arrival zone is composed of fascicles consisting of unlabelled, unmyelinated fibres surrounded by lucent amorphous structures. During the outgrowth phase of the corticospinal fibres some myelinated axons could be observed within the outgrowth area even before day 14. These axons, however, were never labelled. These findings strongly suggest that the outgrowth area, which is generally denoted as the pyramidal tract, contains other axons besides the corticospinal fibres (and glial cells). The process of myelination of the labelled corticospinal tract axons in the rat spinal cord starts rostrally (C5) at about day 14 and progresses caudally during the third and fourth postnatal weeks. Although myelination seems to be largely complete at day 28 at all three spinal cord levels, some labelled unmyelinated axons are still present in the adult stage.     

Region-specific appearance of myelin constituents in the developing rat spinal cord

Summary The appearance of myelin-specific glycolipids and of myelin basic protein was studied with regard to the detailed anatomy of the rat cervical spinal cord. The expression of these constituents in particular fibre tracts and regions occurs at specific times of development between postnatal days 1 and 14. This mosaic-like appearance started in the ventral funiculus (day 1) followed by fasciculus cuneatus and ventro-lateral funiculus (day 2), and fasciculus gracilis and dorso-lateral funiculus (days 3 to 4). Cortico-spinal tract (day 11), Lissauer tract (day 14) and the commissures started to acquire myelin very late. In the grey matter, myelin constituents appeared around days 11 to 14 in a patchy pattern. These results support a concept of highly local interactions regulating oligodendrocyte differentiation. In addition, a general rostro-caudal gradient of myelin development exists in the spinal cord, which is independent of the ascending or descending nature of the fibre tracts. Appearance of myelin constitutents in the caudal spinal cord was not prevented by a neonatal transection at mid-thoracic levels.     

Growth and target finding by axons of the corticospinal tract in prenatal and postnatal rats

The growth of the corticospinal tract was studied in prenatal and neonatal rats using the anterograde transport of horseradish peroxidase injected into the cerebral cortex as a marker in lightmicroscopic preparations. The findings were compared with electron-microscopic observations on normal material at the same ages. Labelled corticofugal axons traverse the diencephalon by gestational day 17.5, reach the pontine nuclei by gestational day 19.5, and the caudal limit of the medulla oblongata by gestational day 20.5, just before birth. On the day after birth, labelled corticospinal axons have crossed in the pyramidal decussation and extended into the dorsal columns of the upper cervical spinal cord. Corticospinal axons reach the thoracic segments by postnatal day 3, the lumbar segments by day 6 and the sacral segments by day 9. The lower end of the spinal cord is reached only after postnatal day 14. Beside the principal corticospinal tract in the dorsal columns, two other smaller corticospinal tracts occupy an intermediate position in the base of the cervical dorsal horn and a lateral position in the lateral white column. The intermediate tract is not found below cervical levels. Growth cones are seen at the tips of axons in light- and electron-microscopic material. The first corticospinal axons, less than 0.5 microns in diameter and grouped in tight fascicles, grow through a dense fabric of astrocytic and other glial processes in which no obvious pre-existing channels could be identified. Growth of corticospinal axons into the dorsal horn adjacent to the main tract is delayed until 2–3 days after the initial arrival of the tract at a given segment. This begins in the cervical segments only after the thalamocortical fibers have invaded the sensory-motor cortex though the parent pyramidal cells of the tract are still highly immature. The rate of extension of corticospinal axons is not constant. Growth down the dorsal columns is characterized by accelerated growth spurts on postnatal days 4 and 9. Much slower growth characterizes initial outgrowth through the diencephalon and later ingrowth into the spinal gray matter. There is approximately a three-fold increase in the numbers of corticospinal axons in the cervical segments between postnatal days 5 and 10. Myelination commences between postnatal days 10 and 12.It is concluded that the development of the corticospinal tract in the rat displays features that are common to other developing pathways in the rat and other species. Initial outgrowth of corticospinal axons is independent of afferent innervation, occurring at a time when the parent cell bodies are very immature. The early growth of corticospinal axons to the vicinity of their targets is followed by a substantial waiting period, comparable to that seen in other systems, before final invasion of the target. The factors responsible for the initiation of the second growth spurt, carrying axons into the target gray matter are not known. However, the final invasion of gray matter takes place only after the cells of origin of corticospinal axons have received a substantial afferent input. The rate of initial growth of corticospinal axons down the dorsal columns is not constant, but varies from region to region. Electronmicroscopy has failed to detect any morphological evidence for factors that might guide or promote the growth of corticospinal axons. The majority of corticospinal axons exclusive of the first ‘pathfinders’ seem to grow as tight fascicles in which individual axons contact only one another.     

Development of the corticospinal tract in Semaphorin3A- and CD24-deficient mice

Mutations in the gene encoding the neural recognition molecule L1 result in hypoplasia of the corticospinal tract and path finding errors of corticospinal axons at the pyramidal decussation. Candidate molecules that have been implicated in L1-dependent guidance of corticospinal axons from the ventral medullary pyramids to the contralateral dorsal columns of the cervical spinal cord include Semaphorin3A and CD24. In the present study, we anterogradely labeled corticospinal axons from the sensorimotor cortex of young postnatal Semaphorin3A- and CD24-deficient mice to elucidate potential functions of both proteins during formation of this long axon projection. Our results indicate that elongation, collateralization, fasciculation and path finding of corticospinal axons in mice proceed normally in the absence of Semaphorin3A or CD24.     

GAP-43 expression in the developing rat lumbar spinal cord.

The expression of the growth-associated protein GAP-43, detected by immunocytochemistry, has been studied in the developing rat lumbar spinal cord over the period E11 (embryonic day 11), when GAP-43 first appears in the spinal cord, to P29 (postnatal day 29) by which time very little remains. Early GAP-43 expression in the fetal cord (E11-14) is restricted to dorsal root ganglia, motoneurons, dorsal and ventral roots and laterally positioned and contralateral projection neurons and axons. Most of the gray matter is free of stain. The intensity of GAP-43 staining increases markedly as axonal growth increases, allowing clear visualization of the developmental pathways taken by different groups of axons. Later in fetal life (E14-19), as these axons find their targets and new pathways begin to grow, the pattern of GAP-43 expression changes. During the period, GAP-43 staining in dorsal root ganglia, motoneurons, and dorsal and ventral roots decreases, whereas axons within the gray matter begin to express the protein and staining in white matter tracts increases. At E17-P2 there is intense GAP-43 labelling of dorsal horn neurons with axons projecting into the dorsolateral funiculus and GAP-43 is also expressed in axon collaterals growing into the gray matter from lateral and ventral white matter tracts. At E19-P2, GAP-43 is concentrated in axons of substantia gelatinosa. Overall levels decline in the postnatal period, except for late GAP-43 expression in the corticospinal tract, and by P29 only this tract remains stained.     

Primary demyelination and regeneration of ascending axons in the dorsal funiculus of the rat spinal cord following photochemically induced injury

Summary The extent of primary demyelination and regeneration of ascending axons in the dorsal funiculus of the rat spinal cord was investigated following photochemically-induced ischaemic injury. Groups of rats were killed at intervals from 48h to 1 month after injury and a combination of light and electron microscopy and counting of axons in specific sites was used to study the axonal changes. Unmyelinated axons were noted in the dorsal rim of the lesion at its centre and at the centre of the gracile fasciculus at the caudal end of the lesion 7 days after injury. By 1 month, axons in these sites were thinly myelinated by Schwann cells or oligodendrocytes. In order to differentiate between remyelination of demyelinated axons and myelination of regenerated axons, axon counts were performed. The number of sub-pial axons present at the lesion centre did not change significantly from 48h to 1 month after injury, whereas the number of axons at the caudal end of the lesion increased significantly from 4 to 10 days after injury. We therefore conclude that sub-pial axons at the lesion centre are demyelinated between 4 and 7 days after injury and subsequently remyelinated by Schwann cells. At the caudal end of the lesion, a specific population of small diameter axons located at the centre of the gracile fasciculus regenerates for a distance of approximately 1 mm between 4 and 10 days after injury; these axons are then myelinated by oligodendrocytes or Schwann cells. In contrast, larger diameter axons of the gracile fasciculus do not show a regenerative response, demonstrating the variability of axonal responses to injury.     

Long term post-traumatic retrograde corticospinal degeneration in man

The spinal cord and brain of a man who died 18 years after a crush injury of lumbar segments contained some unusual lesions. There was a reduced number of myelinated axons in the corticospinal tracts as high as the fifth cervical segment. Such retrograde degeneration has been described in human pyramidal tracts only a few times. The results of reported studies of experimental retrograde degeneration have been inconsistent. The course of the fasciculus gracilis, as delineated by gliosis, was atypical, and an unusual glial nodule, possibly neoplastic, was present in the dorsal columns at C8.     

Myelin basic protein immunohistochemistry: a study of the early stages of myelination in the brainstem of the rat.

An immunohistochemical study of MBP distribution in the brainstem of neonate till 16 d old rats based on the peroxidase-antiperoxidase method is described. Axons already invested with immunoreactive sheaths were found in neonate rats in the ventral funiculus of the cervical spinal cord and in the medial longitudinal fascicle of the medulla oblongata. Fibres commencing with myelination showed a closely spaced array of varicosities in longitudinal sections which diminished gradually. A caudo-rostral decrease in density of myelinated fibres in the brainstem was found in the medial and dorsal longitudinal fascicles. In contrast to other pathways, myelination in the fibres of the corticospinal tract in the brainstem occurred in a strictly synchronized pattern. The same temporal pattern of myelination was also observed in the cervical corticospinal tract, except that a few myelinated fibres had been visible much earlier within the area of the tract. At the exit of cranial nerves, the transitorial zone from central to peripheral myelin was outlined by a decrease in immunostaining.     

Numbers and proportions of unmyelinated axons at cervical levels in the fasciculus gracilis of monkey and cat.

The present study is a quantitative analysis of the unmyelinated fiber population in the fasciculus gracilis of the second cervical segment of cat and monkey. We find that unmyelinated fibers represent 13.7% of the total fiber population in this pathway in the cat and 18.9% in the monkey (Macaca fascicularis). The existence of such large numbers of these axons suggests that there may be a sizeable ascending fine primary afferent pathway in the fasciculus gracilis in cat and monkey whose destination is presumably the dorsal column nuclei. These findings are of interest in regard to classic ideas that the afferent fibers in the dorsal columns are large myelinated fibers that convey fine discriminative information to the dorsal column nuclei.     

The topology of dermatomal projection in the medial lemniscal system

1. The topographic organization of first order afferent fibres in the lumbar, sacral and coccygeal dorsal roots, and in the fasciculus gracilis was studied in squirrel monkeys.2. At the entry zone, progressing from caudal to rostral, dorsal root filaments receive fibres from tail and hind-limb receptive fields which serially overlap and describe a spiral-shaped trajectory. The latter starts with tail, progresses post-axially towards the foot, crosses the foot from lateral to medial, and ascends the preaxial leg.3. In the fasciculus gracilis, this arrangement of fibres at the dorsal root entry zone is preserved in its entirety. It assumes the form of a fibre lamination, with the most caudal dorsal root fibres occupying a dorso-medial location; further rostral dorsal root fibres come to lie more ventrolaterally.4. Dorsum and sole of foot project in an overlapping and interdigitating manner to the fibre lamina of the 7th lumber dermatome in the fasciculus gracilis. Thereby, dorsum and sole of foot behave in the projection as if they were one and the same surface.5. The argument is presented that the foot and its projection on to the cross-sectional plane of the dorsal funiculus are topologically equivalent and that the hind-limb as a whole and its projection are not. On the other hand, homotopic mapping of the foot together with the sequential fibre organization in the dorsal funiculus enable many more types of closed curves on the body surface to remain arc-wise connected in the projection than would otherwise be possible.     

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.     

Numbers and proportions of unmyelinated axons at cervical levels in the fasciculus gracilis of monkey and cat

The present study is a quantitative analysis of the unmyelinated fiber population in the fasciculus gracilis of the second cervical segment of cat and monkey. We find that unmyelinated fibers represent 13.7% of the total fiber population in this pathway in the cat and 18.9% in the monkey (Macaca fascicularis). The existence of such large numbers of these axons suggests that there may be a sizeable ascending fine primary afferent pathway in the fasciculus gracilis in cat and monkey whose destination is presumably the dorsal column nuclei. These findings are of interest in regard to classic ideas that the afferent fibers in the dorsal columns are large myelinated fibers that convey fine discriminative information to the dorsal column nuclei.     

The central projections of visceral primary afferent neurons of the inferior mesenteric plexus and hypogastric nerve and the location of the related sensory and preganglionic sympathetic cell bodies in the rat

Summary The central projections of visceral primary afferents of the inferior mesenteric plexus and hypogastric nerve of the rat were investigated using the transganglionic transport of horseradish peroxidase (HRP). In addition, the location of the corresponding spinal ganglion cells as well as the preganglionic sympathetic neurons is demonstrated.Labelled afferent axons were found in dorsal roots, dorsal root entry zone (preferentially in its lateral part), in all parts of the tract of Lissauer, and in the dorsolateral funiculus. Preterminal axons and/or terminals were distributed mainly to laminae I, IIa and the nucleus of the dorsolateral funiculus. Fewer afferents reached laminae IIb, III–V and X. Afferent projections are densest at L1 and 2 and the caudal T13, but extend up to T10 rostrally, and at least down to L4 caudally. A few visceral afferents ascend to the nucleus gracilis.The great majority of sensory and preganglionic sympathetic cell bodies is located at levels L1 and 2 bilaterally. A few cells are found in decreasing numbers rostrally up to T11.Preganglionic sympathetic neurons (PSN) are located in nucleus intermediolateralis (IML), n. intercalatus (IC) and n. commissuralis dorsalis (DCN). Axons of DCN and IC neurons run laterally, joining those of IML neurons on their way to the ventral roots. Dendrites of IML neurons ramify in all directions but preferentially to the dorsal horn and dorsolateral funiculus. Dendrites of IC and DCN neurons are distributed mainly mediolaterally, the latter also ventrally around the canalis centralis.     

Localization of aggrecan and versican in the developing rat central nervous system.

The localization of aggrecan and mRNA splice variants of versican in the developing rat central nervous system has been examined by using specific polyclonal antibodies to the nonhomologous glycosaminoglycan attachment regions of these hyaluronan-binding chondroitin sulfate proteoglycans. At embryonic day 16 (E16), aggrecan and versican splice variants containing either or both the alpha-and beta-domains are present in the marginal zone and subplate of the cerebral cortex and in the amygdala, internal capsule, and the optic and lateral olfactory tracts. There is strong staining of versican but not of aggrecan in the hippocampus and dentate gyrus by E19, whereas both aggrecan and alpha-versican are present in the fimbria. At E19, aggrecan is seen throughout the cerebral cortex, whereas the distribution of versican is considerably more limited, being confined essentially to the marginal zone and subplate. At 1 week postnatal, both aggrecan and versican are present in the prospective white matter and in the molecular and granule cell layers of the cerebellum, but neither proteoglycan is seen in the external granule cell layer. alpha- but not beta-versican staining is seen in Purkinje cells, and aggrecan staining of Purkinje cells is also rather minimal. In the spinal cord at E13, aggrecan is present in the dorsal root entry zone, ventral funiculus, mantle layer, and floor plate, as well as in the dorsal root ganglia and ventral roots. However, alpha-versican is confined to the dorsal root entry zone and the ependyma surrounding the spinal canal, and beta-versican is not present in spinal cord parenchyma at this developmental stage, being limited to the surrounding connective tissue. By E19, there are significant amounts of all three proteoglycans in the spinal cord. Aggrecan staining is most intense in the lateral funiculus and the fasciculi gracilis and cuneatus, where alpha-versican staining is also strong. In contrast, beta-versican is seen predominantly in the motor columns. Differences in the localization and temporal expression patterns of these chondroitin sulfate proteoglycans suggest that, like neurocan and phosphacan, they have partially complementary roles during central nervous system development.     

The taiep rat: A myelin mutant with an associated oligodendrocyte microtubular defect

Summary This report describes a new inherited disorder of myelination in the rat, namedtaiep, in which failure of normal myelination of the CNS and subsequent demyelinatiori result in a progressive neurological disturbance. At two months of age, myelin is present throughout the spinal cord, but is immature in the fasciculus gracilis and corticospinal tracts despite the presence of abundant oligodendrocytes. By 12 months, myelin has largely been lost in these spinal cord tracts and also in more rostral parts of the CNS, such as the cerebellum and optic nerves. Other funiculi of the spinal cord show a more diffuse lack of myelin. Oligodendrocytes develop a unique cellular abnormality, most obviously in older rats, which is characterized by the accumulation of microtubules throughout their cytoplasm. As the mutant rats age, there is a continued protracted breakdown of myelin throughout the CNS, with evidence suggesting either persistent hypomyelination or attempts at remyelination of affected axons. It is proposed that the microtubular defect in oligodendrocytes results in a disruption of the normal myelination process in certain areas of the CNS of this mutant, and eventually leads to failure of maintenance of the myelin sheath.     

Ryk-mediated Wnt repulsion regulates posterior-directed growth of corticospinal tract

Guidance cues along the longitudinal axis of the CNS are poorly understood. Wnt proteins attract ascending somatosensory axons to project from the spinal cord to the brain. Here we show that Wnt proteins repel corticospinal tract (CST) axons in the opposite direction. Several Wnt genes were found to be expressed in the mouse spinal cord gray matter, cupping the dorsal funiculus, in an anterior-to-posterior decreasing gradient along the cervical and thoracic cord. Wnts repelled CST axons in collagen gel assays through a conserved high-affinity receptor, Ryk, which is expressed in CST axons. Neonatal spinal cord secretes diffusible repellent(s) in an anterior-posterior graded fashion, with anterior cord being stronger, and the repulsive activity was blocked by antibodies to Ryk (anti-Ryk). Intrathecal injection of anti-Ryk blocked the posterior growth of CST axons. Therefore, Wnt proteins may have a general role in anterior-posterior guidance of multiple classes of axons.     

Death of oligodendrocytes and microglial phagocytosis of myelin precede immigration of Schwann cells into the spinal cord

Small, circumscribed electrolytic lesions were made in the upper cervical corticospinal tract in adult rats. In the centre of the lesion, the axons and all other tissue elements were totally destroyed. Surrounding this region of destruction is an area of tissue which is only partially damaged. In this area TUNEL positive staining of contiguous rows of tract glial cells indicates massive oligodendrocytic apoptosis at 1–3 days after operation, but axons, astrocytes and blood vessels survive. From around 4 days, the corticospinal axons in this area are demyelinated, and the microglia contain ingested myelin, identified in electron micrographs as characteristic MBP immunoreactive laminar cytoplasmic bodies. After around 3 weeks, large numbers of Schwann cells, continuous with those on the pial surface of the spinal cord, accumulate along the lesion track and selectively infiltrate the perilesional reactive area, where they mingle intimately with the phagocytic microglia. Electron micrographs show that at this time basal lamina-enclosed Schwann cell processes establish non-myelinated ensheathment of axons. From around 4 weeks after operation, prominent Schwann cell myelination is indicated by P0 immunoreactivity, and peripheral type, one-to-one myelination in electron micrographs. Thus the effect of the selective loss of oligodendrocytes is to first activate microglia, and then to induce a replacement of myelin by Schwann cells.     

Golgi studies of the nucleus gracilis in the rat

With the use of Golgi-Kopsch and rapid Golgi techniques, seven different neuronal types were described in the nucleus gracilis of the rat. In the caudal part of the nucleus the dendrites were grouped in vertical bundles origented parallel to the collaterals of the fibers in the fasciculus gracilis. Some neurons had dendrites associated with several dendritic bundles separated in the horizontal or parasagittal planes. Two types of interneurons were found in the caudal part of the nucleus. The first had its axon confined within a dendritic column formed in part by its own dendritic arborization. The axon of the second interneuron projected to a nearby dendritic column. In the rostral part of the nucleus, the various neuronal types had dendrites oriented either transversely or along the axis of the nucleus. Fibers from the fasciculus gracilis, at rostral levels, bend to course across the nucleus sending collaterals along the parasagittal plane. Thus the dendrites of the neurons in this region were directed perpendicular and/or parallel to the afferent fibers. Afferent axons to the nucleus gracilis from the pyramidal tract and medullary reticular formation were also described.