Saburo Kawaguchi

Runx3 controls the axonal projection of proprioceptive dorsal root ganglion neurons

Dorsal root ganglion (DRG) neurons specifically project axons to central and peripheral targets according to their sensory modality. The Runt-related genes Runx1 and Runx3 are expressed in DRG neuronal subpopulations, suggesting that they may regulate the trajectories of specific axons. Here we report that Runx3-deficient (Runx3−/−) mice displayed severe motor discoordination and that few DRG neurons synthesized the proprioceptive neuronal marker parvalbumin. Proprioceptive afferent axons failed to project to their targets in the spinal cord as well as those in the muscle. NT-3-responsive Runx3−/− DRG neurons showed less neurite outgrowth in vitro. However, we found no changes in the fate specification of Runx3−/− DRG neurons or in the number of DRG neurons that expressed trkC. Our data demonstrate that Runx3 is critical in regulating the axonal projections of a specific subpopulation of DRG neurons.

Electrophysiological studies on the cerebellocerebral projections in monkeys.

Summary1.Responses evoked by stimulation of the cerebellar and thalamic nuclei were recorded by microelectrodes introduced at various depths in the cerebral cortex of monkeys (Macaca mulatta) under light Nembutal anaesthesia.2.Stimulation of the medial (fastigial) cerebellar nucleus produced, at a latency of 4–5 msec, deep thalamo-cortical (T-C) responses (surface positivedeep negative potentials) mainly in the medial part of the precentral gyrus (area 4, “motor area for hindlimb”) and in the superior parietal gyrus (area 5) on both contralateral and ipsilateral sides to the nucleus stimulated.3.Stimulation of the lateral (dentate) cerebellar nucleus elicited, at a latency of about 3 msec, superficial T-C responses (surface negative-deep positive potentials) predominantly in the lateral part of the precentral gyrus (area 4, “motor area for forelimb and face”) and in the rostromedial part of the gyrus (area 6, premotor area) on the contralateral side.4.Stimulation of the interpositus cerebellar nucleus set up superficial T-C responses chiefly in the motor area between those influenced by the medial and the lateral cerebellar nucleus stimulation and also in the premotor area on the contralateral side.5.The respective areas responsive to the medial, interpositus and lateral nucleus stimulation overlapped considerably each other in the motor cortex.6.Comparison of the responses in the cortex induced by stimulation of the cerebellar and thalamic nuclei indicated different relay portions in and around the VA-VL region of the thalamus for the superficial and the deep T-C responses respectively.7.Functional implications of the results were discussed in referring to the cerebellocerebral projections in cats.

Electrophysiological studies on cerebello-cerebral projections in the cat

Summary1.Cerebello-cerebral projections were electrophysiologically investigated in cats under light Nembutal anaesthesia. Marked responses were produced by stimulation of the interpositus and the lateral nucleus of the cerebellum not only in the pericruciate but also in the suprasylvian cortical areas, both areas being contralateral to the cerebellar nuclei stimulated. Medial nucleus stimulation set up little or no response in the cerebral cortex.2.The previous electrophysiological study on thalamo-cortical (T-C) projections showed two different kinds of responses in the cortex due presumably to two different T-C projection systems, i. e., deep and superficial T-C responses (see Sasaki et al., 1970). According to laminar field potential analysis, the response in the pericruciate area is characterized by a deep T-C response which is often followed by a superficial T-C response, whereas the response in the parietal cortex consists of a pure superficial T-C response. Intracellular potential changes in cortical neurones elicited by cerebellar nucleus stimulation were consistent with the results of laminar field potential analysis.3.Comparison between laminar field potentials in the same cortex produced by thalamic and cerebellar nucleus stimulation suggests that the response in the pericruciate cortex is mediated by the ventral lateral nucleus and that the response in the parietal cortex is relayed by the ventral anterior nucleus of the thalamus.

Survival of Neural Stem Cells in the Cochlea

Adult rat hippocampus-derived neural stem cells (NSCs) have been reported to have been successfully grafted in several brain regions. To evaluate the possibility of treatment of sensorineural hearing loss using NSCs, survival of NSCs in the cochlea was estimated. NSCs were grafted into newborn rat cochleas. Within 2-4 weeks of grafting to the cochlea, some NSCs survived in the cochlear cavity. Some of them had adopted the morphologies and positions of hair cells. This suggests that NSCs can adapt to the environment of the cochlea and gives hope for treatment of the damaged cochlea and sensorineural hearing loss.Adult rat hippocampus-derived neural stem cells (NSCs) have been reported to have been successfully grafted in several brain regions. To evaluate the possibility of treatment of sensorineural hearing loss using NSCs, survival of NSCs in the cochlea was estimated. NSCs were grafted into newborn rat cochleas. Within 2-4 weeks of grafting to the cochlea, some NSCs survived in the cochlear cavity. Some of them had adopted the morphologies and positions of hair cells. This suggests that NSCs can adapt to the environment of the cochlea and gives hope for treatment of the damaged cochlea and sensorineural hearing loss.

On the cerebello-thalamo-cerebral pathway for the parietal cortex.

Summary1.The cerebello-thalamo-cerebral projection system mediating the cerebellar-induced “superficial thalamo-cortical (T-C) response” (the basic type of the so-called recruiting response) to the anterior part of the middle suprasylvian gyrus was investigated electrophysiologically. Responses of thalamic neurones to stimulation of the cerebral cortex and the cerebellar nucleus (medial, interpositus and lateral) were recorded by microelectrodes.2.In the anterior portions of the ventral thalamic nuclear complex, presumably in and/or around the ventral anterior (VA) nucleus, there were found neurones responding antidromically to stimulation of the suprasylvian cortex and orthodromically to that of the interpositus and the lateral nucleus of the cerebellum. They were called P neurones. The neurones responding antidromically to stimulation of the anterior sigmoid cortex and orthodromically to that of the cerebellar nuclei located mostly caudo ventrolateral to the place of P neurones, presumably in and/or around the ventral lateral (VL) nucleus. These were called F neurones.3.The cerebellar excitation of P neurones was estimated on its latency to be monosynaptic and was usually followed by an inhibition lasting for more than 100 msec. Large unitary EPSPs were sometimes noted in P neurones on cerebellar stimulation as well as spontaneously. It was concluded that P neurones constitute the direct T-C projection system mediating the superficial T-C response (e. g., recruiting response) to the parietal cortex.

Tenascin-C regulates proliferation and migration of cultured astrocytes in a scratch wound assay

Tenascin-C (TNC), an extracellular matrix glycoprotein, is involved in tissue morphogenesis like embryogenesis, wound healing or tumorigenesis. Astrocytes are known to play major roles in wound healing in the CNS. To elucidate the roles of TNC in wound closure by astrocytes, we have examined the morphological changes of cultured astrocytes in a scratch wound assay and measured the content of soluble TNC released into the medium. We have also localized the expression of TNC mRNA, TNC, glial fibrillary acidic protein (GFAP), vimentin and integrin beta1. After wounding, glial cells rapidly released the largest TNC isoform and proliferated in the border zones. Subsequently, they became polarized with unidirectional processes and finally migrated toward the denuded area. The proliferating border zone cells and pre-migratory cells intensely expressed TNC mRNA, TNC-, vimentin-, GFAP- and integrin beta1-like immunoreactivity, while the migratory cells showed generally reduced expression except the front. Exogenous TNC enhanced cell proliferation and migration, while functional blocking with anti-TNC or anti-integrin beta1 antibody reduced both of them. These results suggest that mechanical injury induces boundary astrocytes to produce and release TNC that promotes cell proliferation and migration via integrin beta1 in an autocrine/paracrine fashion.

Grafting of choroid plexus ependymal cells promotes the growth of regenerating axons in the dorsal funiculus of rat spinal cord: a preliminary report.

Nerve regeneration in the central nervous system has been studied by grafting various tissues and cells. In the present study, we demonstrated that choroid plexus ependymal cells can promote nerve regeneration when grafted into spinal cord lesions. The choroid plexus was excised from the fourth ventricle of adult rats (Wistar), minced into small fragments, and grafted into the dorsal funiculus at the C2 level in adult rat spinal cord from the same strain. Electron microscopy and fluorescence histochemistry showed that ependymal cells of the grafted choroid plexus intimately interacted with growing axons, serving to support the massive growth of regenerating axons. CGRP-positive fibers closely interacted with grafted ependymal cells. HRP injection at the sciatic nerve showed that numerous HRP-labeled regenerating fibers from the fasciculus gracilis extended into the graft 7 days after grafting. This regenerating axons from the fasciculus gracilis was maintained for at least 10 months, with some axons elongating rostrally into the dorsal funiculus. Evoked potentials of long duration were recorded at a level ca. 5 mm rostral to the lesion in the rats 8 to 10 months after grafting. These findings indicate that choroid plexus ependymal cells have the ability to facilitate axonal growth in vivo, suggesting that they may be a promising candidate as graft for the promotion of nerve regeneration in the spinal cord.

Mossy fibre and climbing fibre responses produced in the cerebellar cortex by stimulation of the cerebral cortex in monkeys

Summary1.Responses in the cerebellar cortex induced by stimulation of several areas of the cerebral cortex were recorded and identified electro-physiologically to be due to mossy fibre and climbing fibre volleys, and their distributions were explored in the anterior and the posterior lobes of the cerebellum in monkeys. Early mossy and late climbing fibre responses at latencies of 4–5 and 15–18 msec respectively were recorded in certain areas of the cerebellar cortex. They were usually predominant on the contralateral side to the stimulation.2.Stimulation of the lateral part of the motor cortex (forelimb and face area) evoked mossy and climbing fibre responses mainly in the ansoparamedian lobules and in the caudal part of the anterior lobe (including lobulus simplex) of the cerebellar cortex, stimulation of the medial part of the motor cortex (hindlimb area) provoked the responses predominantly in the rostral part of the anterior lobe, and that of the intermediate part (areas for trunk and proximal parts of the extremities) induced the responses preponderantly in the middle part of the anterior lobe of the cerebellum.3.Stimulation of the parietal association cortex (area 5) elicited mossy and climbing fibre responses chiefly in the anterior lobe of the cerebellum. The premotor cortex innervates wide areas of the anterior and posterior lobes on both contralateral and ipsilateral sides. The frontal association cortex showed the projections on even wider areas of the cerebellar cortex, although the responses were relatively small in size.4.The results were compared with those obtained in cats and considered in referring to the cerebro-cerebellar loops in monkeys.

Postnatal development of the geniculocortical projection in the cat: Electrophysiological and morphological studies

SummaryUsing laminar field potential analysis, we examined responses elicited by both photic and optic nerve stimulations in 30 kittens of 0–65 days of age and in three adult cats. In adult cats, the response in the visual cortex on optic nerve stimulation is a wave complex which consists mainly of surface positive-depth negative (sP-dN) potentials. By contrast, the response in neonates consists of two surface negative — depth positive (sN-dP) waves. In 2 weeks, preceding the sN-dP waves, an sP-dN wave appears. As age increases, the sP-dN wave becomes of higher voltage and the sN-dP waves become of lower voltage. Thus, the configuration of the response resembles that of adult cats in 3–4 weeks. Both photic and optic nerve stimulations elicit responses of the same configuration in the same area. The extent of the responsive area is exactly the same at any age as in adult cats.Using the orthograde HRP method, we examined terminals of the geniculocortical afferent in 23 kittens of 0–43 days of age. The density of labeled terminals in layer I is much higher in kittens before 1 week of age (n = 8) than in kittens after 1 month of age (n = 5), whereas the density of labeled terminals in layer IV is higher in the older kittens than in the younger kittens. These electrophysiological and morphological changes are correlated in reference to the maturation of the neuronal circuit in the visual cortex.

Spontaneous regeneration of the pyramidal tract after transection in young rats

Spontaneous regeneration of the pyramidal tract after transection of the medullary pyramid was examined in young rats by the anterograde tracing method with wheat germ agglutinin-conjugated horseradish peroxidase. Care was taken to cut the tract as sharply as possible to minimize traumatic injuries. A very sharp cut produced edema-free lesions without subsequent formation of either cysts or scars, whereas a relatively blunt cut produced edema and later scars and/or cysts in the lesion. Regenerated projections in the latter cases were sparse, short, dispersed and largely aberrant as described in previous reports. By contrast, regenerated projections in the former cases were very much similar to normal in various respects: the amount, extension, path, formation of a compact bundle and termination. There was, however, a decisive difference from normal, that is, the additional aberrant projections.