Plasticity of GABA- and glutamate-containing terminals in the mouse thalamic ventrobasal complex deprived of vibrissal afferents: an immunogold-electron microscopic study.

GABA and glutamate immunogold staining demonstrated that nerve cells of the thalamic ventrobasal complex (VB) of mice were positive exclusively for glutamate. None of the neuronal perikarya reacted the GABA antibody. By using alternate thin sections of the normal VB, it was also shown that large "specific" somatosensory and small corticothalamic terminals, both of which contained spherical synaptic vesicles, exhibited only glutamate-like immunoreactivity. A third axonal type, containing flat-ovoid synaptic vesicles, stained only for GABA. Seventy-five days after coagulation of the vibrissal follicles in newborn mice, a characteristic multiplication of GABA positive axon terminals was observed. In addition, it was demonstrated that, similarly to modified cortical endings (Hámori et al., J. Comp. Neurol. 254:166-183, '86), many GABA positive terminals appeared as specific afferent endings, replacing the missing "specific" vibrissal afferents. This finding shows a remarkable plasticity of inhibitory GABA axons during developmental synaptogenesis and provides further evidence that the size, location, and the type of attachment of presynaptic terminals are dependent on their postsynaptic target.

An image analysis morphometric method for the study of myelinated nerve fibers from mouse trigeminal root.

For the morphometric light microscopic study of myelinated fibers in mouse trigeminal root, it was necessary to write: (1) an entirely automatic analysis program for the myelinated axons inside the myelin sheath, based on the detection of the myelin sheaths, and (2) an interactive analysis program for the myelinated fibers outside the myelin sheath, due to the high density of compactness of the myelinated fibers based on an indirect fiber individualization by reconstructing them from their axons. In the latter, a semiautomatic correction method (drawing the profile contours with a light pen) allowed compensation for the failures of the automatic method, except for the smallest fibers, which represented 8% of the total. Using these programs, 95% of the axons could be measured and 92% of the myelinated fibers whose axons were analyzed could be measured. The area-equivalent diameter was independent of the detection method; it is a correct-size measurement parameter for axons and fibers that is unrelated to their shape. The projected diameter, an estimation of the perimeter obtained by measurement of the profile projections, depended upon the detection method because the profile contour was influenced by the detection method; it thus takes into account the profile shape. For myelinated fibers, whose analysis program used two detection methods (automatic and semiautomatic), there was an average difference of 16% between the projected diameters obtained with these two methods, whereas the equivalent diameter value was the same. The fiber circularity factor could not be precisely estimated because of the detection error; the axon circularity factor was more reliable since the axon detection was completely automatic.

Immaturity of muscle fibers in the congenital form of myotonic dystrophy: its consequences and its origin.

Skeletal muscle maturation is impaired in children with congenital myotonic dystrophy. This immaturity is characterized at the light microscopy level by an abnormal presence of myotubes, small fascicles of muscle fibers, thin myofibers, and delayed muscle fiber type differentiation with a peripheral halo lacking mitochondrial oxidative enzyme activity. At an ultrastructural level, the characteristics are a paucity of myofibrils with a peripheral rim devoid of mitochondria and myofibrils in the fibers. In time the muscle is able to gain a certain degree of maturity as shown in one of our cases who had two successive muscle biopsies. The muscle, however, never becomes normal but retains discrepancies in fiber size and fiber type distribution and shows some fiber necrosis. Maturation of the motoneurons is normal, which may explain necrosis of immature muscle fibers. In an experimental study carried out to look for evidence of a circulatory factor in mothers of children with congenital myotonic dystrophy, it was found that sera from these mothers administered intra-peritoneally to newborn rats does in fact impair muscle maturation, whereas rats injected similarly with sera from control women showed normal muscle maturation.

Morphological alterations in subcortical vibrissal relays following vibrissal follicle destruction at birth in the mouse.

Morphological modifications of two subcortical vibrissal relays were analyzed, following destruction of vibrissal follicles in newborn mice. The volume of the nucleus interpolaris (NI) of the trigeminal nuclear complex in the brainstem decreased by 33%, while the number of its neuronal perikarya decreased only moderately. Vibrissal deafferentation caused no shrinkage of the ventrobasal complex (VB). In the damaged medial vibrissal part of VB (VBm), however, neuronal density was higher than normal, indicating the prevention or retardation of physiologically programmed cell death in the afferentation deprived thalamic somatosensory relay station. It is suggested that the difference in neuron density produced by deafferentation is related to the states of maturation at birth of the two subcortical vibrissal relays. Following vibrissal deafferentation the basic organization of the synaptic neuropil appeared to be similar to the control. Quantitative electron microscopic (EM) analysis revealed, however, an increased number of axon terminals with ovoid synaptic vesicles in both deafferented relay stations. The increased density of gamma-aminobutyric acid (GABA)-immunostained boutons observed in the VBm following vibrissal deprivation suggested a compensatory increase most probably of the inhibitory axon endings. Quantitative EM analysis also provided evidence that many or most of the specific afferent terminals in the damaged VBm were not identical with but were substitutes for the original "vibrissal" specific afferents. Forty percent of all "specific" afferents were shown to be modified corticothalamic terminals. The modification and the resemblence of some cortical endings to specific afferents demonstrated the morphogenetic plasticity of synaptogenesis in these terminals during development as well as the importance and inductive potential of the postsynaptic target in the differentiation of presynaptic axon terminals.

Focal perivascular alterations of white matter in herpes simplex encephalitis--a histological and immunocytochemical study.

Several cases of human herpes simplex encephalitis treated with Vidarabin have been investigated with the histological and immunocytochemical techniques. Cases with a subacute evolution revealed areas of focal perivascular myelin destruction in the white matter. The distribution of herpes simplex antigen did not show any preferential localization of the virus in perivascular oligoglial cells. In contrast, a spatial and temporal relationship has been found between the appearance of immunoglobulin-bearing cells around the vessels and that of areas of focal perivascular myelin damage. Therefore, it is postulated that the areas of focal destruction of myelin are not related to the cytotoxic effect of the virus but are rather dependent on the immune response of the host.

[Organization of the nervous system after coagulation of the follicles of mystacial vibrissae in the newborn mouse: an example of neuronal plasticity]. / L'organisation du système nerveux après coagulation des follicules des vibrisses mystaciales chez la souris nouveau-née: un exemple de plasticité neuronale.

In the mouse the vibrissae and the common fur of the head are a good model of the so called neural plasticity. The characteristics of this model are: the pattern of implantation of the vibrissae at the periphery and that of the arrangement of barrels in the contralateral cortical projection area of vibrissae as well as that of the "barreloïds" in the subcortical vibrissal relays (somato sensory thalamus and trigeminal nuclear complex) are homeomorphic with one another. Each barrel and "barreloïd" receives projections from one vibrissa. Moreover at the level of the cortex these projections are also in register with projections from ipsilateral vibrissae. Head fur hairs project to well defined but entirely distinct areas. Destruction of vibrissae follicles at birth beside preventing barrel and barreloïd formation in the CNS, leads to several morphological changes: degeneration of the primary sensory neurons innervating vibrissae in the trigeminal ganglion, thus degeneration of their central axons and the corresponding terminals in the trigeminal vibrissal relays changes in the distribution of the activity of succinate dehydrogenase in the IVth layer of the cortical vibrissal area and in the corresponding subcortical relays, from the normal discrete (barrel hollow) pattern--corresponding to the clustered vibrissal afferents--to a continuous band, keeping a normal level of activity, excepted in the trigeminal vibrissal relays and a remarkable preservation of cortical thickness but a notable atrophy in the trigeminal vibrissal projection areas. Beside upsetting the anatomy vibrissae follicle destruction causes marked functional changes an outstanding take-over of the deafferented cortical vibrissal area (still identifiable from projections of vibrissae ipsilateral to it) by the head fur hairs this take over exist also in the subcortical vibrissal relays a change in the thalamo-cortical connections. Modifications in the organization of connections are initiated by the loss of the primary sensory neurons innervating vibrissae, in the trigeminal ganglion and results only from early lesions. In mice lesioned when adults the loss of primary sensory neurons is less important and functional take over by the common fur is not observed.

A morphometric study of mouse trigeminal roots after unilateral destruction of vibrissae follicles at birth.

Trigeminal sensory roots were studied in neonatal mice. On the deafferented side, the surface area of the cross-section through the sensory root is diminished by 31% and the number of myelinated fibers is reduced by 21%, but the proportion between myelinated and unmyelinated fibers remains unchanged. The distribution of axonal diameters, analysed in 7 dorso-ventral scanning bands through the sensory roots, indicates a loss or eventually an atrophy of large myelinated axons in the medial two thirds of the sensory root. In both control and deafferented sides the diameter of the myelinated fiber (outside the myelin sheath) is proportional to the axon diameter (inside the myelin sheath). Our results confirm the loss of most of the neurons innervating vibrissae and the lack of regeneration or sprouting in the deafferented root in the newborn mouse.

[Histological changes in the somatosensory thalamus after unilateral coagulation of vibrissa follicles of the muzzle of the newborn mouse]. / Modifications histologiques dans le thalamus somatosensoriel après coagulation unilatérale des follicules des vibrisses du museau chez la Souris nouveaunée.

Light microscopic study of the thalamic ventro-basal complex (VB), after unilateral coagulation of vibrissae follicles in newborn mouse, revealed an excess of neuronal perikarya on the controlateral "deafferented" side as compared to the normal side. The higher density of nerve cells was confined to the vibrissal relay in the medial part of VB nucleus (VBm), whereas the cell number in the non vibrissal-lateral part of this nucleus (VB1) remained on the control level. Electron microscopic investigation of the thalamic vibrissal relay has shown signs of a modified synaptogenesis on the "deafferented" side: (a) the number of specific afferents has diminished and in contrast to the normal side, most of the specific sensory terminals contain small spheroid synaptic vesicles and (b) the number of axon terminals with ovoid pleomorphic vesicles has been doubled.

Histochemical changes of the developing human cerebral neocortex. Studies on two enzymes of energy metabolism in three cortical areas.

Developmental changes of two mitochondrial enzymes, succinate- and menadione-dependent alpha-glycerophosphate dehydrogenases (SDH, M alpha GPDH) have been studied by histochemical techniques in early autopsy material in the following areas of human neocortex: area 4 (motor cortex), area 17 (visual cortex) and area 40 (associative cortex). Each area studied revealed a special enzymatic architecture. SDH - a marker of oxidative activity - develops for several years after birth and in some areas (4 and 40) reaches its adult pattern only after 11 years. Considerable modifications occur during development in the relative activities of the different cortical layers. SDH activity initially appears in layers which send projections to subcortical relays or receive specific thalamocortical afferents. Layers which receive cortico-cortical projections mature later on. In contrast to SDH activity that of M alpha GPDH is elevated already before birth, increases until one month of age and decreases afterwards. This enzyme present in neuroblasts disappears from mature cortical neurons, except from those of the VIth layer. The distribution of this enzyme in cortical layers of the child is opposite to that of SDH. The developmental evolution of these two enzymes has been confirmed by biochemical assays.

Reorganization of thalamo-cortical connections in mice dewhiskered since birth.

Localizations of thalamic relay neurons of vibrissae, common fur of the muzzle and head, as well as those projecting to the cortical vibrissal field in mice with vibrissae coagulated at birth, have been studied by retrograde transport of peroxydase. The tracer was injected in the corresponding cortical areas. In coagulated mice, labeled thalamic neurons were situated in the ventro-basal complex anterior to its vibrissa part, corresponding to the area labeled after intracortical injection of the muzzle and head fur area. These results indicate a reorganization in the origin of thalamo-cortical afferents in coagulated mice.

A morphometric study of mouse trigeminal ganglion after unilateral destruction of vibrissae follicles at birth.

A morphometric study of the trigeminal ganglion after unilateral vibrissae follicles' coagulation in newborn mice has shown the following: (a) a 42.8% decrease of the total volume of the ganglion on the deafferented side with reference to the normal side; a 61.5% decrease of the ophthalmic-maxillary part of the ganglion where neurons whose axons innervate vibrissae follicles are located, and only a 24.1% decrease in the common part; (b) a 54.8% decrease of the neuronal cell body volume in the ophthalmic-maxillary part and practically no change in the common part, and (c) a 64.5% decrease of the volume occupied by the nerve fibers in the ophthalmic-maxillary part and only a 28.1% decrease in the common part. A comparison of the section areas in ganglion and of the bulk area of neuronal cell bodies at different levels has also been performed. Counting of the neuronal cell bodies in the ophthalmic-maxillary part of the ganglion indicated a mean neuronal loss of 36.5%. Peripheral reinnervation of the common fur by regenerated axons of neurons which previously innervated vibrissae, although unlikely, cannot be completely excluded.

Neonatal neuronal necrosis: its relationship to the distribution and maturation of oxidative enzymes of newborn cerebral and cerebellar cortex.

Seventeen brains showing neuronal necrosis after severe perinatal 'asphyxia' were re-examined. In particular the distribution of lesions, which generally involved all layers of the cerebral cortex and was only occasionally laminar, was contrasted with the laminar pattern of maturation of oxidative enzymes in comparable regions of normal newborn brains. In the cerebellum, where necrosis of neurones of the vermis was prominent compared with relative sparing of neurones of the lateral hemispheres, a closer correlation with the pattern of maturation of oxidative enzymes was noted. The rarity of laminar necrosis in the cerebral hemispheres suggests that factors other than pure hypoxic-hypoxia are important in most cases in determining the distribution of lesions, whereas in the cerebellum the earlier maturation of neurones and hypoxic-hypoxia per se are important in determining the localization of lesions.

Delay in the maturation of muscle fibers in infants with congenital hypotonia.

Muscle biopsies of hypotonic children have shown delayed maturation of a fetal type of muscle fibers: subsarcolemmal halo devoid of activity for mitochondrial dehydrogenases, type II predominance and in some cases abnormal dispersion of fiber diameter. Fiber subtypes within group II were also abnormal. One case has definite embryonic characteristics with presence of myoblasts. Not a single clinical pattern was present in these patients and a variety of associated disorders were recognized. Some patients had a clinical picture corresponding to congenital benign hypotonia as described by Walton.

[Histochemical, quantitative and ultrastructural maturation of human fetal muscle]. / Etude de la maturation histochimique, quantitative et ultrastructurale du muscle foetal humain

Histochemical studies of muscle specimens from human fetuses showed: (a) a uniform fiber type population having the properties of Type IIC fibres up to 19 weeks of development; (b) a progressive appearance of Type I fibres after that age; (c) a decrease in number of Type IIC fibres during the last 3 months of pregnancy, accompanied by the appearance of Type IIB and Type IIA fibres; (d) the presence after the myotube stage of fibers with a light peripheral halo in sections stained for mitochondrial dehydrogenases. Electron-microscopic examination of the muscle fibres confirmed the existence of a peripheral halo devoid of myofibrils and mitochondria and showed: (a) scarcity of myofibrils in comparison with mature muscle fibres and (b) irregularity in shape of the myofibrils. In addition, quantitative studies demonstrated an important variation of the fibre diameters up to 21 weeks and the increase of the mean diameter after this age. It is suggested that the persistence after birth of some features of immaturity identical to those described in this work may be considered as a pathological finding.

Vascular and neuroglial changes in experimental herpes simplex encephalitis: ultrastructural study.

Generalized vascular changes and diffused proliferation of reactive microglia were observed in an experimental model of HSV encephalitis of mice. The wide spread of these changes contrasted with the localized character of virus replication and the confined areas of damaged nervous tissue. The vascular and microglial changes were precocious in animals inoculated with concentrated virus suspension (10(5.5)LD50) while they appeared late in mice inoculated with diluted virus suspension (100 LD50). After inoculation with U.V. inactivated virus no changes were seen. The results obtained in this study suggest that the vascular and microglial modifications are not related to a direct cytopathic effect of the virus but dependent on the amount of virus present in the central nervous system and linked to the virus DNA.

Vascular and neuroglial changes in experimental herpes simplex encephalitis enzyme histochemical study.

An enzyme histochemical study of experimental herpes simplex encephalitis of the mouse has revealed a decrease in the number of capillaries displaying alkaline phosphatase activity. Glial cells showed increased Inosine 5 diphosphatase and ATPase activity. These enzyme histochemical changes were distributed throughout the nervous parenchyma while the lesions, seen by light microscopy, are localized to well defined areas. Mice inoculated with a pure culture of irradiated HSV failed to show the above mentioned modifications.