Jean-François Gadisseux

Effect of ibotenate on brain development: an excitotoxic mouse model of microgyria and posthypoxic-like lesions.

Ibotenate, a glutamatergic agonist, was injected in developing mouse neopallium. When injected at the time of completion of supragranular neuronal migration (P0) ibotenate induces complete neuronal depopulation of layers V-VI and an abnormal sulcation of the overlying supragranular layers. Injected after completion of migration (P5-P10) ibotenate produces severe neuronal loss in layers II, III, IV, V, and VI. After exposure to ibotenate between P0 and P5, surviving neurons have the ability to resume their migration, inducing an abnormal neocortical pattern. Periventricular white matter lesions are observed after ibotenate injection at P2-P10, with a peak of occurrence at P5. Both gray and white matter damage are prevented by DL-2-amino-7-phosphonoheptanoic acid, an N-methyl-D-aspartate receptor antagonist, but not by L (+)-2-amino-3-phosphonopropionic acid, a metabotropic glutamate receptor antagonist. The microtubule-associated type 2 protein, a dendritic marker, is absent in all ibotenate lesions, which reflects the developmental impairment of the dendritic phase. These staged lesions of the gray and white matter disclose a developmental sequence of excitotoxin-affected events starting with the selective and layered sensitivity of postmigratory neocortical neurons and continuing in the white matter with the astroglial maturation and the axonal growth. They faithfully mimic microgyrias, focal cortico-subcortical dysplasias, porencephalic cysts, and white matter damage observed in human perinatal hypoxic/ischemic lesions. This mouse model provides tools for investigating excitotoxic influences on neural development at the various stages and for identifying protective substances against excitotoxicity from hypoxic and from nonhypoxic origins.

The germinative zone produces the most cortical astrocytes after neuronal migration in the developing mammalian brain

The origin of astrocytes of the mouse neocortex during the fetal and early postnatal periods as determined by immunocytological, autoradiographic, electron microscopic and antimitotic methods is described. Most astrocytes destined for the white matter and the infragranular cortical layers are derived from the transformation of radial glial cells between P0 and P10 with an inside-out pattern. This cell metamorphosis is not directly preceded by mitosis and involves the activation of the radial glial lysosomal apparatus. In opposition to recent hypotheses, our findings suggest that most astrocytes destined for the supragranular cortical layers are produced in the germinative zone after the migration of the infragranular neurons and themselves migrate afterwards to the upper cortex between E16 and the first postnatal days. These astrocytes do not display an intermediate stage of the radial glial cell and do not participate in the pattern of appearance of the deeper astrocytes. This second step of astrocytogenesis is a condition for normal cytoarchitectonic development and the maintenance of the supragranular layers, since the deprivation of the astrocytic equipment of the supragranular layers by an antimitotic drug drastically reduces the number of supragranular neurons.

Glial-neuronal Relationship in the Developing Central Nervous-system - a Histochemical Electron Microscope Study of Radial Glial-cell Particulate Glycogen in Normal and Reeler Mice and the Human-fetus

Glial populations in the developing central nervous system can be recognized at the ultrastructural level by its glycogen content stained selectively by reduced osmium and periodic acid-thiocarbohydrazide-silver proteinate. This method permits the description of glial defasciculation during corticogenesis in the normal mouse and demonstrates a disturbance of this process in the reeler mouse. Combined use of autoradiography and glycogen labeling yields insights into the relationships between migrating neurons and radial glial fibers. This method can also be used to recognize and describe glial fibers in the human fetal brain.

Neuron migration within the radial glial fiber system of the developing murine cerebrum an electron microscopic autoradiographic analysis

The present analysis provides direct evidence in the mouse that in the course of course of neocortical histogenesis, contact between migrating neurons and the surfaces of radial glial fibers is both invariant and relatively selective. The analysis characterizes in detail the migratory behavior of the individual migrating cell with respect to the overall radial glial fiber system as this system varies systematically in its structure with ascent through the strata of the cerebral wall. A quantitative study of the relationships between the radial glial fibers confidently identified by their glycogen content and the migrating neurons marked autoradiographically by injection of [3H]thymidine was also performed at the ultrastructural level on tangential sections at different pallial levels in E16 and E17 embryos. The overall set of observations lend support to the hypothesis that radial glial fibers act specifically as guides to neuronal migration and illustrate the nature of the cell-to-cell interaction which serves this cellular process critical to neocortical histogenesis.

Dynamic structure of the radial glial fiber system of the developing murine cerebral wall. An immunocytochemical analysis

Dramatic changes occur in the radial glial fiber system of the murine forebrain in the course of neocortical histogenesis. Initially, prior to substratification of cortical and subcortical anlagen between E13 and E14, the system is uniformly radial in alignment. It appears to achieve maximum density and to be highly uniform in structure throughout its radial span, both in terms of apparent fiber density and the pattern of arrangement of fibers in fascicles. Subsequent to E14, concurrently with rapid growth of the cerebral wall and with the differentiation of the cortical substrata and intermediate zone, the subcortical segment of the system becomes laterally arced while the transcortical span of the system remains radially aligned. Although the spacing between fascicles changes little, there is an apparent general drop in fiber density associated with a progressive reduction in the number of fibers per fascicle. The changes in relative positioning of the fibers and the apparent decline in fiber density are most dramatic within the developing cortical plate and subplate and may be of specific significance for the migration and radial assembly of the neurons in the supragranular neocortical layers.

Topographical and cytological evolution of the glial phase during prenatal development of the human brain: histochemical and electron microscopic study.

An ultrastructural analysis of prenatal gliogenesis and neuronal-glial relationships in the developing fetal brain was carried out using reduced osmium and periodic acid-thiocarbohydrazide-silver proteinate to stain selectively the glycogen content of the glial population. Gliophilic neuronal migration was confirmed in the human fetus, with radial glial fibers (RGF) acting as obligatory corridors for neuronal migration in the prospective neocortex and underlying intermediate zone (IZ). With this method, the entire glial phase was differentiated from neuronal elements; this permitted a description of the evolutionary distribution pattern of RGF: in the cortical plate, glial fascicles fully dissociate by 18 weeks gestation, whereas in the IZ, they remain grouped in fascicles until their transformation into astrocytes. The most conspicuous and constant developmental feature observed in the maturing glial cytoplasm between 21 and 30 weeks gestation was a radical enhancement in the abundance and activity of the lysosomal apparatus and autophagic vacuoles observed in the RGF, a cytological basis for the transformation of radial glial cells into astrocytes. These data have implications for the understanding of the ontogenesis of the neocortical vertical modules in the human brain and for the phylogenetic analysis of the vertical cortical units in terms of comparative mammalian anatomy.

Early neurogenesis and teratogenesis in whole mouse embryo cultures. Histochemical, immunocytological and ultrastructural study of the premigratory neuronal-glial units in normal mouse embryo and in mouse embryos influenced by cocaine and retinoic acid.

Abstract. Yolk sacs of postimplantation mouse embryos were cultured in a mixture of human and rat sera. The central nervous system of these cultured normal embryos was studied from the stage of 5–9 somites (approximately 8.5 postcoital days) to 20–21 somites (approximately 9.5 postcoital days) and compared with in vivo embryos at the same stages. This developmental period covers most of the neural tube closure, the early premigratory differentiation of the neuroectodermal epithelium, and the glial commitment of a population of germinative cells. The neuronal and glial elements of the in vitro cultivated embryos were found to be identical to the corresponding neural tissue in in vivo embryos (light and electron microscopic comparisons); the morphological identity between the in vivo and in vitro embryos was confirmed by morphometry and by stainings revealing the differentiation of the glial elements and precursors. The study of the neuronal-glial units in this material revealed that the fascicular organization of the radial glial cells occurs before the stage of 20 somites. When submitted to a single low dose of retinoic acid at the 7-somite stage, the expression of the epitope recognized by radial cell 2(RC2), a glial marker, is delayed in the in vitro embryos 12–16 hours, but the glycogen and the other glial parameters mature in time. The in vitro embryos exposed to cocaine at the 7-somite stage displayed a prosencephalon remaining deprived of almost all glial cytological features during the entire culture period, although the other developmental parameters evolved normally. This in vitro whole embryo model seems to be a powerful tool for studying early neurogenesis and teratogenesis.

Disorder of cerebellar foliation in Walker's lissencephaly and neu-laxova syndrome.

A diffuse disorder of cerebellar foliation was found in eight infants and one fetus with Walkers lissencephaly. The cerebellar cortex consisted of fused and irregularly distorted folia. In the white matter, trilaminated rings of cortex were concentrically arranged around blood vessels and mesenchymal tissue. The normal relative position of the different classes of cortical nerve cells was preserved. Cells of the external granular layer invaded the meninges and migrated along penetrating blood vessels. We believe that this foliation disorder is caused by a defect in the external basal lamina that allows adjacent folia to be fused and sulci obliterated by intrameningeal ectopias of external granule layer cells. Physical forces applied during development probably contribute to the distortion of the gyral pattern. There was a volumetric reduction of the neocerebellum, which might also be a consequence of the basal lamina defect. The cerebellum of a fetus with the Neu-Laxova syndrome showed the same abnormalities as in Walkers lissencephaly. It is postulated that these two conditions belong to a class of prenatal developmental disorders that involves a defect of the extracellular matrix.

Multiple aggressive vertebral haemangiomas in an adolescent: a case report.

Abstract We describe a neurologically symptom-free adolescent with aggressive vertebral body haemangiomas at two sites.

MR-monitored remitting-relapsing pattern of cortical involvement in Rasmussen syndrome: comparative evaluation of serial MR and PET/SPECT features.

We report the serial MR and PET/SPECT findings in a 2 1/2-year-old boy presenting with Rasmussen syndrome and highlight the close qualitative correlations between the results of the imaging modality and the functional isotopic techniques. The latter demonstrated a wider field of extension of the disease process. Routine MRI demonstrated its ability to detect brain changes matching the more sensitive PET and SPECT data and correlated well with the clinical evolution.