Immunohistochemical study of Pax6 and Reissner fibre expression in the prenatal development of the mouse subcommissural organ

The subcommissural organ (SCO) releases glycoproteins into the ventricular cerebrospinal fluid (CSF), where they form Reissner's fibre (RF) and also secretes a CSF-soluble material different from RF-material. Pax6 is a transcription factor important for the regulation of cell proliferation, migration and differentiation in the developing brain. In the present work, we studied wild-type, heterozygous and homozygous Sey mice to compare the expression of RF-antibody and Pax6 in the SCO and adjacent structures. In wild-type mice between E15 to E18, we observed Pax6 expression in cells surrounding the secretory cells of the SCO, and RF-immunoreactive material only in the SCO ependymal cell layer and its basal process. In the heterozygous mice, the neuroanatomical structure of the SCO was present, but RF-antibody staining and Pax6 expression was scarce or almost undetectable; in the homozygous mice neither SCO nor other epithalamic structures were found. We suggest that Pax6 expression at the periphery of the SCO is essential for the development and activity of the organ (AU)

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Cloning and cortical expression of rat Emx2 and adenovirus-mediated overexpression to assess its regulation of area-specific targeting of thalamocortical axons.

A goal of this study was to use recombinant adenovirus (AdV) to ectopically express Emx2 in the embryonic neocortex as a gain-of-function approach to study its role in the area-specific targeting of thalamocortical axons (TCAs), using the rat as a model. First, we cloned the cDNA for the full-length coding region of rat Emx2, a homologue of Drosophila empty spiracles. We also used this sequence to define the full-length coding region of mouse Emx2 from genomic DNA. Our analysis of Emx2 expression shows that in rat, as reported in mouse, Emx2 is expressed in high caudal to low rostral, and high medial to low lateral, gradients across the cortex throughout cortical neurogenesis, and expression is primarily restricted to progenitors in the neuroepithelium. We also carried out an analysis of the distribution of cells infected with a replication defective recombinant type 5 adenovirus (AdV) containing a CAG/LacZ expression construct, following an injection into the lateral ventricle of the cerebral hemisphere at different stages of embryonic cortical neurogenesis. AdV-infected cells are broadly distributed tangentially, but their laminar distribution is differentially restricted and reflects the temporal sequence of generation of cortical neurons. This finding indicates that the AdV predominantly infects progenitors in the ventricular zone, which leads to a preferential labeling of their immediate progeny, and infects cells that have recently become postmitotic and have yet to move far from the ventricular surface. We then show that AdV-mediated ectopic Emx2 expression results in aberrant intracortical pathfinding and areal targeting of TCAs from the dorsal lateral geniculate nucleus. These findings indicate that EMX2 imparts positional information normally associated with caudal cortical areas, such as the primary visual area, that influences the area-specific targeting of TCAs. These results are consistent with a role for EMX2 in areal specification of the neocortex as suggested by recent analyses of Emx2 null mutants.

Cell-cycle kinetics of neocortical precursors are influenced by embryonic thalamic axons.

Thalamic afferents are known to exert a control over the differentiation of cortical areas at late stages of development. Here, we show that thalamic afferents also influence early stages of corticogenesis at the level of the ventricular zone. Using an in vitro approach, we show that embryonic day 14 mouse thalamic axons release a diffusable factor that promotes the proliferation of cortical precursors over a restricted developmental window. The thalamic mitogenic effect on cortical precursors (1) shortens the total cell-cycle duration via a reduction of the G(1) phase; (2) facilitates the G(1)/S transition leading to an increase in proliferative divisions; (3) is significantly reduced by antibodies directed against bFGF; and (4) influences the proliferation of both glial and neuronal precursors and does not preclude the action of signals that induce differentiation in these two lineages. We have related these in vitro findings to the in vivo condition: the organotypic culture of cortical explants in which anatomical thalamocortical innervation is preserved shows significantly increased proliferation rates compared with cortical explants devoid of subcortical afferents. These results are in line with a number of studies at subcortical levels showing the control of neurogenesis via afferent fibers in both vertebrates and invertebrates. Specifically, they indicate the mechanisms whereby embryonic thalamic afferents contribute to the known early regionalization of the ventricular zone, which plays a major role in the specification of neocortical areas.

Normal fetal brain development: MR imaging with a half-Fourier rapid acquisition with relaxation enhancement sequence.

PURPOSE: To analyze normal maturation of the fetal brain with half-Fourier rapid acquisition with relaxation enhancement (RARE) magnetic resonance (MR) imaging. MATERIALS AND METHODS: The normal brains of 25 fetuses of 12-38 weeks gestational age were examined in utero with half-Fourier RARE imaging. Gyrus maturation, gray and white matter differentiation, ventricle-to-brain diameter ratio, and subarachnoid space size were evaluated with respect to gestational age. RESULTS: At 12-23 weeks, the brain had a smooth surface, and two or three layers were differentiated in the cerebral cortex. At 24-26 weeks, only a few shallow grooves were seen in the central sulcus, and three layers, including the immature cortex, intermediate zone, and germinal matrix, were differentiated in all fetuses. At 27-29 weeks, sulcus formation was observed in various regions of the brain parenchyma, and the germinal matrix became invisible. Sulcation was seen in the whole cerebral cortex from 30 weeks on. However, the cortex did not undergo infolding, and opercular formation was not seen before 33 weeks. At 23 weeks and earlier, the cerebral ventricles were large; thereafter, they gradually became smaller. The subarachnoid space overlying the cortical convexities was slightly dilated at all gestational ages, most markedly at 21-26 weeks. CONCLUSION: Changes in brain maturation proceed through stages in an orderly and predictable fashion and can be evaluated reliably with half-Fourier RARE MR imaging.

Third ventricle: size and appearance in normal fetuses through gestation.

PURPOSE: To define the size and appearance of the normal fetal third ventricle. MATERIALS AND METHODS: The third ventricle was prospectively assessed in 441 consecutive normal second- and third-trimester fetuses. The fetuses were divided into six gestational age ranges. Data regarding the size and configuration of the third ventricle were analyzed for each group. RESULTS: The third ventricle was seen in 435 of 440 (98.9%) fetuses. It appeared as a single echogenic line between the thalami in 171 (38.9%) fetuses, as parallel echogenic lines outlining a fluid-filled lumen in 243 (55.2%) fetuses, and as divergent lines delineating a V-shaped fluid-filled structure in 21 (4.8%) fetuses. The single-line configuration was most common early in the second trimester. Later in pregnancy, the ventricle walls could be discerned as separate parallel or divergent lines outlining a fluid-filled lumen. The average width of the ventricle was relatively constant at approximately 1 mm from 12 to 28 weeks. After this time, it enlarged, reaching a maximum 1.9 mm. CONCLUSION: The third ventricle can be imaged in most second- and third-trimester fetuses. Its size and configuration evolve through the second and third trimesters. This evolution must be considered in the evaluation of normality. At any gestational age, a third ventricle greater than 3.5 mm in width should be viewed with concern for abnormality.

[Immunocytochemical and autoradiographic research on the growth of serotoninergic fibers to the cerebral ventricles in the perinatal period in rats]. / Immunotsitokhimicheskoe i avtoradiograficheskoe issledovaniia prorastaniia serotoninergicheskikh volokon k zheludochkam mozga v perinatal'nom periode u krys.

Anatomical relationships between serotoninergic (5-HT) fibers and cerebral ventricles were studied in rats from the 16th fetal day until the 9th postnatal day with immunocytochemistry and radioautography. In the latter case, 5-HT neuronal elements were detected according to their specific uptake of intraventricularly injected 3H-5-HT. On the 16th fetal day, occasional 5-HT fibers first spread from the main place of their origin in the raphe nuclei to the dorsocaudal portion of the 3rd ventricle and aqueduct. Two days later, a more extensive network of 5-HT fibers appeared around the dorsal portion of the 3rd ventricle, whereas fibers only rarely penetrated fibers became noticeable in the lateral and 3rd ventricles. The functional significance of hypothalamic and ventricular 5-HT is discussed from the standpoint of its being either a modulator of growth and differentiation of the developing brain, or a factor involved in some specific neuroendocrine functions.

[Participation of the tanycytes in the mechanisms of sex differentiation of the brain]. / Uchastie tanitsitov v mekhanizmakh polovoi differentsirovki mozga.

Enzymo-histochemical methods were applied to the study of the activity of a number of dehydrogenases of the specialized ependyma of the inferior-lateral walls and floor of the 3rd ventricle in rats of both sexes during the "critical" period of sexual differentiation of the hypothalamic region of the brain (the first week after birth). At the level of the arcuate nucleus (alpha2-tanycytes) and the medial portion of the medial eminence (beta2-tanycytes) the ependyma was characterized by similar metabolic activity indices in ratlings of both sexes at the decisive periods of the "critical" period (the 3rd, 5th, 7th days after birth). The metabolic activity was reduced on the 5th day in the cells of animals of both sexes. At the critical period the ependyma of the lateral portions of the median eminence (beta1-tanycytes) displayed marked sexual differences in the activity of the enzymes under study. On the 5th day beta1-tanycytes of male rats showed a reduction and of female rats - an elevation of the metabolic activity. The results obtained permit to suppose that beta1-tanycytes participated in the mechanisms of sexual differentiation of the brain.