Recent studies have localized gamma-aminobutyric acid (GABA)-containing neurons and identified cells that express subunits of the GABAA receptor in the proliferative zone of the developing cerebral cortex and have demonstrated a role for GABA in cortical neurogenesis. We examined here the interactions between a number of neurotrophic factors, known to be involved in cortical cell proliferation and differentiation, and the GABAergic system (GABA and GABAA receptors) in the regulation of cell production in dissociated cortical cell cultures. We found that basic fibroblast growth factor (bFGF) increased the number of cells labelled for the alpha 1 subunit of the GABAA receptor but not for the alpha 2, alpha 3 or alpha 5 subunits. The alpha 1 subunit was expressed by the majority of proliferating neuroepithelial cells as well as by differentiated neurons. We also found that activation of the GABAA receptor by GABA or muscimol inhibited the proliferative effects of bFGF on cortical progenitors, leading to an increased number of differentiated neurons. These results suggest that bFGF stimulates cell proliferation and GABAA receptor expression in cultured progenitor cells of the developing neocortex, and that GABA regulates cell production by providing a feedback signal that terminates cell division.
The distribution pattern and the morphology of vasoactive intestinal polypeptide (VIP)- and cholecystokinin (CCK)-like-immunoreactive neurons were studied in the brain of the hedgehog and the sheep by means of the peroxidase-antiperoxidase immunocytochemical method. A total of 34 hedgehogs and 26 sheep of both sexes were used. Fourteen hedgehogs and 13 sheep received an intracerebroventricular injection of colchicine that enhanced the immunostaining and revealed "new" immunoreactive cell bodies. VIP-immunoreactive bipolar and multipolar neurons were observed in both species in the cerebral cortex, hippocampal formation, amygdaloid complex, hypothalamus, and central gray substance of the midbrain. CCK-immunoreactive bipolar, bitufted, and multipolar neurons displayed a broader distribution in both mammals than VIP neurons and were found in the cerebral cortex, the hippocampal formation, the amygdaloid complex, the hypothalamus, the mesencephalon, and the pons. In the cortex, in both the hedgehog and the sheep, VIP neurons were located in all layers but were concentrated in layers II and III, with the majority being typical bipolar. CCK neurons were more numerous in the superficial layers (I-III) but were found in the deep layers as well. They were bipolar, bitufted, or multipolar in morphology. From these neurons a small percentage, which were located almost exclusively in layers II and III of the visual cortex, exhibited also VIP immunoreactivity. Perikarya of such double-labeled cells were ovoid or round in shape with one or two main processes emanating from each pole of the cell body and oriented perpendicularly to the pia. The coexistence of the two peptides within individual neurons of the cortex has not been reported in other species and its physiological significance is discussed in relation to the GABAergic neurons of the cortex.
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