Insulin influences astroglial morphology and glial fibrillary acidic protein (GFAP) expression in organotypic cultures.
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abstract
Variations in the levels and timing of exposure to insulin-related peptides influence the phenotypic appearance of astroglia present in organotypic cultures of the E17 mouse cerebellum as well as the expression of glial fibrillary acidic protein (GFAP) mRNA and its encoded protein. The morphology of GFAP-immunoreactive cells was influenced by the levels of insulin added in an age-specific manner. Fetal radial glia were selectively and significantly (P less than 0.001) increased by high (10 micrograms/ml) insulin levels, comprising the majority of the GFAP-positive cells seen. In contrast, there was an almost complete reversal of this pattern elicited by low (10 pg/ml) insulin levels, where GFAP-positive cells appeared undifferentiated and epithelioid (P less than 0.001). In newborn cultures, on the other hand, the morphological responses to both high and low levels of insulin were considerably attenuated and involved radial glia primarily, whose numbers were significantly increased by the high insulin levels. Exposure to high levels of insulin was accompanied by an increase in GFAP mRNA expression, as determined by non-isotopic (biotin) in situ hybridization histochemistry, and intense GFAP immunoreactivity, while low insulin levels elicited minimal expression of both message and protein product. In view of the critical interdependence of developing neurons and radial glia with respect to neuronal migration and the differentiation of neurons and astroglia, the responses observed suggest developmentally regulated mechanisms by which insulin-related peptides themselves may influence directly and indirectly both neuronal and astroglial differentiation.
