We investigated the effect of the female hormone 17β-estradiol (E2) and the hormone mimic bisphenol A (BPA) on the proliferation and differentiation of rat neural stem/progenitors cells (NS/ PCs) cultured from the telencephalon of embryonic day-15 rats. Basic fibroblast growth factor (FGF-2) is a potent mitogen of early generated NS/PCs, and is used for the proliferation of NS/ PCs in vitro. Administration of E2 or BPA alone to the NS/PCs stimulated their proliferation in the absence but not in the presence of FGF-2. E2-or BPA-treatment increased the ratio of the oligodendrocytes generated from the NS/PCs to total cells; however, this ratio did not change when the cells were stimulated with platelet-derived growth factor (PDGF), a mitogen for oligodendrocyte precursors, or with neurotrophin-3, an oligogenic factor for glial progenitor cells. These results suggest that estrogens would influence the fate of NS/PCs when the cells are poorly supplied with mitogens or differentiation factors during the early stages of neurogenesis.

Our previous study indicated that both 17β-estradiol (E2), known to be an endogenous estrogen, and bisphenol A (BPA), known to be a xenoestrogen, could positively influence the proliferation or differentiation of neural stem/progenitor cells (NS/PCs). The aim of the present study was to identify the signal transduction pathways for estrogenic activities promoting proliferation and differentiation of NS/PCs via well known nuclear estrogen receptors (ERs) or putative membrane-associated ERs. NS/PCs were cultured from the telencephalon of 15-day-old rat embryos. In order to confirm the involvement of nuclear ERs for estrogenic activities, their specific antagonist, ICI-182,780, was used. The presence of putative membrane-associated ER was functionally examined as to whether E2 can activate rapid intracellular signaling mechanism. In order to confirm the involvement of membrane-associated ERs for estrogenic activities, a cell-impermeable E2, bovine serum albumin-conjugated E2 (E2-BSA) was used. We showed that E2 could rapidly activate extracellular signal-regulated kinases 1/2 (ERK 1/2), which was not inhibited by ICI-182,780. ICI-182,780 abrogated the stimulatory effect of these estrogens (E2 and BPA) on the proliferation of NS/PCs, but not their effect on the differentiation of the NS/PCs into oligodendroglia. Furthermore, E2-BSA mimicked the activity of differentiation from NS/PCs into oligodendroglia, but not the activity of proliferation. Our study suggests that (1) the estrogen induced proliferation of NS/PCs is mediated via nuclear ERs; (2) the oligodendroglial generation from NS/PCs is likely to be stimulated via putative membrane-associated ERs.

Protein-protein interactions among 960 Pyrococcus soluble proteins have been analysed by mammalian two-hybrid analysis and thirteen interactions between annotated and unannotated proteins detected.