Membrane binding sites and non-genomic effects of estrogen in cultured human preosteoclastic cells

https://doi.org/10.1016/S0960-0760(96)00092-1Get rights and content

Abstract

Besides functional estrogen receptors, the presence of signalling cell surface binding sites for 17β-estradiol (17βE2) has been reported in osteoblast- and osteoclast-like cells, suggesting that 17βE2 may influence bone remodelling by a dual mechanism of action: to affect gene expression mediated by the nuclear activity of the steroid-receptor complex, and to initiate rapid responses triggered by a signal-generating receptor on the cell surface. Recently, we demonstrated that the human preosteoclastic cell line FLG 29.1 bears functional estrogen receptors. In this study we examined FLG 29.1 cells for the presence of cell surface binding sites for 17βE2, and whether 17βE2 could elicit cell signalling. Using a cell-impermeant and fluorescent estrogen conjugate, 17β-estradiol-6-carboxy-methyloxime-bovine serum albumin-fluorescein isothiocyanate, we demonstrated the presence of specific plasma membrane binding sites for 17βE2. Stimulation of FLG 29.1 cells with low (1 nM) and high (1 μM) doses of 17βE2 induced a prompt and significant (P < 0.05) increase of cellular pH, as measured in single cells using an image analysis system. In addition, both cAMP and cGMP were significantly increased by 17βE2 with a dose-dependent response. Finally, a rapid increase of intracellular calcium ion concentration [Ca2+] was also induced by 1 nM 17βE2, as measured in single cells using an image analysis system. Our findings strongly suggest a non-genomic action of 17βE2 on osteoclast precursors.

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