Activation of the progesterone-signaling pathway by methyl-beta-cyclodextrin or steroid in Xenopus laevis oocytes involves release of 45-kDa Galphas.
Abstract
Treatment of Xenopus laevis oocytes with cholesterol-depleting methyl-beta-cyclodextrin (MebetaCD) stimulates phosphorylation of mitogen-activated protein kinase (MAPK) and oocyte maturation, as reported previously [Sadler, S.E., Jacobs, N.D., 2004. Stimulation of Xenopus laevis oocyte maturation by methyl-beta-cyclodextrin. Biol. Reprod. 70, 1685-1692.]. Here we report that treatment of oocytes with MebetaCD increased levels of immunodetectable 39-kDa mos protein. The protein synthesis inhibitor, cycloheximide, blocked the appearance of Mos, blocked MebetaCD-stimulated phosphorylation of MAPK, and inhibited MebetaCD-induced oocyte maturation. These observations suggest that MebetaCD activates the progesterone-signaling pathway. Chemical inhibition of steroid synthesis and mechanical removal of follicle cells were used to verify that MebetaCD acts at the level of the oocyte and does not require production of steroid by surrounding follicle cells. Cortical Galpha(s) is contained in low-density membrane; and treatment of oocytes with progesterone or MebetaCD reduced immunodetectable levels of Galpha(s) protein in cortices and increased internal levels of 45-kDa Galpha(s) in cortical-free extracts. Dose-dependent increases in inte...Continue Reading
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