title: 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 internal Galpha(s) after treatment of oocytes with progesterone correlated with the steroid-induced maturation response, and the increase in internal Galpha(s) after hormone treatment was comparable to the decrease in cortical Galpha(s). These results are consistent with a model in which release of Galpha(s) from the plasma membrane is involved in activation of the progesterone-signaling pathway that leads to amphibian oocyte maturation.
authors.affiliation: Department of Biological Sciences, University of Denver, Denver, Colorado 80208, USA. ssadler@du.edu
authors.forename: Susan E
authors.initials: SE
authors.lastname: Sadler
authors.forename: Mallory R
authors.initials: MR
authors.lastname: Archer
authors.forename: Kirsten M
authors.initials: KM
authors.lastname: Spellman
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism; D002469:Cell Separation; D006063:Chorionic Gonadotropin / Q000494:pharmacology; D005260:Female; D019205:GTP-Binding Protein alpha Subunits, Gs / Q000032:analysis / Q000378:metabolism*; D008540:Meiosis / Q000187:drug effects; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D009865:Oocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D006080:Ovarian Follicle / Q000166:cytology; D010766:Phosphorylation / Q000187:drug effects; D011374:Progesterone / Q000494:pharmacology*; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D021381:Protein Transport / Q000187:drug effects; D017231:Proto-Oncogene Proteins c-mos / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects* / Q000502:physiology; D013045:Species Specificity; D013256:Steroids / Q000494:pharmacology*; D029867:Xenopus Proteins / Q000378:metabolism; D014982:Xenopus laevis; D047392:beta-Cyclodextrins / Q000494:pharmacology*
chemical_list: D006063:Chorionic Gonadotropin; D011500:Protein Synthesis Inhibitors; D013256:Steroids; D029867:Xenopus Proteins; D047392:beta-Cyclodextrins; C108732:methyl-beta-cyclodextrin; D011374:Progesterone; C495329:Mos protein, Xenopus; D017231:Proto-Oncogene Proteins c-mos; D020928:Mitogen-Activated Protein Kinases; D019205:GTP-Binding Protein alpha Subunits, Gs