Modulation of meiotic arrest in mouse oocytes by guanyl nucleotides and modifiers of G-proteins.
Animal, Cells-Cultured, Cholera-Toxin, Dibutyryl-Cyclic-AMP, Female, Fluorides, FSH, G-Proteins: ph, Graafian-Follicle: ph, Guanine-Nucleotides: ph, Guanosine-Triphosphate: aa, ph, Guanosine-5'-O-(3-Thiotriphosphate): du, Hypoxanthines, Mice, Mice-Inbred-C57BL, Microinjections, Oogenesis: ph, Pertussis-Toxins, SUPPORT-U-S-GOVT-P-H-S, Zona-Pellucida: ph
J Exp Zool 1992 Jul 1;262(4):391-404
Guanyl nucleotide binding-proteins, or G-proteins, are ubiquitous molecules that are involved in cellular signal transduction mechanisms. Because a role has been established for cAMP in meiosis and G-proteins participate in cAMP-generating systems by stimulating or inhibiting adenylate cyclase, the present study was conducted to examine the possible involvement of G-proteins in the resumption of meiotic maturation. Cumulus cell-free mouse oocytes (denuded oocytes) were maintained in meiotic arrest in a transient and dose-dependent manner when microinjected with the nonhydrolyzable GTP analog, GTP gamma S. This effect was specific for GTP gamma S, because GppNHp, GTP, and ATP gamma S were without effect. Three compounds, known to interact with G-proteins, were tested for their ability to modulate meiotic maturation: pertussis toxin, cholera toxin, and aluminum fluoride (AlF4-). Pertussis toxin had little effect on maturation in either cumulus cell-enclosed oocytes or denuded oocytes when meiotic arrest was maintained with dibutyryl cAMP (dbcAMP) or hypoxanthine. Cholera toxin stimulated germinal vesicle breakdown (GVB) in cumulus cell-enclosed oocytes during long-term culture, but its action was inhibitory in denuded oocytes. AlF4- stimulated GVB in both cumulus cell-enclosed oocytes and denuded oocytes when meiotic arrest was maintained with hypoxanthine but was much less effective in dbcAMP-arrested oocytes. In addition, AlF4- abrogated the inhibitory action of cholera toxin in denuded oocytes and also that of follicle-stimulating hormone (FSH) in cumulus cell-enclosed oocytes. Cholera toxin or FSH alone each stimulated the synthesis of cAMP in oocyte-cumulus cell complexes, whereas pertussis toxin or AlF4- alone were without effect. Both cholera toxin and AlF4- augmented the stimulatory action of FSH on cAMP. These data suggest the involvement of guanyl nucleotides and G-proteins in the regulation of GVB, although different G-proteins and mediators may be involved at the oocyte and cumulus cell levels. Cholera toxin most likely acts by ADP ribosylation of the alpha subunit of Gs and increased generation of cAMP, whereas AlF4- appears to act by antagonizing a cAMP-dependent step.
Downs, S M.; Buccione, R; and Eppig, J J., " Modulation of meiotic arrest in mouse oocytes by guanyl nucleotides and modifiers of G-proteins." (1992). Faculty Research 1990 - 1999. 262.