Role of G proteins in mouse egg activation: stimulatory effects of acetylcholine on the ZP2 to ZP2f conversion and pronuclear formation in eggs expressing a functional m1 muscarinic receptor.
Sperm-mediated egg activation may be analogous to ligand-mediated signal transduction through G protein-coupled receptors. We investigated this possibility in the mouse egg by microinjecting mouse oocytes with an m1 muscarinic receptor mRNA. Following oocyte maturation in vitro, the metaphase II-arrested eggs were treated with acetylcholine and its effect was examined on zona pellucida modifications and pronuclear formation, which are end points of early and late egg activation, respectively. Treatment of these eggs with acetylcholine reveals that both the ZP2 to ZP2f conversion and pronuclear formation occur. Atropine and microinjected GDP beta S block the acetylcholine-induced ZP2 conversion, suggesting that the acetylcholine effects are mediated via a functional G protein-coupled m1 receptor. The acetylcholine-induced ZP2 conversion, however, is not inhibited by pertussis toxin under conditions in which greater than 90% of the endogenous Gi is inactivated by ADP ribosylation. The presence of a pertussis toxin-insensitive G protein, Gq, is detected by immunoblotting; this G protein could be a candidate to mediate the pertussis toxin-insensitive effects of acetylcholine. Results of these experiments are consistent with the hypothesis that receptor-mediated G protein activation may play a role in egg activation.
Williams
C J
CJ
Department of Obstetrics and Gynecology, University of Pennsylvania, Philadelphia 19104.
Schultz
R M
RM
Kopf
G S
GS
D000109:Acetylcholine / Q000494:pharmacology*; D000818:Animals; D001285:Atropine / Q000494:pharmacology*; D004527:Egg Proteins; D005260:Female; D019204:GTP-Binding Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D006023:Glycoproteins / Q000378:metabolism*; D006153:Guanosine Diphosphate / Q000031:analogs & derivatives / Q000494:pharmacology; D008297:Male; D008562:Membrane Glycoproteins; D051379:Mice; D008845:Microinjections; D008954:Models, Biological; D009865:Oocytes / Q000187:drug effects* / Q000378:metabolism; D011956:Receptors, Cell Surface; D011976:Receptors, Muscarinic / Q000032:analysis* / Q000235:genetics; D013084:Sperm-Ovum Interactions / Q000187:drug effects; D013873:Thionucleotides / Q000494:pharmacology; D015044:Zona Pellucida / Q000187:drug effects* / Q000378:metabolism; D000072216:Zona Pellucida Glycoproteins
D004527:Egg Proteins; D006023:Glycoproteins; D008562:Membrane Glycoproteins; D011956:Receptors, Cell Surface; D011976:Receptors, Muscarinic; D013873:Thionucleotides; D000072216:Zona Pellucida Glycoproteins; C000606520:Zp2 protein, mouse; D006153:Guanosine Diphosphate; C023427:guanosine 5'-O-(2-thiodiphosphate); D001285:Atropine; D019204:GTP-Binding Proteins; D000109:Acetylcholine