Evidence for an inhibitory effect of protein kinase C on G-protein-mediated repetitive calcium transients in hamster eggs.
Hamster eggs undergo repetitive increases in cytoplasmic free calcium concentration ([Ca2+]i) at fertilization or after injecting guanosine-5'-0-(3-thiotriphosphate) (GTP[S]). We report the effects of protein kinase C (PKC) agonists and antagonists on these repetitive [Ca2+]i transients as measured by their associated membrane potential hyperpolarizing responses (HRs). Iontophoretic injection of GTP[S] into unfertilized eggs caused a series of repetitive HRs that declined in amplitude with time. Continuous injection of inositol 1,4,5-trisphosphate (InsP3) also caused a series of repetitive HRs, but these HRs declined in amplitude less markedly. GTP[S]-induced HRs were inhibited by the PKC agonists phorbol 12-myristate 13-acetate (TPA) (100 nM) and 1,2-dioctanoyl-glycerol (diC8) (250 microM). Conversely the PKC inhibitor sphingosine (10 microM) enhanced the number of large HRs after GTP[S] injection. TPA or sphingosine did not alter InsP3-induced HRs. We suggest that G-protein-mediated InsP3 production causes repetitive [Ca2+]i transients but that GTP[S] injection stimulates a negative feedback loop involving PKC. Adding TPA (100 nM) before insemination caused a reduction in the frequency of HRs at fertilization, but neither TPA nor sphingosine affected the frequency or size of HRs when they were added after the start of fertilization. Fertilizing sperm may stimulate G-protein-mediated InsP3 production in a way that precludes feedback inhibition by PKC.
Swann
K
K
Department of Physiology, Tokyo Women's Medical College, Japan.
Igusa
Y
Y
Miyazaki
S
S
D000818:Animals; D002118:Calcium / Q000037:antagonists & inhibitors / Q000378:metabolism*; D006224:Cricetinae; D004075:Diglycerides / Q000494:pharmacology; D005260:Female; D019204:GTP-Binding Proteins / Q000037:antagonists & inhibitors / Q000502:physiology*; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D006160:Guanosine Triphosphate / Q000494:pharmacology; D015544:Inositol 1,4,5-Trisphosphate / Q000494:pharmacology; D008297:Male; D008564:Membrane Potentials / Q000187:drug effects; D008647:Mesocricetus; D010063:Ovum / Q000187:drug effects / Q000201:enzymology* / Q000378:metabolism; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000502:physiology*; D013110:Sphingosine / Q000494:pharmacology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D013873:Thionucleotides / Q000494:pharmacology
D004075:Diglycerides; D013873:Thionucleotides; C044295:1,2-dioctanoylglycerol; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D015544:Inositol 1,4,5-Trisphosphate; D006160:Guanosine Triphosphate; D011493:Protein Kinase C; D019204:GTP-Binding Proteins; D013110:Sphingosine; D013755:Tetradecanoylphorbol Acetate; D002118:Calcium