Intracellular calcium responses in bovine oocytes induced by spermatozoa and by reagents.
The objectives of the present study were to clarify and compare the characteristics of the transient rises in intracellular calcium concentrations ([Ca2+]i) induced either by spermatozoa or by stimulation with artificial activators in bovine oocytes. These transient rises in [Ca2+]i in oocytes matured in vitro were recorded with Ca2+ imaging using the Ca2+ indicator fura-2. During fertilization, a series of transient rises in [Ca2+]i was observed. The first Ca2+ response peaked at a concentration of 521 +/- 39 nM (n = 20) and lasted for 4 min, while the subsequent Ca2+ responses were significantly smaller and shorter, with a peak of 368 +/- 13 nM (n = 23) and a duration of 2 min. Injection of inositol 1,4,5- triphosphate (InsP3) into unfertilized oocytes caused a transient rise in [Ca2+]i in a dose-dependent manner. The maximum response was induced by 20 nA x 1 sec injection of InsP3. Thimerosal, a sulfhydryl reagent, induced the repetitive transient rises in [Ca2+]i. The peak and the duration of the rises in [Ca2+]i induced by InsP3 or thimerosal were smaller and shorter, respectively, than those of the first rise induced by spermatozoa. Ethanol and Ca2+ ionophore IA23187, which are general parthenogenetic activators of unfertilized oocytes, each induced a single transient rise in [Ca2+]i. The duration of the rise in [Ca2+]i by ethanol or Ca2+ ionophore was significantly longer than that by spermatozoa at fertilization, although the peaks were smaller. These results clarified the characteristics of the rises in [Ca2+]i induced by spermatozoa and by several artificial reagents, and showed that the first rise in [Ca2+]i induced by spermatozoa had a higher peak [Ca2+]i and a longer duration compared with each the subsequent rises in [Ca2+]i and the rises in [Ca2+]i induced by artificial reagents. These indicate that a mode like as the first rise in [Ca2+]i induced by spermatozoa is an effective trigger for artificial activation of oocytes.
Nakada
K
K
Department of Veterinary Obstetrics and Gynecology, Rakuno Gakuen University, Hokkaido, Japan.
Mizuno
J
J
D000818:Animals; D000001:Calcimycin / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D002417:Cattle / Q000378:metabolism*; D000431:Ethanol / Q000494:pharmacology; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary; D007202:Indicators and Reagents / Q000494:pharmacology*; D007476:Ionophores / Q000494:pharmacology; D008297:Male; D009865:Oocytes / Q000187:drug effects / Q000378:metabolism*; D010312:Parthenogenesis; D013084:Sperm-Ovum Interactions; D013849:Thimerosal / Q000494:pharmacology
D007202:Indicators and Reagents; D007476:Ionophores; D013849:Thimerosal; D000001:Calcimycin; D000431:Ethanol; D002118:Calcium