Incompetence of preovulatory mouse oocytes to undergo cortical granule exocytosis following induced calcium oscillations.
Immature oocytes of many species are incompetent to undergo cortical granule (CG) exocytosis upon fertilization. In mouse eggs, CG exocytosis is dependent primarily on an inositol 1,4,5-trisphosphate (IP3)-mediated elevation of intracellular calcium ([Ca2+]i). While deficiencies upstream of [Ca2+]i release are known, this study examined whether downstream deficiencies also contribute to the incompetence of preovulatory mouse oocytes to release CGs. The experimental strategy was to bypass upstream deficiencies by inducing normal, fertilization-like [Ca2+]i oscillations in fully grown, germinal vesicle (GV) stage oocytes and determine if the extent of CG exocytosis was restored to levels observed in mature, metaphase II (MII)-stage eggs. Because IP3 does not stimulate a normal Ca2+ response in GV-stage oocytes, three alternate methods were used to induce oscillations: thimerosal treatment, electroporation, and sperm factor injection. Long-lasting oscillations from thimerosal treatment resulted in 64 and 10% mean CG release at the MII and GV stages, respectively (P < 0.001). Three electrical pulses induced mean [Ca2+]i elevations of approximately 730 and 650 nM in MII- and GV-stage oocytes, respectively, and 31% CG release in MII-stage eggs and 9% in GV-stage oocytes (P < 0.001). Sperm factor microinjection resulted in 86% CG release in MII-stage eggs, while similarly treated GV-stage oocytes exhibited < 1% CG release (P < 0.001). Taken together, these results demonstrate a deficiency downstream of [Ca2+]i release which is developmentally regulated in the 12 h prior to ovulation.
Sackler School of Biomedical Sciences, Tufts University School of Medicine and New England Medical Center, Boston, Massachusetts, 02111, USA.
A L
AL
Abbott
R A
RA
Fissore
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T
Ducibella
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D003594:Cytoplasmic Granules / Q000378:metabolism*; D018274:Electroporation; D005089:Exocytosis / Q000187:drug effects / Q000502:physiology*; D005260:Female; D005456:Fluorescent Dyes / Q000378:metabolism; D005498:Follicular Phase / Q000502:physiology*; D015544:Inositol 1,4,5-Trisphosphate / Q000378:metabolism; D008297:Male; D051379:Mice; D008845:Microinjections; D008856:Microscopy, Fluorescence; D009865:Oocytes / Q000166:cytology* / Q000378:metabolism; D013094:Spermatozoa / Q000378:metabolism; D013849:Thimerosal / Q000494:pharmacology; D015044:Zona Pellucida / Q000187:drug effects
D005456:Fluorescent Dyes; D013849:Thimerosal; D015544:Inositol 1,4,5-Trisphosphate; D002118:Calcium