Competence to undergo normal, fertilization-induced cortical activation develops after metaphase I of meiosis in mouse oocytes.
The purpose of this study was to determine at what stage of meiotic maturation mouse oocytes develop the ability to undergo sperm-induced activation, assayed by cortical granule (CG) release. Germinal vesicle breakdown (GVBD), prometaphase I (proMI), metaphase I (MI), and metaphase II (MII, the normal stage at fertilization) stages were evaluated by quantitative image analysis. At 2 hr, fertilized MII eggs underwent a mean CG loss of 69%; loss was global--in the entire cortex occupied by CGs. In contrast, fertilized GVBD and proMI oocytes had no significant CG loss in the cortex after 2 hr. After 4 hr, 63, 67, and 88% of fertilized GVBD, proMI, and MI oocytes, respectively, had localized CG release only in the vicinity of the fertilizing sperm. At 4 hr, GVBD oocytes had small sperm-associated, CG-depleted domains, approximately 100 microns 2 (< 1% of the oocyte cortex), whereas those for proMI oocytes were approximately 2500 microns 2. The CG density was reduced by 90% in these domains, whereas the remaining cortex showed no evidence of CG loss. Both the extent and time course of CG loss were altered in oocytes fertilized before MII and were not significantly affected with 25 microM thimerosal, which sensitizes egg calcium stores. Unlike the ability to undergo CG release, the ability to initiate sperm chromatin decondensation was not stage specific and the extent of decondensation was cell cycle related. Competence to undergo CG loss develops in two phases: the ability to undergo localized CG release increases between the GV and MI stages, whereas the mechanism of propagating a normal wave of global CG loss from the site of sperm entry develops between MI and MII.
Ducibella
T
T
Department of Obstetrics and Gynecology (Division of Reproductive Endocrinology), Tufts University School of Medicine, Boston, Massachusetts 02111.
Buetow
J
J
D000818:Animals; D015550:Cell Degranulation; D002478:Cells, Cultured; D002843:Chromatin; D005260:Female; D005306:Fertilization; D008297:Male; D008540:Meiosis; D008677:Metaphase; D051379:Mice; D009865:Oocytes / Q000166:cytology* / Q000187:drug effects; D013094:Spermatozoa / Q000166:cytology; D013849:Thimerosal / Q000494:pharmacology
D002843:Chromatin; D013849:Thimerosal