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• W2595802676 abstract "During fertilization, sperm PLC-zeta induces hydrolysis of phosphoinositide in diacylglycerol and inositol 1,4,5-trisphosphate (IP3). IP3 triggers Ca2+ release by binding to IP3 receptor (IP3R-1), starting a series of intracellular calcium ([Ca2+]i) oscillations that is responsible for oocyte activation and initiation of embryonic development. These oscillations start at metaphase II (MII) and finish as the zygotes progress into interphase and initiate pronuclear formation when IP3R-1 undergoes degradation. However, the role of IP3-R1 in embryo development is not known. Since as the function of IP3-R1 in embryo development is unknown, we are interested in the role of IP3R-1 after zygote stage. To address these issues, the aim of the present study was to evaluate the zygote IP3R-1 down-regulation on subsequent embryo development comparing mice zygote microinjected with adenophostin A (20µM) to control mice zygote microinjected with buffer. The western blot analysis showed that adenophostin microinjection efficiently induced IP3R-1 down-regulation in zygotes when compared to control zygotes (tree replicates per group with 25 zygotes each). The adenophostin is known to induce Ca2+ release during IP3R-1 degradation. In this manner, we evaluated the adenophostin ability to induce Ca2+ release during zygote IP3R-1 degradation. Briefly, MII oocytes and zygotes were microinjected with adenophostin and Ca2+ oscillations measured for 1 hour. As expected, adenophostin microinjection initiated [Ca2+]i oscillations at the MII stage but [Ca2+]i oscillations were absent in zygote microinjected with adenophostin (approximately 10 oocytes and 10 zygotes were evaluated). Once we have verified that adenophostin degraded IP3R-1 and this degradation did not initiate [Ca2+]i oscillations, zygotes microinjected with adenophostin or buffer were cultured for 5 days to evaluated zygote IP3R-1 down-regulation effect on embryo development. The embryo development rate from 82 zygotes microinjected with adenophostin and 74 with buffer were analyzed by Chi-square. Adenophostin microinjection resulted in 2-cell stage embryos rate that were similar to control zygotes (96.2 vs. 96.4% respectively). However, 4 cells stage embryos were significantly lower in zygotes microinjected with adenophostin compared to control group (42.2 vs. 79.7%, respectively; P<0.01). Despite the reduction in number of 4 cells stage embryos from zygotes microinjected with adenophostin, it appears that these embryos recovered their development capacity once they reached morulae stage in a similar way to control group (66.2 vs. 76%, respectively). Likewise, when the number of embryos that developed to the blastocyst stage at day 4 (56.5 vs. 72.5%) and day 5 (64.2 vs. 77.9%) was evaluated, no differences were observed between groups. After embryo development evaluation, the cell number of these embryos were measured. Approximately 15 embryos at day 5 from each group were stained with hoechst and the cell number evaluated. No difference was observed between the two groups (57 vs. 69.5%) on embryo quality. The results showed that IP3R-1 down-regulation in zygotes affects the embryo development without influence of [Ca2+]i oscillations. However, these embryos recovered their development capacity throughout in vitro culture. It still unclear if the importance of IP3R-1 is only at 4 cells stage or if these embryos reorganize or recovery the number of IP3R-1 mass during their development. (poster)" @default.
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• W2595802676 date "2009-07-01" @default.
• W2595802676 modified "2023-10-16" @default.
• W2595802676 title "Mouse Zygote IP3 Receptor Down-Regulation Affects Subsequent Embryo Development." @default.
• W2595802676 doi "https://doi.org/10.1093/biolreprod/81.s1.243" @default.
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