PLC? can stably regulate Ca2+ fluctuations in early embryo

Phospholipase C zeta (PLC ) is an important inducer of Ca2++ oscillations in mammalian sperm. To explore the influence of PLC on early embryonic Ca2+ fluctuations during sperm-egg binding, this study used PLC from sheep sperm to construct an early embryonic Ca2+ fluctuation model. First, sheep MII oocytes were cultivated and screened using microinjection technology. Then, a pEGFP-N1-PLC plasmid was constructed to activate oocytes in the test group. Ionomycin combined with 6-Dimethylaminopurine (6-DMAP) was used for the control group to explore the effects on early embryonic development and regulation of Ca2+ fluctuations during development. The results demonstrated that both the PLC and ionomycin combined with 6-DMAP activation methods induced sheep oocyte parthenogenetic activation and development in early embryos. In comparisons, the cleavage rate of ionomycin combined with 6-DMAP activation was significantly higher than that of PLC (60.9% +/- 19.4% vs 76.1% +/- 0.7%, respectively; p < 0.001), and the blastocyst rates were 16.2% +/- 0.62% and 21.1% +/- 0.92%, respectively (p < 0.05). Additionally, when comparing the distribution of Ca2+ in early embryos at different stages, Ca2+ in both treatment groups was mainly distributed in the cytoplasm, but the temporal pattern of Ca2+ fluctuations differed. PLC resulted in Ca2+ peaks that appeared at the cleavage and morula stages of early embryos, and Ca2+ returned to normal levels at the morula stage. However, the Ca2+ concentration after ionomycin combined with 6-DMAP activation was always much higher than that with PLC , and its single peak appeared later than in the PLC group. In summary, the PLC gene promoted stable regulatory effects on Ca fluctuations at different stages during early embryonic development.(c) 2022 Elsevier Inc. All rights reserved.