Mammalian fertilization encompasses a series of Ca²⁺ oscillations initiated by the sperm factor phospholipase C zeta (PLCζ). Some studies have shown that altering the Ca²⁺ oscillatory regime at fertilization affects preimplantation blastocyst development. However, assisted oocyte activation (AOA) protocols can induce oocyte activation in a manner that diverges profoundly from the physiological Ca²⁺ profiling. In our study, we used the newly developed PLCζ-null sperm to investigate the independent effect of AOA on mouse preimplantation embryogenesis. Based on previous findings, we hypothesized that AOA protocols with Ca²⁺ oscillatory responses might improve blastocyst formation rates and differing Ca²⁺ profiles might alter blastocyst transcriptomes. A total of 326 MII B6D2F1-oocytes were used to describe Ca²⁺ profiles and to compare embryonic development and individual blastocyst transcriptomes between four control conditions: C1 (in-vivo fertilization), C2 (ICSI control sperm), C3 (parthenogenesis) and C4 (ICSI-PLCζ-KO sperm) and four AOA groups: AOA1 (human recombinant PLCζ), AOA2 (Sr²⁺), AOA3 (ionomycin) and AOA4 (TPEN). All groups revealed remarkable variations in their Ca²⁺ profiles; however, oocyte activation rates were comparable between the controls (91.1% ± 13.8%) and AOA (86.9% ± 11.1%) groups. AOA methods which enable Ca²⁺ oscillatory responses (AOA1: 41% and AOA2: 75%) or single Ca²⁺ transients (AOA3: 50%) showed no significantly different blastocyst rates compared to ICSI control group (C2: 70%). In contrast, we observed a significant decrease in compaction (53% vs. 83%) and blastocyst rates (41% vs. 70%) in the absence of an initial Ca²⁺ trigger (AOA4) compared with the C2 group. Transcription profiles did not identify significant differences in gene expression levels between the ICSI control group (C2) and the four AOA groups.

中文翻译：

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在 PLC-Zeta 基因敲除小鼠模型中，根据不同的辅助卵母细胞激活方案对植入前发育进行比较研究。

哺乳动物受精包括由精子因子磷脂酶 C zeta (PLCζ) 引发的一系列 Ca ²⁺振荡。一些研究表明，改变受精时的 Ca ²⁺振荡机制会影响植入前囊胚的发育。然而，辅助卵母细胞激活 (AOA) 协议可以以与生理 Ca ²⁺分析截然不同的方式诱导卵母细胞激活。在我们的研究中，我们使用新开发的 PLCζ-null 精子来研究 AOA 对小鼠植入前胚胎发生的独立影响。基于先前的发现，我们假设具有 Ca ²⁺振荡反应的AOA 协议可能会提高囊胚形成率和不同的 Ca ²⁺配置文件可能会改变囊胚转录组。总共 326 个 MII B6D2F1 卵母细胞用于描述 Ca ²⁺谱并比较四种对照条件之间的胚胎发育和单个囊胚转录组：C1（体内受精）、C2（ICSI 对照精子）、C3（孤雌生殖）和C4（ICSI-PLCζ-KO精子）和四个AOA组：AOA1（人重组PLCζ）、AOA2（Sr ²⁺）、AOA3（离子霉素）和AOA4（TPEN）。所有组都显示出其 Ca ²⁺谱的显着变化；然而，对照组 (91.1% ± 13.8%) 和 AOA (86.9% ± 11.1%) 组之间的卵母细胞活化率相当。使 Ca ²⁺振荡响应（AOA1：41% 和 AOA2：75%）或单一 Ca 的AOA 方法与 ICSI 对照组（C2：70%）相比，2 ⁺瞬变（AOA3：50%）显示没有显着差异的囊胚率。相比之下，我们观察到在没有初始 Ca ²⁺触发 (AOA4)的情况下，与 C2 组相比，压实率（53% 对 83%）和囊胚率（41% 对 70%）显着降低。转录谱没有发现 ICSI 对照组 (C2) 和四个 AOA 组之间基因表达水平的显着差异。