Species-Specific Differences in the Activity and Nuclear Localization of Murine and Bovine Phospholipase C Zeta 1
Species-Specific Differences in the Activity and Nuclear Localization of Murine and Bovine Phospholipase C Zeta 1
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DOI：
10.1095/biolreprod.109.079814
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发表时间：
2010-07-01
影响因子：
3.6
通讯作者：
D'Cruz, Nancy T.
中科院分区：
文献类型：
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作者：
Cooney, Melissa A.;Malcuit, Christopher;D'Cruz, Nancy T.
注射哺乳动物精子提取物或精子特异性磷脂酶C zeta 1（PLCZ 1）的cRNA已被证明会引发游离钙（[Ca2+]（i））浓度的重复振荡，导致迄今为止研究的所有哺乳动物中的卵母细胞活化和胚胎发育。虽然PLCZ 1具有跨物种活性，但也观察到在将PLCZ 1 cRNA注射到不同物种的卵母细胞中后，所产生的[Ca 2 +]（i）振荡的频率和模式可能存在物种特异性差异。因此，我们使用交叉设计策略直接研究鼠和牛PLCZ 1在鼠和牛卵母细胞中的活性。在小鼠卵母细胞中，注射小鼠Plcz1 cRNA诱导[Ca2+]（i）振荡的浓度比牛PLCZ 1低10倍，尽管在牛卵母细胞中，牛PLCZ 1在诱导[Ca2+]（i）振荡方面比小鼠Plcz1更有效。通过PLCZ 1 cRNA对牛卵母细胞中ITPR 1（IP（3）R1）下调的研究也表明，牛PLCZ 1在同源卵母细胞中更有活性。为了确定这些PLCZ是否表现出相似的细胞分布，将Venus标记的PLCZ 1 cRNA注射到卵母细胞中，并使PLCZ 1过表达。牛PLCZ 1未能积累在原核（PN）的牛或小鼠受精卵，尽管拥有一个假定的核定位信号。相反，小鼠PLCZ 1积累在小鼠和牛受精卵的PN中。这些结果表明，小鼠PLCZ 1和牛PLCZ 1具有物种特异性的活性差异，并表明两个物种之间的蛋白质的作用模式的潜在差异。精子PLCZ 1蛋白质含量的物种之间的变化，沿着与卵母细胞的具体差异，在本地化和可用性的PLCZ 1基板，可能进一步有助于优化激活刺激，以提高胚胎发育。
Injection of mammalian sperm extracts or cRNA of the sperm-specific phospholipase C zeta 1 (PLCZ1) has been shown to trigger repetitive oscillations in the concentration of free calcium ([Ca2+](i)), leading to oocyte activation and embryo development in all mammals studied to date. While PLCZ1 has cross-species activity, it has also been observed that species-specific differences may exist in the frequency and pattern of the resulting [Ca2+](i) oscillations following PLCZ1 cRNA injection into oocytes of different species. Accordingly, we used a crossover design strategy to directly investigate the activity of murine and bovine PLCZ1 in both murine and bovine oocytes. In murine oocytes, injection of murine Plcz1 cRNA induced [Ca2+](i) oscillations at 10-fold lower concentrations than bovine PLCZ1, although in bovine oocytes bovine PLCZ1 was more effective than murine Plcz1 at inducing [Ca2+](i) oscillations. Investigation of ITPR1 (IP(3)R1) down-regulation in bovine oocytes by PLCZ1 cRNA also showed that bovine PLCZ1 was more active in homologous oocytes. To determine whether these PLCZs exhibited similar cellular distribution, Venus-tagged PLCZ1 cRNA was injected into oocytes, and PLCZ1 was overexpressed. Bovine PLCZ1 failed to accumulate in the pronucleus (PN) of bovine or murine zygotes, despite possessing a putative nuclear localization signal. Conversely, murine PLCZ1 accumulated in the PN of both murine and bovine zygotes. These results demonstrate that murine PLCZ1 and bovine PLCZ1 possess species-specific differences in activity and suggest potential differences in the mode of action of the protein between the two species. Variation in sperm PLCZ1 protein content among species, along with oocyte-specific differences in the localization and availability of PLCZ1 substrates, may further contribute to optimize the activation stimulus to enhance embryo development.