Spindlin I is essential for metaphase II stage maintenance and chromosomal stability in porcine oocytes
Spindlin I is essential for metaphase II stage maintenance and chromosomal stability in porcine oocytes
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DOI：
10.1093/molehr/gax005
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发表时间：
2017-03-01
影响因子：
4
通讯作者：
Cui, Xiang-Shun
中科院分区：
文献类型：
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作者：
Choi, Jeong-Woo;Zhao, Ming-Hui;Cui, Xiang-Shun
研究问题：Spindlin I（Spin I）在猪中期II期卵母细胞中的功能是什么？简要答复：Spin I的耗竭诱导自发的卵母细胞活化，Spin I的过表达通过诱导猪卵母细胞DNA损伤导致多核形成。在小鼠卵母细胞中，Spin 1在配子发生过程中特异性表达，并且对于减数分裂的恢复至关重要。在体细胞中，SpinI促进癌细胞增殖并激活WNT/T细胞因子signaling.Study设计大小，持续时间：在猪MII期卵母细胞中敲除（KD）或过表达SpinI后，在额外培养24小时后检查MII维持。研究单性生殖细胞培养到4个细胞（72 h）或囊胚（7 d）stage.PARTICIPANTS/MATERIALS，SETTING，METHODS：通过显微注射猪SpinI靶向siRNA，在猪卵母细胞和胚胎中敲低SpinI。对于Spin 1过表达，产生猪Spin l-eGFP cRNA。此外，对于拯救实验，将编码siRNA抗性小鼠SpinI的cRNA添加到猪SpinI靶向siRNA中。对于过表达和救援实验，显微注射和文化进行了使用相同的方法作为KD experiments.Main结果和机会的作用：KD的Spin 1在MII期猪卵母细胞减少中期促进因子和促分裂原活化蛋白激酶的活动，导致自发的原核形成没有钙激活。然而，DNA损伤反应是由Spin 1过表达引发的，产生了检查点蛋白H2 A。X.此外，Spin 1过表达阻断了中期-后期转换，并导致卵母细胞和胚胎中的多核化。大规模数据：无限制，预防的原因：本研究是基于SpinI异常表达水平的体外研究。这可能会或可能不会准确地反映在vivo.WIDER影响的情况下发现：SpinI是必不可少的维持MII逮捕，但高水平的Spin 1诱导卵母细胞和胚胎的DNA损伤。因此，精确调节Spin 1表达的系统在猪MII期卵母细胞和胚胎中起作用。
STUDY QUESTION: What is the function of Spindlin I (Spin I) in metaphase II stage oocytes in pigs?SUMMARY ANSWER: Depletion of Spin I induces spontaneous oocyte activation and overexpression of SpinI causes multinuclear formation through induction of DNA damage in porcine oocytes.WHAT IS KNOWN ALREADY: Little is known about the function of Spin1 in oocytes and embryos. In mouse oocytes, Spin1 is specifically expressed during gametogenesis and is essential for meiotic resumption. In somatic cells, SpinI promotes cancer cell proliferation and activates WNT/T-cell factor signaling.STUDY DESIGN SIZE, DURATION: After knockdown (KD) or overexpression of SpinI in porcine MII-stage oocytes, MII maintenance was checked following additional culture for 24 h. Investigated parthenotes were cultured up to the four cell (72 h) or blastocyst (7 days) stages.PARTICIPANTS/MATERIALS, SETTING, METHODS: SpinI was knocked down in porcine oocytes and embryos via microinjection of pig SpinI-targeting siRNA. For Spin1 overexpression, porcine Spin l-eGFP cRNA was generated. Additionally, for rescue experiments, cRNA encoding siRNA-resistant mouse SpinI was added to the pig SpinI-targeting siRNA. For the overexpression and rescue experiments, microinjection and culture were performed using the same methods as the KD experiments.MAIN RESULTS AND THE ROLE OF CHANCE: KD of Spin1 in MII-stage porcine oocytes reduced metaphase-promoting factor and mitogen-activated protein kinase activities, resulting in spontaneous pronuclear formation without calcium activation. However, the DNA damage response was triggered by Spin1 overexpression, generating the checkpoint protein.H2A. X. Furthermore, Spin1 overexpression blocked metaphase-anaphase transition and led to multinucleation in oocytes and embryos.LARGE SCALE DATA: None.LIMITATIONS, REASONS FOR CAUTION: This study is based on in vitro investigations with abnormal expression levels of SpinI. This may or may not accurately reflect the situation in vivo.WIDER IMPLICATIONS OF THE FINDINGS: SpinI is essential to maintain MII arrest, but a high level of Spin1 induces DNA damage in oocytes and embryos. Thus, a system to accurately regulate Spin1 expression operates in porcine MII-stage oocytes and embryos.