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Monitoring dimeric status of IZUMO1 during the acrosome reaction in living spermatozoon

机译：监测生物体精子的肌体反应期间Izumo1的二聚体状态

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摘要

The acrosome reaction (AR) is indispensable for successful spermatozoon-oocyte fusion. Recent studies have indicated that sperm IZUMO1 gradually gathers in the equatorial segment (EQ), which is the initiation site of sperm-egg fusion, after the AR. In addition, by examining the binding process of oocytes and Izumol-expressing cultured cells to reconstitute the early steps of fertilization, we previously demonstrated that robust IZUMO1-dependent adhesion specifically occurs at the contact site along with the dimerization of IZUMO1. However, when IZUMO1 dimerizes after the AR in living spermatozoon is unknown. Here, we report dynamics of IZUMO1 dimerization during the AR in spermatozoa by combining transgenic mice and time-lapse imaging using a set of bimolecular fluorescence complementation (BiFC) probes. Surprisingly, dimeric IZUMO1 was already formed at the acrosomal cap region before the AR and redistributed into the EQ after the AR. We categorized the translocation of the dimer into two types: Type 1, the near-simultaneous appearance of BiFC signals with IZUMO1-mCherry; and Type 2, the delayed formation of dimer in the EQ. Those findings suggest that, before encountering oocytes, spermatozoa are prepared to boost their affinity with JUNO.
机译：取代反应（Ar）对于成功的精子 - 卵母细胞融合是必不可少的。最近的研究表明，在AR之后，精子Izumo1逐渐在赤道段（EQ）中聚集，这是精子蛋融合的起始位点。另外，通过检查卵母细胞和表达伊苏酚的培养细胞的结合过程，以重构施肥的早期步骤，我们之前证明了特异性地发生在接触部位的鲁棒Izumo1依赖性粘附以及Izumo1的二聚化。然而，当Izumo1在生物精子中的AR之后二胺时尚不清楚。这里，通过使用一组共分子荧光互补（BIFC）探针将转基因小鼠和延时成像组合通过结合转基因小鼠和延时成像来报告Izumo1二聚化的动态。令人惊讶的是，二聚体Izumo1已经形成在AR之前的拟囊体帽区，并在AR之后重新分布到方程中。我们将二聚体的易位分为两种类型：1型，近同时出现与Izumo1-MCHERRY的BIFC信号;和类型2，方程式中的二聚体的延迟形成。这些研究结果表明，在遇到卵母细胞之前，精子准备使他们与Juno的亲和力提升。

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