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Anchoring cortical granules in the cortex ensures trafficking to the plasma membrane for post-fertilization exocytosis

机译:在皮质中固定皮质颗粒可确保转运至质膜以进行受精后的胞吐作用

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Following fertilization, cortical granules exocytose ovastacin, a metalloendopeptidase that cleaves ZP2 in the zona pellucida surrounding mouse eggs to prevent additional sperm binding. Using high- and super-resolution imaging with ovastacinsupmCherry/sup as a fluorescent marker, we characterize cortical granule dynamics at single granule resolution in transgenic mouse eggs. Newly-developed imaging protocols provide an unprecedented view of vesicular dynamics near the plasma membrane in mouse eggs. We discover that cortical granule anchoring in the cortex is dependent on maternal MATER and document that myosin IIA is required for biphasic trafficking to the plasma membrane. We observe local clearance of cortical actin during exocytosis and determine that pharmacologic or genetic disruption of trafficking to the plasma membrane impairs secretion of cortical granules and results in polyspermy. Thus, the regulation of cortical granule dynamics at the cortex-plasma membrane interface is critical for exocytosis and the post-fertilization block to sperm binding that ensures monospermic fertilization.
机译:受精后,皮质颗粒胞吐卵清蛋白,一种金属内肽酶,在小鼠卵子周围的透明带中切割ZP2,以防止其他精子结合。使用以ovastacin mCherry 为荧光标记物的高分辨率和超高分辨率成像,我们以单个颗粒分辨率表征转基因小鼠卵中的皮质颗粒动力学。新开发的成像协议为小鼠卵质膜附近的囊泡动力学提供了前所未有的视角。我们发现皮质颗粒锚定在皮质中取决于母体MATER,并证明肌球蛋白IIA是双相运输至质膜所必需的。我们观察到胞吐过程中皮质肌动蛋白的局部清除,并确定贩运至质膜的药理或遗传学破坏可损害皮质颗粒的分泌,并导致多精子症。因此,调节皮质-质膜界面处的皮质颗粒动力学对于胞吐作用和受精后与精子的结合(确保单精子受精)至关重要。

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