与核重塑一样，胞质骨架重塑是受精卵和体细胞克隆胚发育中一个重要的生物学事件，对胚胎发育潜能具有重要影响。课题组前期研究发现牛精子来源的miR-202通过其靶基因SEPT7影响胚胎骨架蛋白修饰和胚胎细胞骨架重塑，但其影响机制和有关调控目前尚不清楚，为此，本项目拟通过对正常受精胚胎，采取功能缺失策略研究miR-202对其胞质骨架蛋白修饰、关键骨架结构（纺锤体、中心体和分裂收缩环）重塑及发育的影响；通过对缺少精子来源miRNA的核移植胚胎采取功能获得策略研究miR-202对牛体细胞克隆胚关键胞质骨架结构形成及胚胎发育的影响；并且通过Co-IP-LC-MS/MS和In Suit PLA等技术研究精子来源miR-202及其靶基因调控牛早期胚胎胞质骨架结构重塑过程的分子机制。该项目的开展将对哺乳动物胚胎发育调控以及核移植重编程机制的研究提供新的思路和科学依据。

As nuclear remodeling, cytoskeleton remodeling is a critical event during the early development of normal embryos and somatic cell nuclear transfer (SCNT) embryos, exerting significant influence on developmental potential of embryos. Our previous study indicated that bovine sperm-borne miR-202 could regulate the modifications of cytoskeletal proteins and cytoskeleton remodeling in embryos via targeting SEPT7. However, the underlying regulatory mechanisms are still largely unknown. To this end, this study would explore the effects of miR-202 on the modifications of cytoskeletal proteins, cytoskeleton (spindle, centrosome and contractile ring) remodeling, and embryonic development during preimplantation stage by loss-of- function. Gain-of-function would also be used to study its influence on cytoskeleton formation and embryonic development in bovine SCNT embryos that lack sperm-derived miRNAs. In addition, via utilizing techniques like Co-IP-LC-MS/MS and In Suit PLA, etc., we aim to demonstrate the molecular mechanisms of how sperm-borne miR-202 and its target gene regulate cytoskeleton remodeling in bovine early embryos. This study would provide new insights and scientific basis for mammalian embryo development regulation and the reprogramming mechanisms of SCNT embryos.