小鼠受精卵雄原核中组蛋白修饰相关重编程因子的研究

The terminally differentiated cells are reprogramed to pluripotent/totipotent in the cytoplasm during fertilization and somatic cell nuclear transfer in mammals. Epigenetic modification plays important roles during this process. Study on the reprogramming factors of zygotes will help us to understand the reprogramming mechanism during somatic cell nuclear transfer in mammals. Our previous results demonstrated that the parental pronuclei have asymmetric reprogramming capacities and that the reprogramming factors reside predominantly in the male pronucleus. In this study, we plan to identify critical histone modification related reprogramming factors in the paternal pronuclei by the proteomics analysis of the proteins of maternal pronuclei, paternal pronuclei and cytoplasm of mouse zygotes. The function of reprogramming factors candidates and the reprogramming mechanism during fertilization, somatic cell nuclear transfer and embryonic stem cell nuclear transfer will be further investigated. This study will help us to understand the molecular mechanism of early embryonic development and the reprogramming mechanism of somatic cell nuclear transfer in mammals, which will provide a theoretical basis for therapeutic cloning and regenerative medicine.

哺乳动物受精和体细胞核移植都是高度分化的细胞在卵母细胞胞质中发生重编程的过程，表观遗传学修饰在此过程中起着非常重要的作用。对受精卵中重编程因子的研究将有利于对体细胞核移植过程的重编程机制的理解。我们最新的研究成果证明了小鼠受精卵中雌雄原核具有不对称的重编程能力并且重编程因子在受精过程中有选择性地集中在雄原核中。本研究计划通过蛋白质谱分析明确小鼠受精卵的雌雄原核以及胞质中的蛋白组成，从在雄原核中高表达的与组蛋白修饰相关的蛋白中筛选出与重编程相关的因子，研究这些因子在小鼠受精胚胎、体细胞克隆胚胎以及胚胎干细胞克隆胚胎发育中的作用，以期获取促进体细胞重编程的重要因子，提高体细胞重编程的效率，探讨重编程因子对小鼠体细胞克隆胚胎发育的影响机制。本研究的开展不仅有利于体细胞克隆效率的提高，还有利于阐明哺乳动物早期胚胎发育的分子机制和体细胞克隆胚胎的重编程机制，为治疗性克隆和人类再生医学提供理论基础。

哺乳动物受精和体细胞核移植都是高度分化的细胞在卵母细胞胞质中发生重编程的过程，表观遗传学修饰在此过程中起着非常重要的作用。对受精卵中重编程因子的研究将有利于对体细胞核移植过程的重编程机制的理解。本研究通过蛋白质谱分析了小鼠受精卵的雌雄原核以及胞质中的蛋白组成，从雌雄原核差异表达的蛋白中筛选出候选蛋白，深入研究了候选蛋白在小鼠卵母细胞成熟、受精胚胎、体细胞克隆胚胎以及胚胎干细胞克隆胚胎发育中的作用。研究结果首次证明了NEK5在小鼠卵母细胞成熟和着床前胚胎发育过程中发挥了重要的作用。本研究的开展增强了对哺乳动物卵母细胞成熟机制、早期胚胎发育分子机制和细胞重编程机制的理解，为人类健康提供了理论基础。