CAMP调控小鼠卵母细胞减数分裂的分子机制

High-fidelity chromosome is required for cell division in both mitosis and meiosis. Errors in chromosome segregation can lead to the generation of aneuploid eggs and infertility. Fertilization of aneuploid eggs is a major cause of pregnancy loss, and if there is survival to term, it will result in developmental disabilities. The accurate kinetochore–microtubule attachment is necessary for the chromosome segregation. Although numerous molecules have been proposed to contribute to separate chromosomes in oocyte meiosis, the underlying mechanisms remain to be discovered. In this study, we will investigate if the CAMP/C13orf8/ZNF828, a zinc-finger protein, is a novel regulator of chromosome separation. Using ICR mice were employed as the animal model. Taking advantage of in vitro culture, immunofluorescent staining, molecular cloning, siRNA injection and western blotting, we will investigate the possible role of CAMP inregulating the oocyte maturation.

细胞在进行有丝分裂或减数分裂时都依赖于高度精确的染色体分离来确保遗传物质的稳定性。在减数分裂过程中染色体分离错误会导致卵子成熟障碍和产生非整倍体卵子，通常伴随不育、流产、胎儿死亡及出生缺陷等生殖发育缺陷。而动粒-微管的正确连接是保证染色体精确分离的关键事件，目前已发现多个分子在该过程发挥重要作用，但有更多的未知因子及其调控机制仍不明确。在卵子减数分裂成熟过程中，本项目将研究证明锌指蛋白CAMP/C13orf8/ZNF828是一种新的调节染色体分离的因子，以小鼠为研究模型，通过卵子体外成熟体系、免疫荧光染色、siRNA注射、cRNA注射、蛋白免疫印迹等方法，深入研究CAMP调控小鼠卵母细胞减数分裂成熟的分子机制。

无论是有丝分裂还是减数分裂，细胞分裂时都需要高度精确的染色体分离来确保遗传物质的正确分配。如果染色体分离出现错误会导致卵子成熟障碍，产生非整倍体以及影响受精通常伴随不育、流产、胎儿死亡及出生缺陷等。而动粒-微管的正确连接是保证染色体精确分离的关键。目前已经发现许多分子在调节染色体分离时发挥着重要作用，但更多的未知因子及其调控机制仍不明确。最近在有丝分裂的细胞中发现一种调节动粒-微管连接的锌指蛋白，该蛋白被称作CAMP/C13orf8/ZNF828，但它在减数分裂中的功能还没有被研究。本项目以哺乳动物ICR小鼠为研究对象，通过细胞体外培养、免疫荧光染色、分子克隆、mRNA 注射、siRNA 注射、蛋白免疫印迹等方法，来研究CAMP对小鼠卵母细胞减数分裂成熟调节的分子机制。内分泌干扰物，干扰性行为，导致受精能力下降，本项目还研究了内分泌干扰物对卵子质量和受精率影响的机制。