Determining the role of CDC14 during meiosis in mouse oocytes

描述(由申请人提供)：本提案的总体目标是确定双特异性磷酸酶CDC14在哺乳动物减数分裂中的作用。减数分裂是二倍体前体细胞产生单倍体的过程，它与产生配子的物种特有的分化程序有关。CDC14在整个进化过程中高度保守，是迄今为止研究过的所有生物体中一个关键的有丝分裂细胞周期调节因子。在萌芽酵母中，cdc14突变体不能正确地完成第一次减数分裂，在那里同系物被分离。减数分裂I的错误与人类的不分离和染色体异常有关。高等真核生物含有2种CDC14蛋白，即CDC14a和CDc14b，这两种蛋白的功能将在本提案中进行探讨。这项建议的具体目的是1)使用RNA干扰(RNAi)方法检验小鼠卵母细胞减数分裂所需的CDC14a和CDc14b的假说，2)检验CDc14a和CDc14b的过度表达将改变小鼠卵母细胞减数分裂的假说，并开发一种对活卵母细胞进行成像的方法，以及3)通过突变确定CDC14蛋白中调节其亚细胞定位的结构域。减数分裂是在雌性体内产生卵子，在雄性体内产生精子的过程。女性减数分裂在人类中非常容易出错，因为大约20%的卵子含有异常的染色体数量，当与精子受精时，会导致自发流产、死产，在活产中还会导致唐氏综合症等发育疾病。探索减数分裂细胞周期是如何调节的，是了解DNA分离中的错误如何与人类染色体疾病有关的关键。

DESCRIPTION (provided by applicant): The overall goal of this proposal is to determine the role of the dual-specificity phosphatase Cdc14 during mammalian meiosis. Meiosis is the process by which a diploid precursor cell produces haploids and it is linked to species-specific differentiation programs that generate gametes. Cdc14 is highly conserved throughout evolution and is a critical mitotic cell-cycle regulator in all organisms studied to date. In budding yeast, Cdc14 mutants are unable to properly complete the first meiotic division where homologs are segregated. Mistakes in meiosis I are linked to nondisjunctions and chromosomal anomalies in humans. Higher eukaryotes contain 2 Cdc14 proteins, Cdc14a and Cdc14b, and the functions of both will be explored in this proposal. The specific aims of this proposal are to 1) test the hypothesis that Cdc14a and Cdc14b are required for meiosis in the mouse oocyte using an RNA interference (RNAi) approach, 2) test the hypothesis that over-expression of Cdc14a and Cdc14b will alter meiosis in the mouse oocyte and develop a method to image living oocytes and 3) identify domains within the CDC14 proteins that regulate their subcellular localization during meiosis by mutagenesis. Meiosis is the process that generates eggs in females and sperm in males. Female meiosis is highly error- prone in humans as approximately 20% of all eggs contain abnormal chromosome numbers that, when fertilized by sperm, leads to spontaneous abortions, stillbirths and, in live births, developmental diseases like Down Syndrome. Exploring how the meiotic cell cycle is regulated is key to gaining an understanding of how errors in the segregation of DNA are linked to human chromosomal disorders.