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异性分化程序有关。Cdc14在整个进化过程中高度保守，是迄今为止研究的所有生物体中关键的有丝分裂细胞周期调节因子。在出芽酵母中，Cdc14突变体不能正确完成同源物分离的第一次减数分裂。减数分裂I中的错误与人类的非分裂和染色体异常有关。高等真核生物含有2种Cdc14蛋白，Cdc14a和Cdc14b，本文将探讨这两种蛋白的功能。本研究的具体目的是：1)利用RNA干扰（RNAi）方法验证小鼠卵母细胞减数分裂需要Cdc14a和Cdc14b的假设；2)验证过表达Cdc14a和Cdc14b会改变小鼠卵母细胞减数分裂的假设，并开发一种对活卵母细胞成像的方法；3)通过诱变确定减数分裂过程中CDC14蛋白内调节其亚细胞定位的结构域。减数分裂是女性产生卵子，男性产生精子的过程。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，当与精子受精时，会导致自然流产、死产，在活产时，还会导致唐氏综合症等发育性疾病。探索减数分裂细胞周期是如何调控的，是了解DNA分离错误如何与人类染色体疾病联系在一起的关键。

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.