Meiotic spindle pole formation in Drosophila females

雌性果蝇减数分裂纺锤体极的形成

• 批准号: 6562800
• 负责人: KIM S MCKIM
• 金额: $ 22.49万
• 依托单位: RUTGERS, THE STATE UNIV OF N.J.
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-01-01 至 2006-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6562800
• 关键词: Drosophilidae; aneuploidy; cell cycle; centrosome; chromosome movement; cytogenetics; egg /ovum; embryogenesis; female; female reproductive system; fluorescent in situ hybridization; gene mutation; green fluorescent proteins; immunocytochemistry; kinesin; meiosis; microtubules; mitotic spindle apparatus; mutant; protein localization; protein protein interaction; protein structure function; spindle pole body

DESCRIPTION (provided by applicant): During the first meiotic division, homologous chromosomes linked by chiasmata interact with the spindle microtubules and segregate to opposite poles. Defects in this process lead to aneuploidy in the fertilized egg and have serious consequences on development, often resulting in death of the developing embryo. In humans, aneuploidy is a leading cause of spontaneous abortions and infertility in women. If aneuploids do survive, they manifest with diseases such as Down's, Tumer's or Klinefelter's syndrome. In many organisms, including humans and flies, the female meiotic spindle lacks centrioles and the classical microtubule-organizing center at the poles. The centrosomes and their constituent proteins are usually thought to organize bipolar spindles and therefore, in female meiosis, spindle pole organization must occur by a novel mechanism. By utilizing the powerful genetic and cytological techniques available in studies involving Drosophila melanogaster females, it is our long-term goal to elucidate the mechanism by which acentrosomal meiotic spindles form and how they function to segregate the chromosomes. We have found that the subito (sub) gene is required for bipolar spindle formation and homolog segregation during female meiosis in Drosophila. The findings that sub encodes a kinesin-like motor protein and is required during early embryogenesis and male meiosis has stimulated several new experiments. The aims of this study are to identify the role of SUB in spindle formation and understand how the meiotic spindle forms and functions to segregate chromosomes in the absence of centrosomes. A combination of genetic, immunological and cytological techniques will be used to illuminate the functions of SUB. These studies will investigate three critical areas of spindle biology by: 1) determining the subcellular localization of the sub protein in meiotic spindles and characterize how SUB interacts with the microtubules, 2) determining what other proteins interacts with SUB, and 3) using live imaging techniques to investigate the mechanism of spindle pole formation through a real time analysis of meiosis and mitosis in wild-type and mutants. The results of these studies will provide insights into the mechanisms of spindle formation and homolog segregation during meiosis and mitosis.

描述（由申请人提供）：在第一次减数分裂期间，通过交叉连接的同源染色体与纺锤体微管相互作用并分离到相反的两极。这一过程中的缺陷导致受精卵的非整倍体，并对发育产生严重后果，通常导致发育中的胚胎死亡。在人类中，非整倍体是妇女自然流产和不孕的主要原因。如果非整倍体确实存活下来，它们会表现出唐氏综合征、Tumer综合征或Klinefelter综合征等疾病。在许多生物体中，包括人类和苍蝇，雌性减数分裂纺锤体缺乏中心粒和两极的经典微管组织中心。中心体和它们的组成蛋白通常被认为是组织双极纺锤体，因此，在雌性减数分裂中，纺锤体极组织必须通过一种新的机制发生。通过利用强大的遗传学和细胞学技术，涉及果蝇雌性的研究，这是我们的长期目标，阐明的机制，其中acentrosomal减数分裂纺锤体的形式和它们如何发挥作用，分离染色体。我们发现subito（sub）基因是果蝇雌性减数分裂过程中双极纺锤体形成和同源分离所必需的。亚编码一种类似驱动蛋白的运动蛋白，在早期胚胎发育和雄性减数分裂中是必需的，这一发现刺激了几项新的实验。本研究的目的是确定在纺锤体形成的作用，并了解如何减数分裂纺锤体的形式和功能，在没有中心体的染色体分离。遗传学、免疫学和细胞学技术的结合将被用来阐明这种基因的功能。这些研究将通过以下方式调查纺锤体生物学的三个关键领域：1）确定减数分裂纺锤体中亚蛋白的亚细胞定位，并表征纺锤体如何与微管相互作用，2）确定其他蛋白质与纺锤体相互作用，以及3）使用实时成像技术通过野生型和突变体中减数分裂和有丝分裂的真实的时间分析来调查纺锤体极形成的机制。这些研究结果将有助于深入了解减数分裂和有丝分裂过程中纺锤体形成和同源物分离的机制。