Basiri Marcus L, Blachon Stephanie, Chim Yiu-Cheung Frederick, Avidor-Reiss Tomer
Department of Biological Sciences, University of Toledo.
J Vis Exp. 2013 Sep 20(79):e50938. doi: 10.3791/50938.
Centrosomes are conserved microtubule-based organelles whose structure and function change dramatically throughout the cell cycle and cell differentiation. Centrosomes are essential to determine the cell division axis during mitosis and to nucleate cilia during interphase. The identity of the proteins that mediate these dynamic changes remains only partially known, and the function of many of the proteins that have been implicated in these processes is still rudimentary. Recent work has shown that Drosophila spermatogenesis provides a powerful system to identify new proteins critical for centrosome function and formation as well as to gain insight into the particular function of known players in centrosome-related processes. Drosophila is an established genetic model organism where mutants in centrosomal genes can be readily obtained and easily analyzed. Furthermore, recent advances in the sensitivity and resolution of light microscopy and the development of robust genetically tagged centrosomal markers have transformed the ability to use Drosophila testes as a simple and accessible model system to study centrosomes. This paper describes the use of genetically-tagged centrosomal markers to perform genetic screens for new centrosomal mutants and to gain insight into the specific function of newly identified genes.
中心体是基于微管的保守细胞器，其结构和功能在整个细胞周期和细胞分化过程中会发生显著变化。中心体对于在有丝分裂期间确定细胞分裂轴以及在间期形成纤毛至关重要。介导这些动态变化的蛋白质的身份仍仅部分为人所知，并且许多与这些过程相关的蛋白质的功能仍处于初级阶段。最近的研究表明，果蝇精子发生提供了一个强大的系统，可用于鉴定对中心体功能和形成至关重要的新蛋白质，以及深入了解已知参与者在中心体相关过程中的特定功能。果蝇是一种成熟的遗传模式生物，在其中可以很容易地获得并分析中心体基因的突变体。此外，光学显微镜的灵敏度和分辨率的最新进展以及强大的基因标记中心体标记物的开发，已经改变了将果蝇睾丸用作研究中心体的简单且可及的模型系统的能力。本文描述了使用基因标记的中心体标记物进行新的中心体突变体的遗传筛选，并深入了解新鉴定基因的特定功能。
