C. elegans Asymmetric Neuroblast Division

线虫不对称神经母细胞分裂

• 批准号: 6787279
• 负责人: GIAN GARRIGA
• 金额: $ 25.93万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-01 至 2006-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6787279
• 关键词: Caenorhabditis elegans; biological signal transduction; cell differentiation; cell growth regulation; developmental genetics; developmental neurobiology; gene expression; gene interaction; genetic regulation; genetic screening; immunocytochemistry; immunoprecipitation; invertebrate embryology; molecular asymmetry; molecular genetics; nerve stem cell; protein binding; protein localization; protein protein interaction; protein structure function

DESCRIPTION (provided by applicant): A fundamental question in development is how cells are specified. Elucidating mechanisms of cell-type specification is essential to the understanding and eventually treating human diseases, such as cancer, where this process is altered. One mechanism that specifies cell type is Asymmetric Cell Division, where a cell divides to produce two daughter cells that adopt distinct fates. Both asymmetrically distributed molecules and cell signaling appear to polarize dividing cells, but a connection between these two mechanisms has been lacking. A physical interaction between HAM-l, an asymmetrically distributed molecule, and DSH-2, a conserved Wnt signaling component, provides a link between asymmetric molecules and signaling. The overall goal of the proposed research is to define the roles of DSH-2 and HAM-1 in asymmetric cell division. The proposal has four specific aims. 1. To explore the interaction between DSH-2 and HAM-1. Immunoprecipitation experiments will be conducted to confirm a DSH-2/HAM-1 complex. Molecular genetic experiments will probe whether the interaction between the proteins is necessary for their function in asymmetric divisions and whether one protein is necessary for the others distribution. 2. To test whether the asymmetric distributions of DSH-2 and HAM-1 are necessary for their roles in asymmetric cell division. The localization domains of both proteins will be defined, and mislocalization experiments will test whether asymmetric distribution of two proteins is important to specify daughter cell fate. 3. To determine which proteins act upstream of DSH-2 and HAM-1 to control their asymmetric distribution. Using genetic and immunocytochemical approaches, Wnts and Frizzled receptors will be tested for roles in the distribution of DSH-2 and HAM-1. These molecules will then be misexpressed to test whether asymmetric signaling regulates DSH-2 and HAM-1 distribution. 4. To determine how DSH-2 and HAM-1 signal to control asymmetric cell division. Using reverse genetic approaches, candidate genes will be tested for roles in asymmetric divisions that require DSH-2 and HAM-1. In addition, genes identified in genetic screens will be tested for a role the HAM-1 /DSH-2 pathway. Genes that act in the pathway will be characterized molecularly.

描述（由申请人提供）：开发中的一个基本问题是 细胞是如何被指定的。阐明细胞类型规范的机制是 对于理解和最终治疗人类疾病至关重要，例如 癌症，这个过程被改变了。一种指定细胞类型的机制 是不对称细胞分裂，一个细胞分裂产生两个子细胞 有着截然不同的命运不对称分布的分子和细胞 信号似乎阻止分裂细胞，但这两者之间的联系 机制是缺乏的。HAM-1与HAM-1之间的物理相互作用 不对称分布的分子，和DSH-2，一个保守的Wnt信号转导分子， 它提供了不对称分子和信号传导之间的联系。的 所提出的研究的总体目标是确定DSH-2和HAM-1的作用 不对称细胞分裂该提案有四个具体目标。 1.探讨DSH-2与HAM-1的相互作用。免疫沉淀 将进行实验以确认DSH-2/HAM-1复合物。分子 遗传实验将探索蛋白质之间的相互作用是否是 它们在不对称分裂中的功能所必需的，以及一种蛋白质是否 其他的分配。 2.为了测试DSH-2和HAM-1的不对称分布是否 这是它们在不对称细胞分裂中所必需的。本地化领域 这两种蛋白质将被定义，和错误定位实验将测试 两种蛋白质的不对称分布是否重要， 子细胞的命运 3.为了确定哪些蛋白质作用于DSH-2和HAM-1的上游以控制它们的表达， 不对称分布利用遗传学和免疫细胞化学方法， 和Frizzled受体将被测试在DSH-2分布中的作用 HAM-1。然后这些分子将被错误表达，以测试是否不对称 信号传导调节DSH-2和HAM-1分布。 4.确定DSH-2和HAM-1信号如何控制细胞不对称分裂。 利用反向遗传学方法，将测试候选基因在以下方面的作用： 不对称分区需要DSH-2和HAM-1。此外，基因 将测试HAM-1 /DSH-2在基因筛选中的作用， 通路在该途径中起作用的基因将被分子表征。