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 如何指定细胞。阐明细胞类型规范的机制是 对理解和最终治疗人类疾病至关重要的人，例如 癌症，该过程发生了改变。一种指定单元类型的机制 是不对称的细胞分裂，其中一个细胞分裂以产生两个子细胞 那采取了不同的命运。不对称分布的分子和细胞都 信号传导似乎使分隔单元的偏振，但是这两个之间有连接 机制一直缺乏。 HAM-L之间的身体互动 不对称分布的分子和DSH-2，一种保守的Wnt信号传导 分量提供了不对称分子和信号传导之间的联系。这 拟议研究的总体目标是定义DSH-2和HAM-1的作用 在不对称细胞分裂中。该提案具有四个具体目标。 1。探索DSH-2和HAM-1之间的相互作用。免疫沉淀 将进行实验以确认DSH-2/HAM-1复合物。分子 基因实验将探测蛋白质之间的相互作用是否为 它们在非对称分裂中的功能所必需的以及一种蛋白质是否是 他人分配所需的。 2。测试DSH-2和HAM-1的不对称分布是否为 它们在非对称细胞分裂中的作用所必需的。本地化域 将定义这两种蛋白质，并将错误定位实验测试 两种蛋白质的不对称分布对于指定很重要 女儿细胞命运。 3。确定哪种蛋白在DSH-2和HAM-1的上游作用以控制其 不对称分布。使用遗传和免疫细胞化学方法，WNT 并将测试卷曲的受体在DSH-2的分布中的作用 和HAM-1。然后，这些分子将被压制以测试是否不对称 信号调节DSH-2和HAM-1分布。 4。确定DSH-2和HAM-1信号如何控制不对称细胞分裂。 使用反向遗传方法，将测试候选基因在 需要DSH-2和HAM-1的不对称分裂。另外，基因 在遗传筛选中确定的将测试HAM-1 /DSH-2的角色 路径。在途径中起作用的基因将被分子表征。