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 如何指定单元格。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。一种指定细胞类型的机制 是不对称细胞分裂，细胞分裂产生两个子细胞 采取不同的命运。不对称分布的分子和细胞 信号传导似乎使分裂细胞极化，但这两者之间的联系 一直缺乏机制。 HAM-l 之间的物理相互作用 不对称分布的分子和保守的 Wnt 信号传导 DSH-2 成分，提供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。该提案有四个具体目标。 1. 探讨DSH-2与HAM-1之间的相互作用。免疫沉淀 将进行实验来确认 DSH-2/HAM-1 复合物。分子 基因实验将探究蛋白质之间的相互作用是否存在 对于它们在不对称分裂中的功能以及一种蛋白质是否是 其他分配所必需的。 2. 检验DSH-2和HAM-1的不对称分布是否 它们在不对称细胞分裂中的作用是必需的。本地化领域 两种蛋白质的定义将被定义，并且错误定位实验将测试 指定两种蛋白质的不对称分布是否重要 女儿细胞的命运。 3. 确定哪些蛋白质作用于 DSH-2 和 HAM-1 的上游来控制它们 不对称分布。使用遗传和免疫细胞化学方法，Wnts 将测试卷曲受体在 DSH-2 分布中的作用 和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 的分布。 4. 确定 DSH-2 和 HAM-1 如何发出信号来控制不对称细胞分裂。 使用反向遗传方法，将测试候选基因的作用 需要 DSH-2 和 HAM-1 的不对称分裂。此外，基因 基因筛选中鉴定出的 HAM-1 /DSH-2 的作用将被测试 途径。在此途径中发挥作用的基因将从分子角度进行表征。