Hornomal Modification of Communication in Cultured Animal Cells (Biology)

培养动物细胞通讯的激素修饰（生物学）

• 批准号: 9450133
• 负责人: Mary Lee Ledbetter
• 金额: $ 11.77万
• 依托单位: The Scripps Research Institute
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-09-01 至 1996-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9450133&HistoricalAwards=false
• 关键词: Hornomal; Modification; Communication; Cultured; Animal

Dr. Mary Lee Ledbetter's previous work has used simple assays to measure the ability of cultured fibroblastic or epithelial cells to share small cytoplasmic molecules through gap junctions. This cooperation can be inhibited by treatment of the cells with the sodium-pump inhibitor ouabain. When kidney-derived epithelial cells are treated with the steroid hormone aldosterone, their ouabain inhibition is reduced. Recent work has suggested that calcium changes mediate the effects of ouabain and that aldosterone's action may involve an unusual mechanism independent of the cytoplasmic mineralocorticoid receptor. Tests will be carried out to determine whether ouabain and aldosterone also modify the expression an/or distribution of connexins, the gap junction protein, in cooperating cells. Experimental approaches include: connexin protein localization by immunofluorescent staining in intact cells; expression of connexin - specific RNA and protein and protein electrophoretic mobility in cell extracts; and rapid changes in functional communication, measured by dye injection. These studies will not only provide a more complete basis for understanding the interaction between ouabain, aldosterone and cell communication, they will also provide improved tools to study structure-function relationships in gap junction communication for future work. Interactive activities include participation in the core course for first-year graduate students at The Scripps Research Institute. This involves leading discussion sections, designing and implementing laboratory experiences, and offering occasional lectures in the course.

玛丽李莱德贝特博士以前的工作已经使用简单的测定来测量培养的成纤维细胞或上皮细胞通过间隙连接共享小细胞质分子的能力。 这种合作可以通过用钠泵抑制剂哇巴因处理细胞来抑制。 当用类固醇激素醛固酮处理肾源性上皮细胞时，它们的哇巴因抑制作用降低。 最近的研究表明，钙的变化介导哇巴因的作用，醛固酮的作用可能涉及一个不寻常的机制，独立于细胞质盐皮质激素受体。 将进行试验以确定哇巴因和醛固酮是否也改变协作细胞中连接蛋白（差距连接蛋白）的表达和/或分布。 实验方法包括：在完整细胞中通过免疫荧光染色进行的连接蛋白蛋白定位; 2细胞提取物中连接蛋白特异性RNA和蛋白质的表达以及蛋白质电泳迁移率; 3通过染料注射测量的功能通讯的快速变化。 这些研究不仅将为理解哇巴因、醛固酮和细胞通讯之间的相互作用提供更完整的基础，还将为未来的工作提供更好的工具来研究间隙连接通讯的结构-功能关系。 互动活动包括参加斯克里普斯研究所一年级研究生的核心课程。 这包括领导讨论部分，设计和实施实验室经验，并在课程中提供偶尔的讲座。