Role of Gap Junctions in the Regulation of Cell Migration

间隙连接在细胞迁移调节中的作用

• 批准号: 0444398
• 负责人: Sandra Murray
• 金额: $ 11万
• 依托单位: University of Pittsburgh
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-07-01 至 2007-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0444398&HistoricalAwards=false
• 关键词: Role; Gap; Junctions; Regulation; Cell

This project incorporates cutting-edge techniques for the identification of gap junctions and measurements of cell migration. Several types of adrenal cells will be studied that exhibit a wide variation in gap junction expression, either because of the initial phenotype of the cell or through experimental stimulation/inhibition of second messengers within the cell. In these studies, the influence of altered surface gap junctions on the regulation of cell migration will be explored in depth. The hypothesis that gap junctions play a pivotal role in cell migration will be addressed. Migration will be studied in cultures of adrenal cells that either have or do not have gap junctions (Specific Aim I). Cells will be treated with specific cAMP antagonists, agonist or diluent, and their migration characteristics will be assessed. Migration will be assayed using the following techniques: (1) time lapse imaging of the nuclear displacement or cell leading edge displacement of cells prelabeled with membrane permeable dyes; (2) counting the number of cells moving from a wound edge into a denuded area; and (3) counting the number of cells migrating in a silicon template compartmentalization chamber, in a microfluidic chamber, or through a Boyden chamber filter. A combination of time-lapse video microscopic and immunocytochemical techniques will be used to identify and elucidate the role of gap junction dynamics in cell migration.Infrastructural Contributions/Broader ImpactThe laboratory will continue to serve as a resource for the training of undergraduate and graduate students, postdoctoral fellows, visiting professors from minority institutions, and for mentoring high school students and (shadow program) teachers. Past NSF research funding has provided for the exposure and training of students, postdoctoral fellows, and faculty in areas of cell biology, endocrinology and signal transduction. Through seminars and workshops presented both nationally and internationally, information has been, and will continue to be, provided to members of the scientific community from a number of disciplines. High school students will continue to develop research projects and present at city and state science fairs. Recently, two undergraduate students won first place awards at national meetings, and others will be encouraged to follow in their path.

该项目结合了用于识别缝隙连接和测量细胞迁移的尖端技术。将研究几种类型的肾上腺细胞，这些细胞在缝隙连接表达上表现出很大的差异，要么是因为细胞的初始表型，要么是因为实验上对细胞内第二信使的刺激/抑制。在这些研究中，将深入探讨改变表面缝隙连接对细胞迁移调控的影响。缝隙连接在细胞迁移中发挥关键作用的假设将得到解决。迁移将在有或没有缝隙连接的肾上腺细胞的培养中进行研究(特定目标I)。细胞将被特定的cAMP拮抗剂、激动剂或稀释剂处理，并将评估它们的迁移特性。迁移将使用以下技术进行分析：(1)预先用膜渗透染料标记的细胞的核位移或细胞前沿位移的时间推移成像；(2)计数从伤口边缘移动到剥离区域的细胞数量；以及(3)计算在硅模板隔室、微流室或通过Boyden室过滤器迁移的细胞数量。将结合延时视频显微镜和免疫细胞化学技术来识别和阐明缝隙连接动力学在细胞迁移中的作用。基础结构贡献/更广泛的影响该实验室将继续作为培训本科生和研究生、博士后研究员、少数族裔机构的客座教授以及指导高中生和(影子计划)教师的资源。过去NSF的研究资金用于细胞生物学、内分泌学和信号转导领域的学生、博士后研究员和教职员工的接触和培训。通过在国内和国际上举办的研讨会和讲习班，已经并将继续向来自多个学科的科学界成员提供信息。高中生将继续开展研究项目，并出席市和州的科学博览会。最近，两名本科生在全国会议上获得了一等奖，其他人将被鼓励追随他们的道路。