Gap Junction and cAMP-Dependent Protein Kinase

间隙连接和 cAMP 依赖性蛋白激酶

• 批准号: 9514285
• 负责人: Sandra Murray
• 金额: $ 22.38万
• 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: 1996
• 资助国家: 美国
• 起止时间: 1996-07-01 至 2000-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9514285&HistoricalAwards=false
• 关键词: Gap; Junction; cAMP; Dependent; Protein

9514285 Murray In vitro methods will be used to study gap junctions and gene regulation following treatment of cells with compounds known to influence junction expression and to promote adrenal steroidogenesis. Specially, the effect of adrenocorticotropin (ACTH), and DbcAMP treatment and cAMP- dependent protein kinase (pKA) activity on gap junction expression will be studied. Bovine and human adrenal cortical cell lines and primary rat cultures of adult rat adrenal cortical cells will be used. Vectors containing cDNA antisense directed against gap junction gene products (Connexin 43, Cx43) will be introduced into cells to determine the effect of gap junction expression inhibition. Adrenal gland development in transgenic mice containing a Cx43 knockout will be studied. The effect of over expression and inhibition of gap junctions on adrenal cell function, will be studied. Gap junction expression will be characterized and localized with immunohistochemistry, western blot and northern blot analysis. Evidence of cell communication will be obtained with Lucifer yellow dye transfer experiments. Studies on the role of pKA and gap junction expression will increase the knowledge of how gap junctions function in endocrine cell response. ***

小行星9514285 体外方法将用于研究间隙连接和基因调控后的细胞与已知的影响连接表达和促进肾上腺类固醇生成的化合物的治疗。 特别地，将研究促肾上腺皮质激素（ACTH）和DbcAMP处理以及cAMP依赖性蛋白激酶（pKA）活性对间隙连接表达的影响。 将使用牛和人肾上腺皮质细胞系以及成年大鼠肾上腺皮质细胞的原代大鼠培养物。 将含有针对间隙连接基因产物（连接蛋白43，Cx43）的cDNA反义的载体引入细胞中以确定间隙连接表达抑制的效果。 将研究含有Cx43敲除的转基因小鼠中的肾上腺发育。 将研究间隙连接的过度表达和抑制对肾上腺细胞功能的影响。 缝隙连接表达将用免疫组织化学、蛋白质印迹和北方印迹分析表征和定位。 细胞通讯的证据将通过荧光黄染料转移实验获得。 对pKA和缝隙连接表达的作用的研究将增加对缝隙连接在内分泌细胞反应中的作用的认识。 ***