Gap Junction and cAMP-Dependent Protein Kinase

9514285 Murray体外方法将用于研究间隙连接和基因调控后，已知的化合物影响连接表达和促进肾上腺甾体生成的细胞。特别地，将研究促肾上腺皮质激素（ACTH）和DbcAMP治疗以及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培养物。将含有针对间隙连接基因产物（Connexin 43, Cx43）的cDNA反义载体导入细胞，以确定间隙连接表达抑制的效果。将研究含有Cx43基因敲除的转基因小鼠的肾上腺发育。过表达和抑制间隙连接对肾上腺细胞功能的影响将被研究。通过免疫组织化学、western blot和northern blot分析来表征和定位间隙连接的表达。细胞通讯的证据将通过路西法黄色染料转移实验获得。对pKA和间隙连接表达的作用的研究将增加对间隙连接如何在内分泌细胞反应中起作用的认识。＊＊＊

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. ***