Gap Junction and cAMP-Dependent Protein Kinase

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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. ***