IDENTIFICATION OF THE GENE FOR THE ACTH RECEPTOR BY EXPRESSION CLONING

ADRENOCORTICOTROPIC HORMONE (ACTH) AND ITS RECEPTORS ARE ESSENTIAL MOLECULAR COMPONENTS IN THE PERCEPTION OF AND REACTION TO STRESS. STRESS-INDUCED DISEASES INCLUDE NUMEROUS NEUROLOGICAL DISORDERS, SUCH AS ALZHEIMER'S DISEASE, AND CARDIOVASCULAR AS WELL AS IMMUNOLOGICAL DISORDERS. ELUCIDATION OF THE STRUCTURE OF ACTH RECEPTORS IS IMPORTANT FOR UNDERSTANDING THE BIOCHEMICAL BASIS OF STRESS-INDUCED PATHOLOGIES. DURING THE PAST DECADE, NEUREX CORPORATION HAS DEVELOPED THETOOLS FOR THE SUCCESSFUL DETECTION, CHARACTERIZATION, AND IDENTIFICATION OF PHYSIOLOGICALLY RELEVANT ACTH RECEPTORS. IT IS NOW PLANNED TO ISOLATE THE GENES CODING FOR THE ACTH RECEPTORS FIRST FROM A RAT ADRENAL CDNA LIBRARY BY TRANSFECTING COS 7 CELLS WITH A RAT ADRENAL CDNA LIBRARY CONSTRUCTED IN THE EXPRESSION VECTOR PRC/CMV AND SELECTING CELLS EXPRESSING THE RECEPTOR BY A LIGAND-BINDING TECHNIQUE INVOLVING ANTIBODIES AGAINST THE CARBOXY TERMINAL OF ACTH. THE RAT ADRENAL ACTH RECEPTOR CDNA WILL THEN BE USED TO ISOLATE THE CORRESPONDING CDNA FOR THE HUMAN RECEPTOR FROM A HUMAN ADRENAL CDNA LIBRARY CONSTRUCTED IN THE VECTOR LAMBDA-UNIZAP-XR. THE HUMAN ACTH RECEPTOR AND SUBTYPES ISOLATED BY PCR CLONING WILL BE EXPRESSED STABLY IN CHO CELLS, AND THE BIOCHEMICAL COUPLING MECHANISMS WILL BE INVESTIGATED. THE DISTRIBUTION AND TISSUE-SPECIFIC EXPRESSION OF ACTH RECEPTOR AND SUBTYPES WILL BE STUDIED USING NORTHERN ANALYSIS, IN SITU HYBRIDIZATION, AND CYTOCHEMISTRY.