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Coordinate and Divergent Regulation of Corticotropin-Releasing Factor (CRF) and CRF-Binding Protein Expression in an Immortalized Amygdalar Neuronal Cell Line¹

Department of Psychiatry (J.J.M., A.R., J.W.K.) and Department of Surgery (S.S., A.B.), University of Cincinnati College of Medicine, and University of Cincinnati Neuroscience Program (A.B., J.W.K.), Cincinnati, Ohio 45267; and Cincinnati Veteran’s Affairs Medical Center (J.W.K.), Psychiatry Service, Cincinnati, Ohio 45220

Address all correspondence and requests for reprints to: Jeff Mulchahey, Ph.D., Department of Psychiatry, University of Cincinnati, College of Medicine, 231 Bethesda Avenue, P.O. Box 670559, Cincinnati, Ohio 45267-0559. E-mail: mulchajj{at}email.uc.edu

CRF is a 41-amino acid neuropeptide best known for its hypophysiotropic actions. CRF is widely distributed in the central nervous system in areas beyond the hypothalamus. CRF-binding protein (CRF-BP) regulates the bioavailability of CRF, and knowledge of the regulation of CRF-BP synthesis is an integral component of understanding the actions of CRF. To better study the regulation of CRF and CRF-BP, primary amygdalar cultures were immortalized by transfection with the SV 40 large T antigen. A clonal line that expresses CRF immunoreactivity and messenger RNA was selected. The production of CRF peptide and message by this line is regulated in a manner indistinguishable from primary cultures. We also observed that the immortalized cells express CRF-BP immunoreactivity and messenger RNA. The expression of both CRF and CRF-BP is positively regulated by forskolin and interleukin-6. Unlike CRF, the expression of CRF-BP message and peptide was increased by phorbol 12-myristate 13-acetate or dexamethasone. These results demonstrate that the synthesis of CRF and CRF-BP in this clonal cell line may be regulated in parallel by some agents but not by others. These data also suggest that dexamethasone may decrease the biological availability of CRF in the amygdala by increasing the expression of CRF-BP, rather than by decreasing CRF expression.

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