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The Journal of Neuroscience, November 12, 2003, 23(32):10231-10237

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Cellular/Molecular
Regulation of Vasopressin Gene Expression by cAMP and Glucocorticoids in Parvocellular Neurons of the Paraventricular Nucleus in Rat Hypothalamic Organotypic Cultures

Shinobu Kuwahara, Hiroshi Arima, Ryouichi Banno, Ikuko Sato, Noriko Kondo, and Yutaka Oiso

Department of Metabolic Diseases, Field of Internal Medicine, Nagoya University Graduate School of Medicine, Showa-ku, Nagoya 466-8550, Japan

Arginine vasopressin (AVP) in the parvocellular neurons of the paraventricular nucleus (PVN) is known to play an important role in the hypothalamo-pituitary-adrenal axis. In the present study, we examined how cAMP and glucocorticoids regulate AVP gene expression in the parvocellular neurons of the PVN in rat hypothalamic organotypic cultures with in situ hybridization. AVP heteronuclear (hn) RNA, an indicator for gene transcription, was induced in the PVN with incubation of forskolin as reported previously, and AVP mRNA was increased by forskolin in the presence of the gene transcription inhibitor 5,6-dichloro-1-D-ribofuranosylbenzimidazole (DRB). These data indicate that cAMP could increase not only gene transcription but also mRNA stability. Dexamethasone treatment, in contrast, significantly decreased AVP mRNA expression levels in the PVN, but this inhibitory action was abolished in the presence of DRB or the sodium channel blocker tetrodotoxin (TTX). However, when the hypothalamic slices were treated with forskolin, dexamethasone decreased AVP mRNA expression even in the presence of DRB and/or TTX. Furthermore, AVP hnRNA expression induced by forskolin was attenuated by dexamethasone treatment in the presence of TTX. These data indicate that dexamethasone could act on AVP cells independently of action potentials to decrease mRNA stability and to suppress AVP gene transcription during stimulation by cAMP. Thus, it was demonstrated that: (1) cAMP upregulates AVP gene transcriptionally and post-transcriptionally, (2) the mode of action of glucocorticoids was dependent on whether the cells were stimulated by cAMP, and (3) the interactions between cAMP and glucocorticoids encompass both gene transcription and mRNA stability.

Key words: vasopressin; cAMP; glucocorticoid; transcription; stress; hypothalamus

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Received May 23, 2003; revised September 6, 2003; accepted September 19, 2003.

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