{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,22]],"date-time":"2026-09-22T10:00:13Z","timestamp":1790071213176,"version":"4.0.1"},"reference-count":19,"publisher":"The Endocrine Society","issue":"12","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2004,12,1]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>GHRH plays a pivotal role in the regulation of both synthesis and secretion of GH in the anterior pituitary. In this study, we examined the molecular mechanism of depolarization-induced GHRH gene transcription using the hypothalamus cell line, Gsh+\/+, revealing the involvement of the transcription factor called nuclear factor of activated T cells (NFAT). GHRH, NFAT1, NFAT4, and related genes were endogenously expressed in Gsh+\/+ cells and the rat arcuate nucleus, where NFAT1 and GHRH were colocalized. Cellular excitation with high potassium potently stimulated endogenous GHRH gene 5\u2032-promoter activity as well as the NFAT-mediated gene transcription, the former being further enhanced by coexpression of NFAT. On the other hand, cyclosporin A (a calcineurin-NFAT inhibitor) or EGTA (a calcium chelator) significantly blocked the depolarization-induced GHRH gene transcription. EMSA and site-directed mutagenesis experiments showed the direct binding of NFAT at five sites of the GHRH promoter, among which the relative importance of three distal sites (\u2212417\/\u2212403, \u2212402\/\u2212387, \u2212317\/\u2212301) was suggested. Finally, elimination of all five sites completely abolished the NFAT-induced GHRH gene up-regulation. Altogether, our results suggest that the transcription factor NFAT is involved in the depolarization-induced transcriptional activation of GHRH gene in the neuronal cells.<\/jats:p>","DOI":"10.1210\/me.2003-0471","type":"journal-article","created":{"date-parts":[[2004,8,20]],"date-time":"2004-08-20T02:54:50Z","timestamp":1092970490000},"page":"3011-3019","source":"Crossref","is-referenced-by-count":22,"title":["Nuclear Factor of Activated T Cells (NFAT) Is Involved in the Depolarization-Induced Activation of Growth Hormone-Releasing Hormone Gene Transcription in Vitro"],"prefix":"10.1210","volume":"18","author":[{"given":"Masato","family":"Asai","sequence":"first","affiliation":[{"name":"Departments of Medicine (M.A., M.Y., N.M.-N., H.A., Y.O.), Nagoya 466-8550, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yasumasa","family":"Iwasaki","sequence":"additional","affiliation":[{"name":"Clinical Pathophysiology (Y.I., M.I.), Nagoya University Graduate School of Medicine and Hospital, Nagoya 466-8550, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Masanori","family":"Yoshida","sequence":"additional","affiliation":[{"name":"Departments of Medicine (M.A., M.Y., N.M.-N., H.A., Y.O.), Nagoya 466-8550, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Noriko","family":"Mutsuga-Nakayama","sequence":"additional","affiliation":[{"name":"Departments of Medicine (M.A., M.Y., N.M.-N., H.A., Y.O.), Nagoya 466-8550, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hiroshi","family":"Arima","sequence":"additional","affiliation":[{"name":"Departments of Medicine (M.A., M.Y., N.M.-N., H.A., Y.O.), Nagoya 466-8550, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Masafumi","family":"Ito","sequence":"additional","affiliation":[{"name":"Clinical Pathophysiology (Y.I., M.I.), Nagoya University Graduate School of Medicine and Hospital, Nagoya 466-8550, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Koji","family":"Takano","sequence":"additional","affiliation":[{"name":"Department of Nephrology and Endocrinology (K.T.), Faculty of Medicine, University of Tokyo, Tokyo 112-8655, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yutaka","family":"Oiso","sequence":"additional","affiliation":[{"name":"Departments of Medicine (M.A., M.Y., N.M.-N., H.A., Y.O.), Nagoya 466-8550, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"80","reference":[{"key":"2020071614152221900_R1","doi-asserted-by":"crossref","first-page":"714","DOI":"10.1002\/j.1460-2075.1996.tb00407.x","article-title":"Gsh-1, an orphan Hox gene, is required for normal pituitary development.","volume":"15","author":"Li","year":"1996","journal-title":"EMBO J"},{"key":"2020071614152221900_R2","doi-asserted-by":"crossref","first-page":"2149","DOI":"10.1210\/mend.15.12.0747","article-title":"Homeobox protein Gsh-1-dependent regulation of the rat GHRH gene promoter.","volume":"15","author":"Mutsuga","year":"2001","journal-title":"Mol Endocrinol"},{"key":"2020071614152221900_R3","doi-asserted-by":"crossref","first-page":"1693","DOI":"10.1210\/mend-3-11-1693","article-title":"Mouse growth-hormone-releasing hormone: precursor structure and expression in brain and placenta.","volume":"3","author":"Suhr","year":"1989","journal-title":"Mol Endocrinol"},{"key":"2020071614152221900_R4","doi-asserted-by":"crossref","first-page":"8749","DOI":"10.1073\/pnas.88.19.8749","article-title":"Expression of the rat growth hormone-releasing hormone gene in placenta is directed by an alternative promoter.","volume":"88","author":"Gonzalez-Crespo","year":"1991","journal-title":"Proc Natl Acad Sci USA"},{"key":"2020071614152221900_R5","doi-asserted-by":"crossref","first-page":"1502","DOI":"10.1210\/endo.136.4.7895659","article-title":"Presence of a spermatogenic-specific promoter in the rat growth hormone-releasing hormone gene.","volume":"136","author":"Srivastava","year":"1995","journal-title":"Endocrinology"},{"key":"2020071614152221900_R6","doi-asserted-by":"crossref","first-page":"64","DOI":"10.1006\/dbio.1999.9304","article-title":"Novel strategy yields candidate Gsh-1 homeobox gene targets using hypothalamus progenitor cell lines.","volume":"211","author":"Li","year":"1999","journal-title":"Dev Biol"},{"key":"2020071614152221900_R7","doi-asserted-by":"crossref","first-page":"707","DOI":"10.1146\/annurev.immunol.15.1.707","article-title":"Transcription factors of theNFAT family: regulation and function.","volume":"15","author":"Rao","year":"1997","journal-title":"Annu Rev Immunol"},{"key":"2020071614152221900_R8","doi-asserted-by":"crossref","first-page":"2300","DOI":"10.1128\/MCB.19.3.2300","article-title":"Requirement for transcription factor NFAT in interleukin-2 expression.","volume":"19","author":"Chow","year":"1998","journal-title":"Mol Cell Biol"},{"key":"2020071614152221900_R9","doi-asserted-by":"crossref","first-page":"28181","DOI":"10.1016\/S0021-9258(18)46911-6","article-title":"Expression of the transcription factor NFATp in a neuronal cell line and in the murine nervous system.","volume":"269","author":"Ho","year":"1994","journal-title":"J Biol Chem"},{"key":"2020071614152221900_R10","doi-asserted-by":"crossref","first-page":"1758","DOI":"10.1210\/mend.15.10.0702","article-title":"Regulation of insulin gene transcription by a Ca2+-responsive pathway involving calcineurin and nuclear factor of activated T cells.","volume":"15","author":"Lawrence","year":"2001","journal-title":"Mol Endocrinol"},{"key":"2020071614152221900_R11","doi-asserted-by":"crossref","first-page":"703","DOI":"10.1038\/44378","article-title":"L-type calcium channels and GSK-3 regulate the activity of NFATc4 in hippocampal neurons.","volume":"401","author":"Graef","year":"1999","journal-title":"Nature"},{"key":"2020071614152221900_R12","doi-asserted-by":"crossref","first-page":"182","DOI":"10.1038\/32419","article-title":"Role of the NFATc transcription factor in morphogenesis of cardiac valves and septum.","volume":"392","author":"de la Pompa","year":"1998","journal-title":"Nature"},{"key":"2020071614152221900_R13","doi-asserted-by":"crossref","first-page":"863","DOI":"10.1016\/S0092-8674(01)00396-8","article-title":"Signals transduced by Ca2+\/calcineurin and NFATc3\/c4 pattern the developing vasculature.","volume":"105","author":"Graef","year":"2001","journal-title":"Cell"},{"key":"2020071614152221900_R14","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1084\/jem.191.1.9","article-title":"The nuclear factor of activated T cells (NFAT) transcription factor NFATp (NFATc2) is a repressor of chondrogenesis.","volume":"191","author":"Ranger","year":"2000","journal-title":"J Exp Med"},{"key":"2020071614152221900_R15","doi-asserted-by":"crossref","first-page":"1473","DOI":"10.1016\/S0024-3205(99)00084-3","article-title":"The effect of cyclosporine administration on growth hormone release and serum concentrations of insulin-like growth factor-I in male rats.","volume":"64","author":"Lopez-Calderon","year":"1999","journal-title":"Life Sci"},{"key":"2020071614152221900_R16","doi-asserted-by":"crossref","first-page":"309","DOI":"10.1159\/000026350","article-title":"Cellular signaling mechanisms for stimulation of growth hormone secretion and growth hormone primary transcripts by immunosuppressant agents, FK506 and cyclosporin A, in cultured rat pituitary cells.","volume":"5","author":"Ohye","year":"1998","journal-title":"Neuroimmunomodulation"},{"key":"2020071614152221900_R17","doi-asserted-by":"crossref","first-page":"5266","DOI":"10.1210\/endo.138.12.5639","article-title":"Positive and negative regulation of the rat vasopressin gene promoter.","volume":"138","author":"Iwasaki","year":"1997","journal-title":"Endocrinology"},{"key":"2020071614152221900_R18","doi-asserted-by":"crossref","first-page":"2033","DOI":"10.1172\/JCI119008","article-title":"Corticotropin-releasing hormone excites adrenocorticotropin-secreting human pituitary adenoma cells by activating a nonselective cation current.","volume":"98","author":"Takano","year":"1996","journal-title":"J Clin Invest"},{"key":"2020071614152221900_R19","doi-asserted-by":"crossref","first-page":"2396","DOI":"10.1021\/bi00552a018","article-title":"New calcium indicators and buffers with high selectivity against magnesium and proteins: design, synthesis and properties of prototype structures.","volume":"19","author":"Tsien,","year":"1980","journal-title":"Biochemistry"}],"container-title":["Molecular Endocrinology"],"original-title":[],"language":"en","link":[{"URL":"http:\/\/academic.oup.com\/mend\/article-pdf\/18\/12\/3011\/8967306\/mend3011.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"http:\/\/academic.oup.com\/mend\/article-pdf\/18\/12\/3011\/8967306\/mend3011.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,6,25]],"date-time":"2021-06-25T03:51:47Z","timestamp":1624593107000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/mend\/article\/18\/12\/3011\/2741852"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2004,12,1]]},"references-count":19,"journal-issue":{"issue":"12","published-print":{"date-parts":[[2004,12,1]]}},"URL":"https:\/\/doi.org\/10.1210\/me.2003-0471","relation":{},"ISSN":["0888-8809","1944-9917"],"issn-type":[{"value":"0888-8809","type":"print"},{"value":"1944-9917","type":"electronic"}],"subject":[],"published":{"date-parts":[[2004,12,1]]}}}