Regulation of relaxin 3 gene expression via cAMP-PKA in a neuroblastoma cell line.
Relaxin 3 is expressed in neurons of the brain stem that inneravate wide areas of the forebrain. Relaxin 3 mRNA levels in these neurons are increased in response to restraint stress, and by central administration of corticotropin-releasing factor (CRF). In the present study, we observed that relaxin 3 was expressed in a mouse neuroblastoma cell line, Neuro2a, and investigated the intracellular signaling that activated relaxin 3 gene transcription in vitro. By means of a clone stably transfected with a relaxin 3 promoter-EGFP gene, we observed that dibutyryl cyclic AMP and forskolin increased the relaxin 3 promoter activity. These increases were inhibited by pretreatment with PKA inhibitors, H89 and KT5720. Moreover, the promoter activity was enhanced by CRF treatment after expression of CRF-R1 receptor on the cells. Taken together, these results indicate that relaxin 3 transcription is activated via the cAMP-PKA pathway in the downstream of CRF-R1.
Tanaka
Masaki
M
Department of Cell Biology, Research Institute for Neurological Diseases and Geriatrics, Kyoto Prefectural University of Medicine, Kyoto, Japan. mtanaka@koto.kpu-m.ac.jp
Watanabe
Yoshihisa
Y
Yoshimoto
Kanji
K
D000818:Animals; D001483:Base Sequence; D001933:Brain Stem / Q000378:metabolism; D003994:Bucladesine / Q000494:pharmacology*; D002227:Carbazoles / Q000494:pharmacology; D045744:Cell Line, Tumor; D005576:Colforsin / Q000494:pharmacology*; D017124:Conserved Sequence; D003346:Corticotropin-Releasing Hormone / Q000494:pharmacology; D015870:Gene Expression; D047908:Intracellular Signaling Peptides and Proteins / Q000494:pharmacology*; D007546:Isoquinolines / Q000494:pharmacology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000187:drug effects*; D011758:Pyrroles / Q000494:pharmacology; D018019:Receptors, Corticotropin-Releasing Hormone / Q000378:metabolism; D012065:Relaxin / Q000235:genetics / Q000378:metabolism*; D013449:Sulfonamides / Q000494:pharmacology; D000099052:CRF Receptor, Type 1
D002227:Carbazoles; D047908:Intracellular Signaling Peptides and Proteins; D007546:Isoquinolines; D011758:Pyrroles; D018019:Receptors, Corticotropin-Releasing Hormone; D013449:Sulfonamides; C015482:protein kinase modulator; D005576:Colforsin; C057416:KT 5720; D000099052:CRF Receptor, Type 1; D003994:Bucladesine; D012065:Relaxin; D003346:Corticotropin-Releasing Hormone; C063509:N-(2-(4-bromocinnamylamino)ethyl)-5-isoquinolinesulfonamide