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Biology of Reproduction 67, 1218-1224 (2002)
© 2002 Society for the Study of Reproduction, Inc.

Regulation of Gonadotropin {alpha} Subunit Gene Expression by Dopamine D2 Receptor Agonist in Clonal Mouse Gonadotroph {alpha}T3-1 Cells1

Haruhiko Kanasaki2,a, Toshie Yoneharaa, Yoko Yamadaa, Kentaro Takahashia, Kohkichi Hataa, Rituto Fujiwakia, Hideyuki Yamamotob, Yusuke Takeuchib, Kohji Fukunagab, Eishichi Miyamotob, and Kohji Miyazakia

a Department of Obstetrics and Gynecology, Shimane Medical University, Izumo 693-8501, Japan b Department of Pharmacology, Kumamoto University School of Medicine, Kumamoto 860-0811, Japan

Pituitary prolactin biosynthesis is negatively regulated by hypothalamic dopamine through D2 receptors in pituitary lactotrophs, but little is known about the direct effect of dopamine on gonadotrophs. In this study, the clonal gonadotroph-derived cell line, {alpha}T3-1, was used to examine whether gene expression of the pituitary gonadotropin {alpha} subunit, stimulated with GnRH or pituitary adenylate cyclase-activating polypeptide (PACAP), was controlled by dopamine D2 receptor. Western blotting and reverse transcription-polymerase chain reaction analysis demonstrated the presence of dopamine D2 receptors in {alpha}T3-1 cells. Both GnRH and PACAP increased {alpha} subunit gene expression. GnRH-induced {alpha} subunit gene expression was not affected by quinpirol, a specific dopamine D2 receptor agonist. In contrast, PACAP-induced gene expression was significantly lower in the presence of quinpirol. The roles of extracellular signal-regulated kinase (ERK) and cAMP in the expression of the {alpha} subunit gene were examined. GnRH activated ERK, but PACAP did not, and the activation was not inhibited by quinpirol. GnRH-induced {alpha} subunit gene expression was completely inhibited by an ERK inhibitor, PD098059. Cyclic AMP accumulation in {alpha}T3-1 cells was increased by treatment with PACAP, and quinpirol inhibited this effect. GnRH did not affect cAMP production in these cells. These results suggest that in {alpha}T3-1 cells, dopamine D2 receptors negatively regulate pituitary {alpha} subunit gene expression in association with the cAMP-dependent pathway, but not with the ERK pathway.

1 This work was supported in part by Grants-in-Aid for Scientific Research from the Ministry of Education, Science, Sports, and Culture of Japan to H.K. and K.M. and by a grant from the Ministry of Health and Welfare to K.M.

2 Correspondence. FAX: 81-853-20-2264; kawa45{at}shimane-med.ac.jp




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