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Calcium Ion as a Second Messenger for o-Nitrophenylsulfenyl-Adrenocorticotropin (NPS-ACTH) and ACTH in Bovine Adrenal Steroidogenesis¹

Faculty of Integrated Arts and Sciences (T.Y., K.H., S.K.), Hiroshima University, Higashihiroshima 739, Japan; Department of Biophysics and Life Sciences (T.K., Y.O., S.K.), Graduate School of Arts and Sciences, University of Tokyo at Komaba, Meguro, Tokyo 153, Japan

Address all correspondence and requests for reprints to: Takeshi Yamazaki or Shiro Kominami, Faculty of Integrated Arts and Sciences, Hiroshima University, Higashihiroshima 739, Japan. E-mail: takey{at}ipc.hiroshima-u.ac.jp

o-Nitrophenyl sulfenyl-modified ACTH (NPS-ACTH) stimulated steroidogenesis acutely in bovine fasciculata-reticularis cells without increase in cellular cAMP synthesis. Application of NPS-ACTH to the cultured cells induced Ca²⁺ signals in individual cells as detected by video-enhanced microscopic fluorescence measurements. The percentage of Ca²⁺ signaling cells corresponded well with the increase of steroidogenesis induced by NPS-ACTH below 1 nM. Treatment of the cells with nicardipine, a Ca²⁺ channel blocker, suppressed the Ca²⁺ signals except for the transient increase just after the addition of NPS-ACTH and also blocked completely the stimulative effect on the steroidogenesis of NPS-ACTH below 1 nM. At a dosage of NPS-ACTH higher than 10 nM, the stimulative effect of steroidogenesis was partly suppressed by nicardipine and also by AA-861, a lipoxygenase inhibitor. The action of NPS-ACTH might be mediated by both Ca²⁺ and lipoxygenase metabolite(s) of arachidonic acid as dual second messengers. The effect of ACTH in pM range on the steroidogenesis was suppressed completely by the treatment with nicardipine and AA-861 at the same time, indicating that the action was mediated by both Ca²⁺ and the lipoxygenase metabolite(s) but not by cAMP. cAMP plays a significant role as a second messenger for ACTH action only at ACTH concentrations greater than 10 pM.

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