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A Novel Follicle-Stimulating Hormone-Induced Gh/Phospholipase C-1 Signaling Pathway Mediating Rat Sertoli Cell Ca²⁺-Influx

Graduate Institute of Pharmaceutical Science (Y.-F.L., Y.-W.W., Y.-H.T.), Graduate Institute of Cell and Molecular Biology (M.-J.T., H.-L.H., Y.-H.T.), and Department of Physiology (Y.-H.L.), Medical School, Taipei Medical University, Taipei, Taiwan 110, Republic of China; and Department of Life Science (M.-J.T.), National Chung Cheng University, Chia-Yi, Taiwan 621, Republic of China

Address all correspondence and requests for reprints to: Yu-Hui Tsai, Ph.D., 250 Wu-Hsing Street, Taipei, Taiwan 110, Republic of China. E-mail: cmbyht18{at}tmu.edu.tw.

FSH is known to activate Gs/cAMP signaling pathway in Sertoli cells (SCs) to support spermatogenesis. However, the molecular mechanism of FSH-induced Gs/cAMP-independent Ca²⁺-influx in SCs is not clear. In this study, FSH indeed induced an immediate and dose-dependent intracellular Ca²⁺-elevation in rat SCs. In the presence of EDTA (2.5 mM) or in the absence of extracellular Ca²⁺, the FSH-induced intracellular Ca²⁺-elevation was abolished. The confocal microscopic observation of Ca²⁺ image revealed that the SC cellular Ca²⁺ level was gradually increased after 50 sec of FSH treatment. Dantrolene, a blocker of intracellular Ca²⁺ release, did not affect this FSH-induced intracellular Ca²⁺ elevation. The pretreatment of rat SCs with phosphatidylinositol-phospholipase C (PLC)-specific inhibitor, U73122 (3 and 10 µM), inhibited the FSH-induced Ca²⁺-influx in a dose-dependent manner, but treatment with Gs-specific inhibitor, NF449 (0.1 and 0.3 µM), did not. On the other hand, the activation of Gh was immediately induced by FSH in the rat SCs within 5 sec of treatment. The translocation of PLC-1 from cytosol to cell membrane and the formation of Gh /PLC-1 complexes occurred within 5 and 10 sec, respectively, of FSH exposure. The intracellular inositol 1,4,5-triphosphate (IP₃) production was also detected after 30 sec of FSH treatment. The synthetic peptide of PLC-1 (TIPWNSLKQGYRHVHLL), not Gs inhibitor, predominantly inhibited the FSH-induced PLC-1 translocation, formation of Gh /PLC-1 complex, intracellular IP₃ production, and Ca²⁺ influx. In contrast, the peptide did not interfere with FSH-induced intracellular cAMP accumulation. In conclusion, the FSH-induced immediate Ca²⁺ influx is unambiguously mediated by an alternative Gh /PLC-1/IP₃ pathway that is distinct from the Gs/cAMP pathway in rat SCs.

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