A novel follicle-stimulating hormone-induced Gαh/phospholipase C-δ1 signaling pathway mediating rat Sertoli cell Ca2+-influx

Yuan Feng Lin, Min Jen Tseng, Hui Ling Hsu, Yu Wei Wu, Yi-Hsuan Lee, Yu-Hui Tsai

Research output: Contribution to journalArticle

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Abstract

FSH is known to activate Gs/cAMP signaling pathway in Sertoli cells (SCs) to support spermatogenesis. However, the molecular mechanism of FSHinduced Gs/cAMP-independent Ca2+-influx in SCs is not clear. In this study, FSH indeed induced an immediate and dose-dependent intracellular Ca 2+-elevation in rat SCs. In the presence of EDTA (2.5 mM) or in the absence of extracellular Ca2+, the FSH-induced intracellular Ca 2+-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 Gαh was immediately induced by FSH in the rat SCs within 5 sec of treatment. The translocation of PLC-δ1 from cytosol to cell membrane and the formation of Gαh/PLC-δ1 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-δ1 (TIPWNSLKQGYRHVHLL), not Gs inhibitor, predominantly inhibited the FSH-induced PLC-δ1 translocation, formation of Gαh /PLC-δ1 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 Gαh /PLC-δ1/IP3 pathway that is distinct from the Gs/cAMP pathway in rat SCs.

Original languageEnglish
Pages (from-to)2514-2527
Number of pages14
JournalMolecular Endocrinology
Volume20
Issue number10
DOIs
Publication statusPublished - 2006

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Sertoli Cells
Follicle Stimulating Hormone
Type C Phospholipases
Dantrolene
Phosphoinositide Phospholipase C
Inositol 1,4,5-Trisphosphate
Spermatogenesis
Edetic Acid
Cytosol
Cell Membrane
Peptides

ASJC Scopus subject areas

  • Molecular Biology
  • Endocrinology, Diabetes and Metabolism

Cite this

A novel follicle-stimulating hormone-induced Gαh/phospholipase C-δ1 signaling pathway mediating rat Sertoli cell Ca2+-influx. / Lin, Yuan Feng; Tseng, Min Jen; Hsu, Hui Ling; Wu, Yu Wei; Lee, Yi-Hsuan; Tsai, Yu-Hui.

In: Molecular Endocrinology, Vol. 20, No. 10, 2006, p. 2514-2527.

Research output: Contribution to journalArticle

Lin, Yuan Feng ; Tseng, Min Jen ; Hsu, Hui Ling ; Wu, Yu Wei ; Lee, Yi-Hsuan ; Tsai, Yu-Hui. / A novel follicle-stimulating hormone-induced Gαh/phospholipase C-δ1 signaling pathway mediating rat Sertoli cell Ca2+-influx. In: Molecular Endocrinology. 2006 ; Vol. 20, No. 10. pp. 2514-2527.
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abstract = "FSH is known to activate Gs/cAMP signaling pathway in Sertoli cells (SCs) to support spermatogenesis. However, the molecular mechanism of FSHinduced Gs/cAMP-independent Ca2+-influx in SCs is not clear. In this study, FSH indeed induced an immediate and dose-dependent intracellular Ca 2+-elevation in rat SCs. In the presence of EDTA (2.5 mM) or in the absence of extracellular Ca2+, the FSH-induced intracellular Ca 2+-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 Gαh was immediately induced by FSH in the rat SCs within 5 sec of treatment. The translocation of PLC-δ1 from cytosol to cell membrane and the formation of Gαh/PLC-δ1 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-δ1 (TIPWNSLKQGYRHVHLL), not Gs inhibitor, predominantly inhibited the FSH-induced PLC-δ1 translocation, formation of Gαh /PLC-δ1 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 Gαh /PLC-δ1/IP3 pathway that is distinct from the Gs/cAMP pathway in rat SCs.",
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AU - Wu, Yu Wei

AU - Lee, Yi-Hsuan

AU - Tsai, Yu-Hui

PY - 2006

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AB - FSH is known to activate Gs/cAMP signaling pathway in Sertoli cells (SCs) to support spermatogenesis. However, the molecular mechanism of FSHinduced Gs/cAMP-independent Ca2+-influx in SCs is not clear. In this study, FSH indeed induced an immediate and dose-dependent intracellular Ca 2+-elevation in rat SCs. In the presence of EDTA (2.5 mM) or in the absence of extracellular Ca2+, the FSH-induced intracellular Ca 2+-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 Gαh was immediately induced by FSH in the rat SCs within 5 sec of treatment. The translocation of PLC-δ1 from cytosol to cell membrane and the formation of Gαh/PLC-δ1 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-δ1 (TIPWNSLKQGYRHVHLL), not Gs inhibitor, predominantly inhibited the FSH-induced PLC-δ1 translocation, formation of Gαh /PLC-δ1 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 Gαh /PLC-δ1/IP3 pathway that is distinct from the Gs/cAMP pathway in rat SCs.

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