Follicle-stimulating hormone-induced Gαh/phospholipase C-δ1 signaling mediating a noncapacitative Ca2+ influx through T-type Ca2+ channels in rat sertoli cells

Tsung Hsuan Lai, Yuan Feng Lin, Feng Chang Wu, Yu-Hui Tsai

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Our previous study demonstrated that FSH-induced immediate Ca2+ influx in rat Sertoli cells (SCs) is mediated by the Gαh/phospholipase C-δ1 (PLC-δ1) signaling pathway. As to which Ca2+ channel is responsible for such Ca2+ influx was not understood. In this study, thapsigargin triggered an instore calcium release and evoked a 1.5-fold elevation of intracellular Ca2+ in Ca2+-free media, whereas FSH exhibited no effect. The readdition of CaCl2 (2.5 mM) to FSH-pretreated or thapsigargin-sensitized SCs in Ca2+-free media immediately elicited a rapid Ca2+ influx or a 2-fold increase of second intracellular Ca2+ elevation, respectively. The addition of Ca2+ chelator EGTA (0.2 mM) reduced the FSH-induced elevation of intracellular Ca2+ in SCs incubated with CaCl2. However, pretreatment with dantrolene (25 μM), which inhibits in-store calcium release, did not affect the FSH-induced elevation of intracellular Ca 2+. NiCl2 (10 μM), a T-type calcium channel blocker, abolished the FSH-induced SC Ca2+ influx. Furthermore, mibefradil (10 and 100 μM), another specific blocker for T-type Ca2+ channels, dose-dependently suppressed the FSH-induced Ca2+ influx. In contrast, nifedipine (10 and 50 μM) or ω-conotoxin GVIA (100 and 500 nM), blocker of L- or N-type Ca2+ channels, respectively, did not affect the FSH-induced SC Ca2+ influx. On the other hand, FSH-induced Ca2+ influx was significantly reduced by pretreatment of SCs with myristoylated synthetic peptide (0.1 and 1 μM) of PLC-δ1 fragment TIPWNSLKQGYRHVHLL but not affected by 2′,5′-dideoxyadenosine (3 and 15 μM), a selective inhibitor of adenylate cyclase. In conclusion, the FSH-induced Gαh/PLC-δ1 pathway-dependent Ca2+ influx of rat SCs is mediated by T-type Ca2+ channels and independent of in-store calcium release.

Original languageEnglish
Pages (from-to)1031-1037
Number of pages7
JournalEndocrinology
Volume149
Issue number3
DOIs
Publication statusPublished - Mar 2008

Fingerprint

Sertoli Cells
Follicle Stimulating Hormone
Type C Phospholipases
Thapsigargin
Calcium
Dideoxyadenosine
Mibefradil
T-Type Calcium Channels
Conotoxins
Dantrolene
Egtazic Acid
Calcium Channel Blockers
Nifedipine
Chelating Agents
Peptides

ASJC Scopus subject areas

  • Endocrinology
  • Endocrinology, Diabetes and Metabolism

Cite this

Follicle-stimulating hormone-induced Gαh/phospholipase C-δ1 signaling mediating a noncapacitative Ca2+ influx through T-type Ca2+ channels in rat sertoli cells. / Lai, Tsung Hsuan; Lin, Yuan Feng; Wu, Feng Chang; Tsai, Yu-Hui.

In: Endocrinology, Vol. 149, No. 3, 03.2008, p. 1031-1037.

Research output: Contribution to journalArticle

@article{e292ecb40253488db1f33f9204f76a5d,
title = "Follicle-stimulating hormone-induced Gαh/phospholipase C-δ1 signaling mediating a noncapacitative Ca2+ influx through T-type Ca2+ channels in rat sertoli cells",
abstract = "Our previous study demonstrated that FSH-induced immediate Ca2+ influx in rat Sertoli cells (SCs) is mediated by the Gαh/phospholipase C-δ1 (PLC-δ1) signaling pathway. As to which Ca2+ channel is responsible for such Ca2+ influx was not understood. In this study, thapsigargin triggered an instore calcium release and evoked a 1.5-fold elevation of intracellular Ca2+ in Ca2+-free media, whereas FSH exhibited no effect. The readdition of CaCl2 (2.5 mM) to FSH-pretreated or thapsigargin-sensitized SCs in Ca2+-free media immediately elicited a rapid Ca2+ influx or a 2-fold increase of second intracellular Ca2+ elevation, respectively. The addition of Ca2+ chelator EGTA (0.2 mM) reduced the FSH-induced elevation of intracellular Ca2+ in SCs incubated with CaCl2. However, pretreatment with dantrolene (25 μM), which inhibits in-store calcium release, did not affect the FSH-induced elevation of intracellular Ca 2+. NiCl2 (10 μM), a T-type calcium channel blocker, abolished the FSH-induced SC Ca2+ influx. Furthermore, mibefradil (10 and 100 μM), another specific blocker for T-type Ca2+ channels, dose-dependently suppressed the FSH-induced Ca2+ influx. In contrast, nifedipine (10 and 50 μM) or ω-conotoxin GVIA (100 and 500 nM), blocker of L- or N-type Ca2+ channels, respectively, did not affect the FSH-induced SC Ca2+ influx. On the other hand, FSH-induced Ca2+ influx was significantly reduced by pretreatment of SCs with myristoylated synthetic peptide (0.1 and 1 μM) of PLC-δ1 fragment TIPWNSLKQGYRHVHLL but not affected by 2′,5′-dideoxyadenosine (3 and 15 μM), a selective inhibitor of adenylate cyclase. In conclusion, the FSH-induced Gαh/PLC-δ1 pathway-dependent Ca2+ influx of rat SCs is mediated by T-type Ca2+ channels and independent of in-store calcium release.",
author = "Lai, {Tsung Hsuan} and Lin, {Yuan Feng} and Wu, {Feng Chang} and Yu-Hui Tsai",
year = "2008",
month = "3",
doi = "10.1210/en.2007-1244",
language = "English",
volume = "149",
pages = "1031--1037",
journal = "Endocrinology",
issn = "0013-7227",
publisher = "The Endocrine Society",
number = "3",

}

TY - JOUR

T1 - Follicle-stimulating hormone-induced Gαh/phospholipase C-δ1 signaling mediating a noncapacitative Ca2+ influx through T-type Ca2+ channels in rat sertoli cells

AU - Lai, Tsung Hsuan

AU - Lin, Yuan Feng

AU - Wu, Feng Chang

AU - Tsai, Yu-Hui

PY - 2008/3

Y1 - 2008/3

N2 - Our previous study demonstrated that FSH-induced immediate Ca2+ influx in rat Sertoli cells (SCs) is mediated by the Gαh/phospholipase C-δ1 (PLC-δ1) signaling pathway. As to which Ca2+ channel is responsible for such Ca2+ influx was not understood. In this study, thapsigargin triggered an instore calcium release and evoked a 1.5-fold elevation of intracellular Ca2+ in Ca2+-free media, whereas FSH exhibited no effect. The readdition of CaCl2 (2.5 mM) to FSH-pretreated or thapsigargin-sensitized SCs in Ca2+-free media immediately elicited a rapid Ca2+ influx or a 2-fold increase of second intracellular Ca2+ elevation, respectively. The addition of Ca2+ chelator EGTA (0.2 mM) reduced the FSH-induced elevation of intracellular Ca2+ in SCs incubated with CaCl2. However, pretreatment with dantrolene (25 μM), which inhibits in-store calcium release, did not affect the FSH-induced elevation of intracellular Ca 2+. NiCl2 (10 μM), a T-type calcium channel blocker, abolished the FSH-induced SC Ca2+ influx. Furthermore, mibefradil (10 and 100 μM), another specific blocker for T-type Ca2+ channels, dose-dependently suppressed the FSH-induced Ca2+ influx. In contrast, nifedipine (10 and 50 μM) or ω-conotoxin GVIA (100 and 500 nM), blocker of L- or N-type Ca2+ channels, respectively, did not affect the FSH-induced SC Ca2+ influx. On the other hand, FSH-induced Ca2+ influx was significantly reduced by pretreatment of SCs with myristoylated synthetic peptide (0.1 and 1 μM) of PLC-δ1 fragment TIPWNSLKQGYRHVHLL but not affected by 2′,5′-dideoxyadenosine (3 and 15 μM), a selective inhibitor of adenylate cyclase. In conclusion, the FSH-induced Gαh/PLC-δ1 pathway-dependent Ca2+ influx of rat SCs is mediated by T-type Ca2+ channels and independent of in-store calcium release.

AB - Our previous study demonstrated that FSH-induced immediate Ca2+ influx in rat Sertoli cells (SCs) is mediated by the Gαh/phospholipase C-δ1 (PLC-δ1) signaling pathway. As to which Ca2+ channel is responsible for such Ca2+ influx was not understood. In this study, thapsigargin triggered an instore calcium release and evoked a 1.5-fold elevation of intracellular Ca2+ in Ca2+-free media, whereas FSH exhibited no effect. The readdition of CaCl2 (2.5 mM) to FSH-pretreated or thapsigargin-sensitized SCs in Ca2+-free media immediately elicited a rapid Ca2+ influx or a 2-fold increase of second intracellular Ca2+ elevation, respectively. The addition of Ca2+ chelator EGTA (0.2 mM) reduced the FSH-induced elevation of intracellular Ca2+ in SCs incubated with CaCl2. However, pretreatment with dantrolene (25 μM), which inhibits in-store calcium release, did not affect the FSH-induced elevation of intracellular Ca 2+. NiCl2 (10 μM), a T-type calcium channel blocker, abolished the FSH-induced SC Ca2+ influx. Furthermore, mibefradil (10 and 100 μM), another specific blocker for T-type Ca2+ channels, dose-dependently suppressed the FSH-induced Ca2+ influx. In contrast, nifedipine (10 and 50 μM) or ω-conotoxin GVIA (100 and 500 nM), blocker of L- or N-type Ca2+ channels, respectively, did not affect the FSH-induced SC Ca2+ influx. On the other hand, FSH-induced Ca2+ influx was significantly reduced by pretreatment of SCs with myristoylated synthetic peptide (0.1 and 1 μM) of PLC-δ1 fragment TIPWNSLKQGYRHVHLL but not affected by 2′,5′-dideoxyadenosine (3 and 15 μM), a selective inhibitor of adenylate cyclase. In conclusion, the FSH-induced Gαh/PLC-δ1 pathway-dependent Ca2+ influx of rat SCs is mediated by T-type Ca2+ channels and independent of in-store calcium release.

UR - http://www.scopus.com/inward/record.url?scp=40849145703&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=40849145703&partnerID=8YFLogxK

U2 - 10.1210/en.2007-1244

DO - 10.1210/en.2007-1244

M3 - Article

VL - 149

SP - 1031

EP - 1037

JO - Endocrinology

JF - Endocrinology

SN - 0013-7227

IS - 3

ER -