Adenosine triphosphate-evoked cytosolic calcium oscillations in human granulosa-luteal cells: Role of protein kinase C

Chen Jei Tai, Sung Keun Kang, Peter C K Leung

研究成果: 雜誌貢獻文章

17 引文 (Scopus)

摘要

ATP has been shown to modulate progesterone production in human granulosa-luteal cells (hGLCs) in vitro. After binding to a G protein-coupled P2 purinergic receptor, ATP stimulates phospholipase C. The resultant production of diacylglycerol and inositol triphosphate activates protein kinase C (PKC) and intracellular calcium [Ca 2+] i mobilization, respectively. In the present study, we examined the potential cross-talk between the PKC and Ca 2+ pathway in ATP signal transduction. Specifically, the effect of PKC on regulating ATP-evoked [Ca 2+] i oscillations were examined in hGLCs. Using microspectrofluorimetry, [Ca 2+] i oscillations were detected in Fura-2 loaded hGLCs in primary culture. The amplitudes of the ATP-triggered [Ca 2+] i oscillations were reduced in a dose-dependent manner by pretreating the cells with various concentrations (1 nM to 10 μM) of the PKC activator, phorbol-12-myristate-13-acetate (PMA). A 10 μM concentration of PMA completely suppressed 10 μM ATP-induced oscillations. The inhibitory effect occurred even when PMA was given during the plateau phase of ATP evoked [Ca 2+] i oscillations, suggesting that extracellular calcium influx was inhibited. The role of PKC was further substantiated by the observation that, in the presence of a PKC inhibitor, bisindolylmaleimide I, ATP-induced [Ca 2+] i oscillations were not completely suppressed by PMA. Furthermore, homologous desensitization of ATP-induced calcium oscillations was partially reversed by bisindolylmaleimide I, suggesting that activated PKC may be involved in the mechanism of desensitization. These results demonstrate that PKC negatively regulates the ATP-evoked [Ca 2+] i mobilization from both intracellular stores and extracellular influx in hGLCs and further support a modulatory role of ATP and P2 purinoceptor in ovarian steroidogenesis.

原文英語
頁(從 - 到)773-777
頁數5
期刊Journal of Clinical Endocrinology and Metabolism
86
發行號2
DOIs
出版狀態已發佈 - 2001
對外發佈Yes

指紋

Luteal Cells
Calcium Signaling
Protein Kinase C
Adenosine Triphosphate
Calcium
Acetates
Purinergic P2 Receptors
Signal transduction
Protein C Inhibitor
Fura-2
Diglycerides
Type C Phospholipases
Inositol
Protein Kinase Inhibitors
GTP-Binding Proteins
Progesterone
Signal Transduction

ASJC Scopus subject areas

  • Biochemistry
  • Endocrinology, Diabetes and Metabolism

引用此文

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title = "Adenosine triphosphate-evoked cytosolic calcium oscillations in human granulosa-luteal cells: Role of protein kinase C",
abstract = "ATP has been shown to modulate progesterone production in human granulosa-luteal cells (hGLCs) in vitro. After binding to a G protein-coupled P2 purinergic receptor, ATP stimulates phospholipase C. The resultant production of diacylglycerol and inositol triphosphate activates protein kinase C (PKC) and intracellular calcium [Ca 2+] i mobilization, respectively. In the present study, we examined the potential cross-talk between the PKC and Ca 2+ pathway in ATP signal transduction. Specifically, the effect of PKC on regulating ATP-evoked [Ca 2+] i oscillations were examined in hGLCs. Using microspectrofluorimetry, [Ca 2+] i oscillations were detected in Fura-2 loaded hGLCs in primary culture. The amplitudes of the ATP-triggered [Ca 2+] i oscillations were reduced in a dose-dependent manner by pretreating the cells with various concentrations (1 nM to 10 μM) of the PKC activator, phorbol-12-myristate-13-acetate (PMA). A 10 μM concentration of PMA completely suppressed 10 μM ATP-induced oscillations. The inhibitory effect occurred even when PMA was given during the plateau phase of ATP evoked [Ca 2+] i oscillations, suggesting that extracellular calcium influx was inhibited. The role of PKC was further substantiated by the observation that, in the presence of a PKC inhibitor, bisindolylmaleimide I, ATP-induced [Ca 2+] i oscillations were not completely suppressed by PMA. Furthermore, homologous desensitization of ATP-induced calcium oscillations was partially reversed by bisindolylmaleimide I, suggesting that activated PKC may be involved in the mechanism of desensitization. These results demonstrate that PKC negatively regulates the ATP-evoked [Ca 2+] i mobilization from both intracellular stores and extracellular influx in hGLCs and further support a modulatory role of ATP and P2 purinoceptor in ovarian steroidogenesis.",
author = "Tai, {Chen Jei} and Kang, {Sung Keun} and Leung, {Peter C K}",
year = "2001",
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T1 - Adenosine triphosphate-evoked cytosolic calcium oscillations in human granulosa-luteal cells

T2 - Role of protein kinase C

AU - Tai, Chen Jei

AU - Kang, Sung Keun

AU - Leung, Peter C K

PY - 2001

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N2 - ATP has been shown to modulate progesterone production in human granulosa-luteal cells (hGLCs) in vitro. After binding to a G protein-coupled P2 purinergic receptor, ATP stimulates phospholipase C. The resultant production of diacylglycerol and inositol triphosphate activates protein kinase C (PKC) and intracellular calcium [Ca 2+] i mobilization, respectively. In the present study, we examined the potential cross-talk between the PKC and Ca 2+ pathway in ATP signal transduction. Specifically, the effect of PKC on regulating ATP-evoked [Ca 2+] i oscillations were examined in hGLCs. Using microspectrofluorimetry, [Ca 2+] i oscillations were detected in Fura-2 loaded hGLCs in primary culture. The amplitudes of the ATP-triggered [Ca 2+] i oscillations were reduced in a dose-dependent manner by pretreating the cells with various concentrations (1 nM to 10 μM) of the PKC activator, phorbol-12-myristate-13-acetate (PMA). A 10 μM concentration of PMA completely suppressed 10 μM ATP-induced oscillations. The inhibitory effect occurred even when PMA was given during the plateau phase of ATP evoked [Ca 2+] i oscillations, suggesting that extracellular calcium influx was inhibited. The role of PKC was further substantiated by the observation that, in the presence of a PKC inhibitor, bisindolylmaleimide I, ATP-induced [Ca 2+] i oscillations were not completely suppressed by PMA. Furthermore, homologous desensitization of ATP-induced calcium oscillations was partially reversed by bisindolylmaleimide I, suggesting that activated PKC may be involved in the mechanism of desensitization. These results demonstrate that PKC negatively regulates the ATP-evoked [Ca 2+] i mobilization from both intracellular stores and extracellular influx in hGLCs and further support a modulatory role of ATP and P2 purinoceptor in ovarian steroidogenesis.

AB - ATP has been shown to modulate progesterone production in human granulosa-luteal cells (hGLCs) in vitro. After binding to a G protein-coupled P2 purinergic receptor, ATP stimulates phospholipase C. The resultant production of diacylglycerol and inositol triphosphate activates protein kinase C (PKC) and intracellular calcium [Ca 2+] i mobilization, respectively. In the present study, we examined the potential cross-talk between the PKC and Ca 2+ pathway in ATP signal transduction. Specifically, the effect of PKC on regulating ATP-evoked [Ca 2+] i oscillations were examined in hGLCs. Using microspectrofluorimetry, [Ca 2+] i oscillations were detected in Fura-2 loaded hGLCs in primary culture. The amplitudes of the ATP-triggered [Ca 2+] i oscillations were reduced in a dose-dependent manner by pretreating the cells with various concentrations (1 nM to 10 μM) of the PKC activator, phorbol-12-myristate-13-acetate (PMA). A 10 μM concentration of PMA completely suppressed 10 μM ATP-induced oscillations. The inhibitory effect occurred even when PMA was given during the plateau phase of ATP evoked [Ca 2+] i oscillations, suggesting that extracellular calcium influx was inhibited. The role of PKC was further substantiated by the observation that, in the presence of a PKC inhibitor, bisindolylmaleimide I, ATP-induced [Ca 2+] i oscillations were not completely suppressed by PMA. Furthermore, homologous desensitization of ATP-induced calcium oscillations was partially reversed by bisindolylmaleimide I, suggesting that activated PKC may be involved in the mechanism of desensitization. These results demonstrate that PKC negatively regulates the ATP-evoked [Ca 2+] i mobilization from both intracellular stores and extracellular influx in hGLCs and further support a modulatory role of ATP and P2 purinoceptor in ovarian steroidogenesis.

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