Mechanism of phospholipase D activation induced by extracellular ATP in osteoblast-like cells

Atsushi Suzuki, J. Shinoda, Y. Oiso, O. Kozawa

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

We have previously reported that extracellular ATP stimulates Ca2+ influx from extracellular space, resulting in the production of prostaglandin E2 which mediates, at least in part, its proliferative effect on osteoblast-like MC3T3-E1 cells, and that the activation of protein kinase C (PKC) stimulates phospholipase D in these cells. In the present study, we examined the effect of extracellular ATP on phosphatidylcholine-hydrolysing phospholipase D activity in MC3T3-E1 cells. ATP stimulated the formation of both choline and inositol phosphates dose-dependently in the range between 0.1 and 1 mM. The formation of choline by a combination of ATP and NaF, an activator of GTP-binding protein, was synergistic, whereas that of inositol phosphates was not. A combination of ATP and 12-O-tetradecanoylphorbol-13-acetate, a PKC activating phorbol ester, additively stimulated the formation of choline. Staurosporine, an inhibitor of PKC, had little effect on ATP-stimulated formation of choline. Choline formation was significantly reduced by chelating extracellular Ca2+ with EGTA, while being inhibited by W-7, an antagonist of calmodulin. These results suggest that extracellular ATP stimulates phospholipase D in a Ca2+/calmodulin-dependent manner in osteoblast-like cells, and that neither PKC activation nor GTP-binding protein is involved in this mechanism.

Original languageEnglish
Pages (from-to)81-86
Number of pages6
JournalJournal of Endocrinology
Volume145
Issue number1
DOIs
Publication statusPublished - 01-01-1995
Externally publishedYes

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Phospholipase D
Osteoblasts
Adenosine Triphosphate
Choline
Protein Kinase C
Inositol Phosphates
Calmodulin
GTP-Binding Proteins
Staurosporine
Phosphorylcholine
Egtazic Acid
Extracellular Space
Phorbol Esters
Tetradecanoylphorbol Acetate
Phosphatidylcholines
Dinoprostone

All Science Journal Classification (ASJC) codes

  • Endocrinology, Diabetes and Metabolism
  • Endocrinology

Cite this

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abstract = "We have previously reported that extracellular ATP stimulates Ca2+ influx from extracellular space, resulting in the production of prostaglandin E2 which mediates, at least in part, its proliferative effect on osteoblast-like MC3T3-E1 cells, and that the activation of protein kinase C (PKC) stimulates phospholipase D in these cells. In the present study, we examined the effect of extracellular ATP on phosphatidylcholine-hydrolysing phospholipase D activity in MC3T3-E1 cells. ATP stimulated the formation of both choline and inositol phosphates dose-dependently in the range between 0.1 and 1 mM. The formation of choline by a combination of ATP and NaF, an activator of GTP-binding protein, was synergistic, whereas that of inositol phosphates was not. A combination of ATP and 12-O-tetradecanoylphorbol-13-acetate, a PKC activating phorbol ester, additively stimulated the formation of choline. Staurosporine, an inhibitor of PKC, had little effect on ATP-stimulated formation of choline. Choline formation was significantly reduced by chelating extracellular Ca2+ with EGTA, while being inhibited by W-7, an antagonist of calmodulin. These results suggest that extracellular ATP stimulates phospholipase D in a Ca2+/calmodulin-dependent manner in osteoblast-like cells, and that neither PKC activation nor GTP-binding protein is involved in this mechanism.",
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Mechanism of phospholipase D activation induced by extracellular ATP in osteoblast-like cells. / Suzuki, Atsushi; Shinoda, J.; Oiso, Y.; Kozawa, O.

In: Journal of Endocrinology, Vol. 145, No. 1, 01.01.1995, p. 81-86.

Research output: Contribution to journalArticle

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