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Angiotensin II induces phosphatidic acid formation in neonatal rat cardiac fibroblasts: Evaluation of the roles of phospholipases C and D

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Abstract

Phosphatidic acid has been proposed to contribute to the mitogenic actions of various growth factors. In32P-labeled neonatal rat cardiac fibroblasts, 100 nM [Sar1]angiotensin II was shown to rapidly induce formation of32P-phosphatidic acid. Levels peaked at 5 min (1.5-fold above control), but were partially sustained over 2 h. Phospholipase D contributed in part to phosphatidic acid formation, as32P- or3H-phosphatidylethanol was produced when cells labeled with [32P]H3PO4 or 1-O-[1,2-3H]hexadecyl-2-lyso-sn-glycero-3-phosphocholine were stimulated in the presence of 1% ethanol. [Sar1]angiotensin II-induced phospholipase D activity was transient and mainly mediated through protein kinase C (PKC), since PKC downregulation reduced phosphatidylethanol formation by 68%. Residual activity may have been due to increased intracellular Ca2+, as ionomycin also activated phospholipase D in PKC-depleted cells. Phospholipase D did not fully account for [Sar1]angiotensin II-induced phosphatidic acid: 1) compared to PMA, a potent activator of phospholipase D, [Sar1]angiotensin II produced more phosphatidic acid relative to phosphatidylethanol, and 2) PKC downregulation did not affect [Sar1]angiotensin II-induced phosphatidic acid formation. The diacylglycerol kinase inhibitor R59949 depressed [Sar1]angiotensin II-induced phosphatidic acid formation by only 21%, indicating that activation of a phospholipase C and diacylglycerol kinase also can not account for the bulk of phosphatidic acid. Thus, additional pathways not involving phospholipases C and D, such asde novo synthesis, may contribute to [Sar1]angiotensin II-induced phosphatidic acid in these cells. Finally, as previously shown for [Sar1]angiotensin II, phosphatidic acid stimulated mitogen activated protein (MAP) kinase activity. These results suggest that phosphatidic acid may function as an intracellular second messenger of angiotensin II in cardiac fibroblasts and may contribute to the mitogenic action of this hormone on these cells. (Mol Cell Biochem141: 135–143, 1994)

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Abbreviations

DAG:

diacylglycerol

DMSO:

dimethyl sulfoxide

lysoPC:

1-O-hexadecyl-2-lyso-sn-glycero-3-phosphocholine

NRCF:

newborn rat cardiac fibroblasts

PA:

phosphatidic acid

PAPase:

phosphatidic acid phosphohydrolase

PC:

phosphatidylcholine

PEt:

phosphatidylethanol

PI:

phosphatidylinositol

PL:

(labeled) phospholipids

PLC:

phospholipase C

PLD:

phospholipase D

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Authors and Affiliations

  1. Geisinger Clinic, Weis Center for Research, 26-11, 100 North Academy Avenue, 17822, Danville, PA, USA

    George W. Booz, Mohiuddin M. Taher, Kenneth M. Baker & Harold A. Singer

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  1. George W. Booz
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  2. Mohiuddin M. Taher
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  3. Kenneth M. Baker
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  4. Harold A. Singer
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Additional information

Drs. G. W. Booz and M. M. Taher contributed equally to the work described here.

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Booz, G.W., Taher, M.M., Baker, K.M. et al. Angiotensin II induces phosphatidic acid formation in neonatal rat cardiac fibroblasts: Evaluation of the roles of phospholipases C and D. Mol Cell Biochem 141, 135–143 (1994). https://doi.org/10.1007/BF00926177

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  • DOI: https://doi.org/10.1007/BF00926177

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