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Phosphatidylinositol‐ and phosphatidylcholine‐dependent phospholipases C are involved in the mechanism of action of atrial natriuretic factor in cultured rat aortic smooth muscle cells
Author(s) -
Zannetti Antonella,
Luly Paolo,
Musanti Roberto,
Baldini Patrizia M.
Publication year - 1997
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/(sici)1097-4652(199703)170:3<272::aid-jcp8>3.0.co;2-l
Subject(s) - phospholipase c , phosphatidylinositol , protein kinase c , diacylglycerol kinase , phosphatidate , phosphoinositide phospholipase c , second messenger system , staurosporine , phospholipase d , phospholipase , phospholipase a2 , diglyceride , phospholipid , phosphatidylcholine , medicine , gq alpha subunit , phorbol , inositol , endocrinology , chemistry , biochemistry , biology , signal transduction , receptor , g protein , enzyme , membrane
We have investigated the involvement of specific phospholipase systems and their possible mutual relationship with the mechanism by which atrial natriuretic factor (ANF) increases phosphatidate (PA) and diacylglycerol (DAG) in rat aortic smooth muscle cells (RASMC), one of the major targets of this hormone. Our results indicate that ANF initially stimulates a phosphatidylinositol‐dependent phospholipase C (PI‐PLC) with a significant increase of DAG, enriched in arachidonate, and inositol trisphosphate (IP 3 ) and then a phosphatidylcholine‐dependent phospholipase C (PC‐PLC) with formation of DAG, enriched in myristate, and phosphocholine (Pcho). Moreover, ANF stimulates PA formation at an intermediate stage between early and late DAG formation. The transphosphatidylation reaction, as well as its labeling ratio, demonstrate that phosphatidylcholine‐dependent phospholipase D (PC‐PLD) is not involved. Our experiments with R59022, a DAG kinase (DAGK) inhibitor, indicate that such an increase may be due to the phosphorylation of DAG derived from phosphatidylinositol (PI) hydrolysis. Our results show that phorbol 12‐myristate 13 acetate (PMA) plays a significant role in late DAG formation and that Pcho is released concomitantly, suggesting there is a relationship between the two phospholipase Cs (PLCs) that occurs through a protein kinase C (PKC) translocation from cytosol to the plasma membrane. These findings are confirmed by the use of PKC inhibitors calphostin, H7, and staurosporine. The involvement of membrane phospholipid hydrolysis and the ensuing production of second messengers might explain the vasorelaxant effect of ANF. J. Cell. Physiol. 170:272–278, 1997. © 1997 Wiley‐Liss, Inc.