J. Lipid Res. Neurobiology of Lipids (ISSN1683-5506)
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A more recent version of this article appeared on February 1, 2004

Papers In Press, published online ahead of print November 1, 2003
J. Lipid Res., doi:10.1194/jlr.M300314-JLR200
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Submitted on July 15, 2003
Revised on October 30, 2003
Accepted on October 31, 2003

Stimulation of cardiolipin biosynthesis by PPARalpha activation: Elevated CDP-diacylglycerol synthase activity is not required for the PPARalpha -mediated stimulation of cardiolipin biosynthesis in rat heart H9c2 cells

Jan J. Jiang, Biao Lu, Fred Y. Xu, Jennifer Gartshore, William A. Taylor, Andrew J. Halayko, Frank J. Gonzalez, Jun Takasaki, Patrick C. Choy, and Grant M. Hatch

Pharmacology and Therapeutics, University of Manitoba, Winnipeg, Manitoba R3E 0T6

Corresponding Author: hatchgm{at}ms.umanitoba.ca

The role of peroxisome proliferator-activated receptor a (PPARa) stimulated mitochondrial phospholipase A2 in cardiac cardiolipin (CL) biosynthesis was examined in an in vivo and in vitro model. Treatment of H9c2 rat heart embryonic cells with the PPARa agonist clofibrate increased the expression of the 14 kDa phospholipase A2 (PLA2) protein and enzyme activity in mitochondrial fractions but did not affect pool size of CL. Clofibrate treatment of H9c2 cells stimulated de novo CL biosynthesis from [1,3-3H]glycerol thus accounting for why the pool size of CL was unaltered when mitochondrial PLA2 activity was elevated. This was due to stimulation of CL biosynthesis through an increase in mitochondrial phosphatidylglycerolphosphate (PGP) synthase activity by clofibrate. The role of PPARa in stimulation of de novo CL biosynthesis in vivo was examined in mouse heart. Cardiac mitochondrial PLA2, PGP synthase and CDP-1,2-diacyl-sn-glycerol synthase (CDS-2) activities were elevated in mice fed clofibrate for 14 days compared to controls. In addition, CDS-2 mRNA levels were elevated indicating an increase in CDS-2 expression. In contrast to mouse heart, clofibrate-treatment did not affect the activity nor mRNA levels of CDS-2 in H9c2 cells indicating that the enzyme is regulated differently in rat heart embryonic H9c2 cells and mouse heart in vivo. In PPARa-null mice clofibrate feeding did not alter cardiac mitochondrial PLA2, PGP synthase and CDS-2 activities nor CDS-2 mRNA levels compared to untreated controls thus confirming that these enzymes are regulated by PPARa activation in vivo. These results clearly indicate that cardiac CL de novo biosynthesis may be regulated by PPARa-agonist in responsive rodent models but that activation of CDS-2 is not required for elevated de novo CL biosynthesis in cultured rat heart embryonic H9c2 cells. Hence, CDS-2 is an example of an enzyme that exhibits alternative regulation in vivo and in cultured cell lines.


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