Oleate stimulation of incorporation of exogenous glycerol into cardiolipin in isolated perfused rat heart does not involve direct activation of the CDP-DG pathway |
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Authors: | SG Cao GM Hatch |
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Affiliation: | Department of Internal Medicine, University of Manitoba, Winnipeg, Canada. |
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Abstract: | Oleate has been shown previously to stimulate the in vitro activity of phosphatidylglycerol-phosphatase, an important enzyme in the cytidine-5'-diphosphate-1,2-diacyl-sn-glycerol pathway of phosphatidylglycerol and cardiolipin biosynthesis. In this study the in vivo effect of oleate on the biosynthesis of new phosphatidylglycerol and cardiolipin was investigated in the heart. Hearts were perfused for 60 min with Krebs-Henseleit buffer containing 1,3-3H]glycerol and 0.6 mM albumin in the absence or presence of 0.6 or 1.2 mM oleate. Total incorporation of radioactivity was higher in the oleate-treated hearts compared with controls and this was due to an exclusive incorporation of radioactive glycerol into the organic phase. Also, the radioactivity incorporated into phosphatidylglycerol and cardiolipin was higher in the oleate-treated hearts compared with controls; however, the increase was greater in hearts perfused with 0.6 mM oleate compared with 1.2 mM oleate, indicating that pathophysiological concentrations of oleate may attenuate the oleate-induced stimulation of glycerol incorporation into polyglycerophospholipids. The pool size of phosphatidylglycerol and cardiolipin were unchanged in oleate-perfused hearts compared with controls. To investigate if the biosynthesis of phosphatidylglycerol and cardiolipin via the cytidine-5'-diphosphate-1,2-diacyl-sn-glycerol pathway was authentically stimulated by oleate hearts were pulse labeled for 15 min with 0.1 mM 1,3-3H]glycerol and subsequently chased for 60 min with 0.1 mM glycerol in the absence or presence of 0.6 mM oleate in the perfusate. Radioactivity incorporated into phosphatidylglycerol and cardiolipin was unchanged compared with controls. Our data indicate that oleate increases the incorporation of exogenous glycerol into polyglycerophospholipids but not accelerate synthesis from prelabeled precursor pools. Accordingly, oleate does not appear to stimulate directly enzymes of the cytidine-5'-diphosphate-1,2-diacyl-sn-glycerol pathway in vivo. |
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