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Thematic Review |
Center for Human Nutrition and Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, TX; Department of Nutrition Sciences and Toxicology, University of California at Berkeley, Berkeley, CA; and Division of Endocrinology and Metabolism, Department of Medicine, University of California at San Francisco, San Francisco, CA
Published, JLR Papers in Press, June 1, 2006.
1 To whom correspondence should be addressed. e-mail: Elizabeth.Parks{at}utsouthwestern.edu
Isotopic measurement of biosynthetic rates of lipids in VLDL particles has long posed difficult technical problems. In this review, key methodologic issues and recent technical advances are discussed. A common problem for all biosynthetic measurements is the requirement to measure isotopic labeling of the true intracellular biosynthetic precursor pool. Two techniques that address this problem for lipid biosynthesis, and that are applicable to humans, have been developed—the combinatorial probability method (or mass isotopomer distribution analysis) and 2H2O incorporation. The theoretical basis and practical application of these methods, both of which involve mass spectrometry, are described. Issues relevant to specific lipid components of VLDL, such as differences in the labeling of the various particle lipids (phospholipid, cholesterol, etc.), and the contribution of an intrahepatic cytosolic triacylglycerol (TG) storage pool to VLDL-TG are discussed. In summary, advances in stable isotope-mass spectrometric techniques now permit accurate measurement of liver-TG synthesis and flux. In vivo regulation of the synthesis, assembly, and secretion of VLDL-TG in humans is thereby accessible to direct investigation. Patient-oriented research in conditions such as dyslipidemia and hepatic steatosis is made feasible by these scientific advances.
Supplementary key words liver metabolism • triglycerides • dietary fat • de novo lipogenesis
Abbreviations: DNL, de novo lipogenesis; ER, endoplasmic reticulum; FCR, fractional catabolic rate; MIDA, mass isotopomer distribution analysis; TG, triacylglycerols
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