Chemical Regulation of MAGE Levels in Differentiating Adipocytes

Friday, October 28, 2011
Hall 1-2 (San Jose Convention Center)
Joseph Tillotson , Chaminade University of Honolulu, Honolulu, HI
Alan Saghatelian, PhD , Harvard University, Cambridge, MA
Edwin Homan , Harvard University, Cambridge, MA
Adipogenesis is the process of cell differentiation by which undifferentiated fibroblasts are stimulated to form mature adipocytes. Researchers have made compelling advances to understanding the regulation of differentiating adipocytes due to availability of in vitro models. The 3T3-L1 cell line has been one to be very useful in recognizing mechanisms of adipocyte differentiation. It was recently reported that the differentiation of 3T3-L1 cells from pre-adipocytes to mature adipocytes is promoted in part by a class of lipids called monoalkylglycerol ethers (MAGE). In addition, (S)-BEL, a phospholipase A2 inhibitor, has been found to decrease adipogenesis. We sought to test the hypothesis that MAGE contributes to cellular differentiation by depriving the cells of MAGE. We treated cells with the inhibitor (S)-BEL and introduced the monooxygenase enzyme that degrades this lipid. MAGE levels were measured by liquid chromatography-mass spectrometry (LC-MS), triglyceride accumulation was measured by Oil Red O staining, and protein expression was assessed by western blotting.