Journal
EXPERIMENTAL CELL RESEARCH
Volume 314, Issue 11-12, Pages 2249-2256Publisher
ELSEVIER INC
DOI: 10.1016/j.yexcr.2008.04.003
Keywords
leptin; regulated secretion; 3T3-L1 adipocytes; PI3 kinase independent; adipokines; insulin
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Funding
- NIDDK NIH HHS [R01 DK041096, R01 DK052852, DK041096, DK52852, R01 DK041096-18] Funding Source: Medline
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To better define the molecular mechanisms underlying leptin release from adipocytes, we developed a novel protocol that maximizes leptin production from 3T3-L1 adipocytes. The addition of a PPAR gamma agonist to the Isobutylmethylxanthine/Dexamethasone/Insulin differentiation cocktail increased leptin mRNA levels by 5-fold, maintained insulin sensitivity, and yielded mature phenotype in cultured adipocytes. Under these conditions, acute insulin stimulation for 2 h induced a two-fold increase in leptin secretion, which was independent of new protein synthesis, and was not due to alterations in glucose metabolism. Stimulation with insulin for 15 min induced the same level of leptin release and was blocked by Brefeldin A. Inhibiting PI3-kinase with wortmannin had no effect on insulin stimulation of leptin secretion. These studies show that insulin can stimulate leptin release via a PI3K independent mechanism and provide a cellular system for studying the effect of insulin and potentially other mediators on leptin secretion. (C) 2008 Published by Elsevier Inc.
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