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Published online
doi:10.1083/jcb.200904140
The Journal of Cell Biology, Vol. 187, No. 3, 375-384
The Rockefeller University Press, 0021-9525 $30.00
© Loo et al.
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Heterogeneity in the physiological states and pharmacological responses of differentiating 3T3-L1 preadipocytes



Lit-Hsin Loo1,2, Hai-Jui Lin1,2, Dinesh K. Singh1,2, Kathleen M. Lyons1,2, Steven J. Altschuler1,2, and Lani F. Wu1,2

1 Green Center for Systems Biology and 2 Department of Pharmacology, University of Texas Southwestern Medical Center, Dallas, TX 75390

Correspondence to Steven J. Altschuler: steven.altschuler{at}utsouthwestern.edu; or Lani F. Wu: lani.wu{at}utsouthwestern.edu

Increases in key components of adipogenesis and lipolysis pathways correlate at the population-averaged level during adipogenesis. However, differentiating preadipocytes are highly heterogeneous in cellular and lipid droplet (LD) morphologies, and the degree to which individual cells follow population-averaged trends is unclear. In this study, we analyze the molecular heterogeneity of differentiating 3T3-L1 preadipocytes using immunofluorescence microscopy. Unexpectedly, we only observe a small percentage of cells with high simultaneous expression of markers for adipogenesis (peroxisome proliferator-activated receptor {gamma} [PPAR{gamma}], CCAAT/enhancer-binding protein {alpha}, and adiponectin) and lipid accumulation (hormone-sensitive lipase, perilipin A, and LDs). Instead, we identify subpopulations of cells with negatively correlated expressions of these readouts. Acute perturbation of adipocyte differentiation with PPAR{gamma} agonists, forskolin, and fatty acids induced subpopulation-specific effects, including redistribution of the percentage of cells in observed subpopulations and differential expression levels of PPAR{gamma}. Collectively, our results suggested that heterogeneity observed during 3T3-L1 adipogenesis reflects a dynamic mixture of subpopulations with distinct physiological states.

Abbreviations used in this paper: AdipoQ, adiponectin; C/EBP{alpha}, CCAAT/enhancer-binding protein {alpha}; FFA, free fatty acid; HSL, hormone-sensitive lipase; LD, lipid droplet; pHSL, phospho-HSL; PPAR{gamma}, peroxisome proliferator-activated receptor {gamma}.

© 2009 Loo et al.
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