Heterogeneity in the physiological states and pharmacological responses of differentiating 3T3-L1 preadipocytes.
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 [PPARgamma], 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 PPARgamma agonists, forskolin, and fatty acids induced subpopulation-specific effects, including redistribution of the percentage of cells in observed subpopulations and differential expression levels of PPARgamma. Collectively, our results suggested that heterogeneity observed during 3T3-L1 adipogenesis reflects a dynamic mixture of subpopulations with distinct physiological states.
Loo
Lit-Hsin
LH
Green Center for Systems Biology, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA.
Lin
Hai-Jui
HJ
Singh
Dinesh K
DK
Lyons
Kathleen M
KM
Altschuler
Steven J
SJ
Wu
Lani F
LF
D017667:Adipocytes / Q000166:cytology / Q000187:drug effects; D050156:Adipogenesis / Q000502:physiology; D052242:Adiponectin / Q000378:metabolism; D000818:Animals; D023081:CCAAT-Binding Factor / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects*; D005576:Colforsin / Q000494:pharmacology; D005227:Fatty Acids / Q000494:pharmacology; D005819:Genetic Markers; D050356:Lipid Metabolism; D051379:Mice; D008856:Microscopy, Fluorescence; D041681:NIH 3T3 Cells; D047495:PPAR gamma / Q000819:agonists / Q000378:metabolism; D013997:Time Factors
D052242:Adiponectin; D023081:CCAAT-Binding Factor; D005227:Fatty Acids; D005819:Genetic Markers; D047495:PPAR gamma; D005576:Colforsin