Abstract
Adipocytes have been fundamental in identifying basic cellular mechanisms associated with diabetes and obesity, but their bioenergetics has not been fully understood. We measured changes in oxygen consumption rates (OCR) and extracellular acidification rates (ECAR) during differentiation of naïve and TNF‐α exposed 3T3‐L1 preadipocytes using Seahorse XF24 Extracellular Flux Analyzer, and correlated them to mRNA and protein markers of adipogenesis. Basal OCR and ECAR increased as the 3T3‐L1 cells underwent adipogenic differentiation from 110.7±26.5 to 901.1±86.9 pmol/min, and from 7.7±1.4 to 21.6±1.8 mpH/min, respectively. The complex V inhibitor oligomycin reduced OCR of the mature adipocytes to 63% of baseline rates, suggesting that 37% of cellular oxygen consumption was coupled to ATP synthesis. The combination of complex I and II inhibitors rotenone and antimycin reduced OCR to 16% of baseline, indicating that mitochondrial fraction accounted for 84% of total cellular respiration. Bioenergetic parameters of naïve and TNF‐α exposed preadipocytes and adipocytes responded differently to certain substrates (glucose, pyruvate, lactate, and palmitate), PPARα/γ activators, and β‐adrenergic agonists, highlighting intrinsic differences in maintaining energy homeostasis during adipocyte differentiation and possible molecular targets for nutritional preemption of metabolic disorders.
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