Abstract
Energy consuming, heat-producing beige adipocytes, located in classic white adipose tissue (WAT), hold promise for the treatment of obesity. Few reports have quantitatively assessed the contribution of browned 'WAT' to energy expenditure. There is a need for methods to examine beige-fat thermogenesis, independently of classical brown fat. The aim of this study is to optimize an inducible lentiviral shRNA to conditionally knock-down Ucp1 and assess the effects on 'browned' WAT. Primary adipocytes from mouse inguinal WAT converted into thermogenic adipocytes when stimulated with β-adrenergic agonist and thiazolidinedione. There was increased UCP1 protein and importantly increases in various indicators of mitochondrial bioenergetics. Next, we determined optimal transfection conditions for the UCP1-shRNA lentiviral system and subsequently applied this to 'browned' WAT. UCP1 knockdown decreased the brown/beige-fat gene profile and decreased mitochondrial respiration. In summary, this study optimizes lentiviral UCP1-shRNA technology in vitro. This technique could be applied to inguinal fat depots in vivo. This would allow investigation of contribution of depots to whole-body metabolism to help elucidate the physiological relevance of beige fat.
Highlights
For the first time in human history the global burden of overnutrition outpaces that due to undernutrition
Beige adipocytes from C57BL/6J mice under mitochondrial stress test demonstrate increased oxygen consumption rates in the presence of Rosiglitazone and CL316, 253 (Fig 2A and 2B)
ATP respiration increased by approximately 2.5 fold in both rosiglitazone (p
Summary
For the first time in human history the global burden of overnutrition outpaces that due to undernutrition. Worldwide overweight and obesity accounts for at least 2.8 million deaths each year and 35.8 million disability-adjusted life years [1]. Obesity is a pathological state characterized by accumulation of excess adipose tissue and lipids. It increases the risk of glucose intolerance, hypertension and dyslipidemia. Obesity contributes to cardiovascular disease, type 2 diabetes mellitus and many cancers. Effective treatment regimens are scarce and new therapeutic targets are needed. Given its central role in energy and nutrient homeostasis, many have looked towards adipose tissue for a solution
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