Using SRM-MS to quantify nuclear protein abundance differences between adipose tissue depots of insulin-resistant mice[S]
| Autor: | Wen-Jun Shen, Robert Ahrends, Fredric B. Kraemer, Olivia Wu, Kyle M. Kovary, Brian J. Feldman, Mary N. Teruel, Asuka Ota, Marco Costa |
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| Jazyk: | angličtina |
| Rok vydání: | 2015 |
| Předmět: |
quantitative proteomics
Male medicine.medical_specialty Biochemistry & Molecular Biology obesity Stromal cell adipocytes Adipose Tissue White Adipose tissue Mice Obese White White adipose tissue QD415-436 Biology Medical Biochemistry and Metabolomics Inbred C57BL Diet High-Fat Biochemistry Obese Mass Spectrometry Cell Line chemistry.chemical_compound Mice Endocrinology Insulin resistance Adipocyte Internal medicine insulin resistance medicine Methods Animals Nuclear protein Nuclear Proteins Cell Biology medicine.disease Diet Mice Inbred C57BL High-Fat chemistry Adipose Tissue Adipogenesis Cell culture peroxisome proliferator-activated receptors selected reaction monitoring mass spectrometry Biochemistry and Cell Biology |
| Zdroj: | Journal of Lipid Research Journal of Lipid Research, Vol 56, Iss 5, Pp 1068-1078 (2015) Journal of lipid research, vol 56, iss 5 |
| ISSN: | 1539-7262 0022-2275 |
| Popis: | Insulin resistance (IR) underlies metabolic disease. Visceral, but not subcutaneous, white adipose tissue (WAT) has been linked to the development of IR, potentially due to differences in regulatory protein abundance. Here we investigate how protein levels are changed in IR in different WAT depots by developing a targeted proteomics approach to quantitatively compare the abundance of 42 nuclear proteins in subcutaneous and visceral WAT from a commonly used insulin-resistant mouse model, Lepr(db/db), and from C57BL/6J control mice. The most differentially expressed proteins were important in adipogenesis, as confirmed by siRNA-mediated depletion experiments, suggesting a defect in adipogenesis in visceral, but not subcutaneous, insulin-resistant WAT. Furthermore, differentiation of visceral, but not subcutaneous, insulin-resistant stromal vascular cells (SVCs) was impaired. In an in vitro approach to understand the cause of this impaired differentiation, we compared insulin-resistant visceral SVCs to preadipocyte cell culture models made insulin resistant by different stimuli. The insulin-resistant visceral SVC protein abundance profile correlated most with preadipocyte cell culture cells treated with both palmitate and TNFα. Together, our study introduces a method to simultaneously measure and quantitatively compare nuclear protein expression patterns in primary adipose tissue and adipocyte cell cultures, which we show can reveal relationships between differentiation and disease states of different adipocyte tissue types. |
| Databáze: | OpenAIRE |
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