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Using SRM-MS to quantify nuclear protein abundance differences between adipose tissue depots of insulin-resistant mice

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 b...

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Autores principales: Ota, Asuka, Kovary, Kyle M., Wu, Olivia H., Ahrends, Robert, Shen, Wen-Jun, Costa, Maria J., Feldman, Brian J., Kraemer, Fredric B., Teruel, Mary N.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The American Society for Biochemistry and Molecular Biology 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4409283/
https://www.ncbi.nlm.nih.gov/pubmed/25840986
http://dx.doi.org/10.1194/jlr.D056317
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author Ota, Asuka
Kovary, Kyle M.
Wu, Olivia H.
Ahrends, Robert
Shen, Wen-Jun
Costa, Maria J.
Feldman, Brian J.
Kraemer, Fredric B.
Teruel, Mary N.
author_facet Ota, Asuka
Kovary, Kyle M.
Wu, Olivia H.
Ahrends, Robert
Shen, Wen-Jun
Costa, Maria J.
Feldman, Brian J.
Kraemer, Fredric B.
Teruel, Mary N.
author_sort Ota, Asuka
collection PubMed
description 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.
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spelling pubmed-44092832015-05-07 Using SRM-MS to quantify nuclear protein abundance differences between adipose tissue depots of insulin-resistant mice Ota, Asuka Kovary, Kyle M. Wu, Olivia H. Ahrends, Robert Shen, Wen-Jun Costa, Maria J. Feldman, Brian J. Kraemer, Fredric B. Teruel, Mary N. J Lipid Res Methods 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. The American Society for Biochemistry and Molecular Biology 2015-05 /pmc/articles/PMC4409283/ /pubmed/25840986 http://dx.doi.org/10.1194/jlr.D056317 Text en http://creativecommons.org/licenses/by/3.0/ Author’s Choice—Final version full access. Creative Commons Attribution Unported License (http://creativecommons.org/licenses/by/3.0/) applies to Author Choice Articles
spellingShingle Methods
Ota, Asuka
Kovary, Kyle M.
Wu, Olivia H.
Ahrends, Robert
Shen, Wen-Jun
Costa, Maria J.
Feldman, Brian J.
Kraemer, Fredric B.
Teruel, Mary N.
Using SRM-MS to quantify nuclear protein abundance differences between adipose tissue depots of insulin-resistant mice
title Using SRM-MS to quantify nuclear protein abundance differences between adipose tissue depots of insulin-resistant mice
title_full Using SRM-MS to quantify nuclear protein abundance differences between adipose tissue depots of insulin-resistant mice
title_fullStr Using SRM-MS to quantify nuclear protein abundance differences between adipose tissue depots of insulin-resistant mice
title_full_unstemmed Using SRM-MS to quantify nuclear protein abundance differences between adipose tissue depots of insulin-resistant mice
title_short Using SRM-MS to quantify nuclear protein abundance differences between adipose tissue depots of insulin-resistant mice
title_sort using srm-ms to quantify nuclear protein abundance differences between adipose tissue depots of insulin-resistant mice
topic Methods
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4409283/
https://www.ncbi.nlm.nih.gov/pubmed/25840986
http://dx.doi.org/10.1194/jlr.D056317
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