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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...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The American Society for Biochemistry and Molecular Biology
2015
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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. |
format | Online Article Text |
id | pubmed-4409283 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | The American Society for Biochemistry and Molecular Biology |
record_format | MEDLINE/PubMed |
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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