. 2015 Dec;64(12):1694-703.

doi: 10.1016/j.metabol.2015.09.008. Epub 2015 Sep 12.

Hepatic proteomic analysis revealed altered metabolic pathways in insulin resistant Akt1(+/-)/Akt2(-/-) mice

Brian A Pedersen¹, Weiwen Wang², Jared F Taylor¹, Omar S Khattab³, Yu-Han Chen⁴, Robert A Edwards⁵, Puya G Yazdi¹, Ping H Wang⁶

Affiliations

¹ UC Irvine Diabetes Center, University of California at Irvine, Irvine, CA 92697, USA; Sue and Bill Gross Stem Cell Research Center, University of California at Irvine, Irvine, CA 92697, USA; Department of Medicine, University of California at Irvine, Irvine, CA 92697, USA.
² UC Irvine Diabetes Center, University of California at Irvine, Irvine, CA 92697, USA; Sue and Bill Gross Stem Cell Research Center, University of California at Irvine, Irvine, CA 92697, USA; Department of Medicine, University of California at Irvine, Irvine, CA 92697, USA; Bascom Palmer Eye Institute, Department of Ophthalmology, University of Miami School of Medicine, Miami, FL, 33136.
³ UC Irvine Diabetes Center, University of California at Irvine, Irvine, CA 92697, USA; Sue and Bill Gross Stem Cell Research Center, University of California at Irvine, Irvine, CA 92697, USA.
⁴ UC Irvine Diabetes Center, University of California at Irvine, Irvine, CA 92697, USA; Sue and Bill Gross Stem Cell Research Center, University of California at Irvine, Irvine, CA 92697, USA; Department of Physiology & Biophysics, University of California at Irvine, Irvine, CA 92697, USA.
⁵ Department of Pathology, University of California at Irvine, Irvine, CA 92697, USA.
⁶ UC Irvine Diabetes Center, University of California at Irvine, Irvine, CA 92697, USA; Sue and Bill Gross Stem Cell Research Center, University of California at Irvine, Irvine, CA 92697, USA; Department of Medicine, University of California at Irvine, Irvine, CA 92697, USA; Department of Biological Chemistry, University of California at Irvine, Irvine, CA 92697, USA; Department of Physiology & Biophysics, University of California at Irvine, Irvine, CA 92697, USA. Electronic address: phwang@uci.edu.

PMID: 26455965
PMCID: PMC4641788
DOI: 10.1016/j.metabol.2015.09.008

Hepatic proteomic analysis revealed altered metabolic pathways in insulin resistant Akt1(+/-)/Akt2(-/-) mice

Brian A Pedersen et al. Metabolism. 2015 Dec.

. 2015 Dec;64(12):1694-703.

doi: 10.1016/j.metabol.2015.09.008. Epub 2015 Sep 12.

Authors

Brian A Pedersen¹, Weiwen Wang², Jared F Taylor¹, Omar S Khattab³, Yu-Han Chen⁴, Robert A Edwards⁵, Puya G Yazdi¹, Ping H Wang⁶

Affiliations

¹ UC Irvine Diabetes Center, University of California at Irvine, Irvine, CA 92697, USA; Sue and Bill Gross Stem Cell Research Center, University of California at Irvine, Irvine, CA 92697, USA; Department of Medicine, University of California at Irvine, Irvine, CA 92697, USA.
² UC Irvine Diabetes Center, University of California at Irvine, Irvine, CA 92697, USA; Sue and Bill Gross Stem Cell Research Center, University of California at Irvine, Irvine, CA 92697, USA; Department of Medicine, University of California at Irvine, Irvine, CA 92697, USA; Bascom Palmer Eye Institute, Department of Ophthalmology, University of Miami School of Medicine, Miami, FL, 33136.
³ UC Irvine Diabetes Center, University of California at Irvine, Irvine, CA 92697, USA; Sue and Bill Gross Stem Cell Research Center, University of California at Irvine, Irvine, CA 92697, USA.
⁴ UC Irvine Diabetes Center, University of California at Irvine, Irvine, CA 92697, USA; Sue and Bill Gross Stem Cell Research Center, University of California at Irvine, Irvine, CA 92697, USA; Department of Physiology & Biophysics, University of California at Irvine, Irvine, CA 92697, USA.
⁵ Department of Pathology, University of California at Irvine, Irvine, CA 92697, USA.
⁶ UC Irvine Diabetes Center, University of California at Irvine, Irvine, CA 92697, USA; Sue and Bill Gross Stem Cell Research Center, University of California at Irvine, Irvine, CA 92697, USA; Department of Medicine, University of California at Irvine, Irvine, CA 92697, USA; Department of Biological Chemistry, University of California at Irvine, Irvine, CA 92697, USA; Department of Physiology & Biophysics, University of California at Irvine, Irvine, CA 92697, USA. Electronic address: phwang@uci.edu.

PMID: 26455965
PMCID: PMC4641788
DOI: 10.1016/j.metabol.2015.09.008

Abstract

Objective: The aim of this study was to identify liver proteome changes in a mouse model of severe insulin resistance and markedly decreased leptin levels.

Methods: Two-dimensional differential gel electrophoresis was utilized to identify liver proteome changes in AKT1(+/-)/AKT2(-/-) mice. Proteins with altered levels were identified with tandem mass spectrometry. Ingenuity Pathway Analysis was performed for the interpretation of the biological significance of the observed proteomic changes.

Results: 11 proteins were identified from 2 biological replicates to be differentially expressed by a ratio of at least 1.3 between age-matched insulin resistant (Akt1(+/-)/Akt2(-/-)) and wild type mice. Albumin and mitochondrial ornithine aminotransferase were detected from multiple spots, which suggest post-translational modifications. Enzymes of the urea cycle were common members of top regulated pathways.

Conclusion: Our results help to unveil the regulation of the liver proteome underlying altered metabolism in an animal model of severe insulin resistance.

Keywords: AKT; Insulin resistance; Liver; Mouse; Proteomics.

PubMed Disclaimer

Conflict of interest statement

Conflict of Interest: The authors have no relevant conflicts of interest to disclose.

Figures

**Figure 1. Analyses of body weights, random serum glucose levels, OGTT and liver histology**
(A) Adult body weights of wild type and Akt1^+/-/Akt2^-/- mutant mice. (B) Random blood glucose levels of wild type and Akt1^+/-/Akt2^-/- mutant mice. (C) Blood glucose levels during OGTT of 20-week- old of wild type and Akt1^+/-/Akt2^-/- mutant mice. (D) Plasma insulin levels during OGTT of 20-week- old of wild type and Akt1^+/-/Akt2^-/- mutant mice. (E) Representative photomicrographs (×200) of liver sections stained with H&E from 20-week-old wild type and Akt1^+/-/Akt2^-/- mutant mice. All results are mean +/- SD. The markings indicate statistical significance in comparison to wild type mice at the same time point (*p<0.001, ^‡p<0.005).

**Figure 2. Representative 2D-DIGE gel of liver proteins**
The gel depicts proteins with an pI 3-7 further separated by molecular weight in a 12.5% polyacrylamide gel. Statistically significant, differentially expressed proteins from both replicates are circled on the above gel. Red circles indicate increased and blue circles indicate decreased normalized protein levels in AKT1^+/-/AKT2^-/- mice in comparison to wild type mice. Information on their identities and differential expression is in Table 1.

**Figure 3. Classification of identified proteins and biological pathway analysis**
(A) Cellular component localization of the 11 differentially expressed proteins identified as per UniProt. For each cellular component, the number of proteins in each class is given. (B) Enriched biological process networks by IPA software for the 11 differentially expressed proteins. The –log(p-value) for the enriched networks is as is per the height of the bars along the y-axis on the left. Upregulated pathways are colored yellow and downregulated pathways are colored blue. The ratio of the number of enriched proteins from our dataset in comparison to the total number of proteins within a pathway is indicated by the orange line and as per the y-axis on the right.

See this image and copyright information in PMC

References

1. Araki E, et al. Alternative pathway of insulin signalling in mice with targeted disruption of the IRS-1 gene. Nature. 1994;372(6502):186–90. - PubMed
1. Withers DJ, et al. Disruption of IRS-2 causes type 2 diabetes in mice. Nature. 1998;391(6670):900–4. - PubMed
1. Liu SC, et al. Insulin receptor substrate 3 is not essential for growth or glucose homeostasis. J Biol Chem. 1999;274(25):18093–9. - PubMed
1. Fantin VR, et al. Mice lacking insulin receptor substrate 4 exhibit mild defects in growth, reproduction, and glucose homeostasis. Am J Physiol Endocrinol Metab. 2000;278(1):E127–33. - PubMed
1. Cho H, et al. Insulin resistance and a diabetes mellitus-like syndrome in mice lacking the protein kinase Akt2 (PKB beta) Science. 2001;292(5522):1728–31. - PubMed

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Hepatic proteomic analysis revealed altered metabolic pathways in insulin resistant Akt1(+/-)/Akt2(-/-) mice

Affiliations

Hepatic proteomic analysis revealed altered metabolic pathways in insulin resistant Akt1(+/-)/Akt2(-/-) mice

Authors

Affiliations

Abstract

Conflict of interest statement

Figures

References

Publication types

MeSH terms

Substances

Grants and funding

LinkOut - more resources

Full Text Sources

Other Literature Sources

Miscellaneous