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. 2004 Nov 23;101(47):16525-30.
doi: 10.1073/pnas.0407574101. Epub 2004 Nov 16.

Distinct pathways of insulin-regulated versus diabetes-regulated gene expression: an in vivo analysis in MIRKO mice

Vijay K Yechoor  1 , Mary-Elizabeth Patti, Kohjiro Ueki, Palle G Laustsen, Robert Saccone, Ravi Rauniyar, C Ronald Kahn
Affiliations

Distinct pathways of insulin-regulated versus diabetes-regulated gene expression: an in vivo analysis in MIRKO mice

Vijay K Yechoor et al. Proc Natl Acad Sci U S A. .

Abstract

Diabetes mellitus is a complex metabolic disorder accompanied by alterations in cellular physiology, metabolism, and gene expression. These alterations can be primary (due to loss of direct insulin action) or secondary (due to the metabolic perturbations associated with the disease). To dissect and quantitate these two separate effects, we compared the skeletal muscle gene-expression profiles of muscle insulin receptor knockout (MIRKO) mice and their Lox controls in the basal, streptozotocin-induced diabetic, and insulin-treated diabetic states. Pure deficiency of insulin action as present in the MIRKO mouse results in regulation of 130 genes, with down-regulation of NSF (N-ethylmaleimide-sensitive fusion protein) and VAMP-2 (vesicle-associated membrane protein 2), stearoyl CoA desaturase 1, and cAMP-specific phosphodiesterase 4B, as well as up-regulation of some signaling-related genes, such as Akt2, and the fatty-acid transporter CD36. In diabetes, additional transcriptional mechanisms are activated, resulting in alterations in expression of approximately 500 genes, including a highly coordinated down-regulation of genes of the mitochondrial electron-transport chain and one of the mammalian homologues of the histone deacetylase Sir2, which has been implicated in the link between nutrition and longevity. These distinct pathways of direct and indirect regulation of gene expression provide insights into the complex mechanisms of transcriptional control in diabetes and areas of potential therapeutic targeting.

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Figures

Fig. 1.
Fig. 1.
Experimental design. (a) MIRKO mice and their Lox control littermates were treated with either STZ or citrate buffer. The diabetic (blood sugar, >400 mg/dl) mice were either followed or treated with insulin (blood sugar, <200 mg/dl) (STZ–insulin group). (b) Genes that are altered significantly in expression in the MIRKO, Lox-STZ, and MIRKO-STZ groups are shown in a Venn diagram. See Results for a detailed explanation.
Fig. 2.
Fig. 2.
Insulin-regulated versus diabetes-regulated gene expression. (a) Comparison of gene expression in Lox-STZ diabetic and MIRKO mice. The log of the ratios of the expression (experimental group/control) of genes that are changed significantly in either MIRKO or the Lox-STZ when compared with the Lox control are plotted on a log scale (every 0.3 units on the scale equals a 2-fold change). This comparison separated the genes into four quadrants, each reflecting either a concordant or discordant regulation of the genes by the loss of insulin-receptor-mediated signaling and the diabetic state. The genes labeled A and D, for example, were altered in diabetes, but they were not altered by a pure loss of insulin action in the MIRKO mouse; in contrast, the genes labeled B and C were altered in the MIRKO mouse but not in STZ diabetes. (b) The log of the ratios of the expression (experimental group/control) of genes that are changed significantly in either the MIRKO or the MIRKO-STZ when compared with the Lox control. The diagonal black line indicates the line of unity.
Fig. 3.
Fig. 3.
The “loss-of-insulin effect” and the calculated “diabetes effect” are shown for representative genes. The loss-of-insulin effect was calculated from the percentage of change in expression in the MIRKO as compared with the Lox controls. The diabetes effect was calculated as the difference between the percentage of change in the MIRKO-STZ and MIRKO when compared with the Lox controls (Table 5, which is published as supporting information on the PNAS web site).
Fig. 4.
Fig. 4.
Contrasting patterns of diabetes- and insulin-regulated genes. (a) The ratios of the expressions (experimental/lox control) of all of the genes of the electron-transport chain that were changed significantly in the diabetic groups are shown. All of these genes are changed significantly in the diabetic groups (Lox-STZ and MIRKO-STZ) but not in the MIRKO group. With insulin treatment, all of these genes corrected toward the Lox control by >50% in the Lox-STZ-INS but not in the MIRKO-STZ-INS group. The indicated genes are subunits of the electron-transport chain complexes I–V (C-I–C-V). (b) The ratios of the expression (experimental/lox-control) of the genes for carnitine palmitoyl CoA transferase1 (CPT1), δ3-δ2 enoyl CoA hydratase, cAMP-dependent protein kinase, and ubiquitin-specific protease 2 are shown. All of these genes are changed significantly in the diabetic groups (Lox-STZ and MIRKO-STZ) but not in the MIRKO group. With insulin treatment, all of these genes corrected toward the control by >50% in the Lox-STZ-INS but not in the MIRKO-STZ-INS group.
Fig. 5.
Fig. 5.
Changes in Sirtuin3 and sir2 with diabetes. (a) The mean transcript levels of Sirtuin3 in skeletal muscle in the various metabolic groups, as detected by microarray analysis, are shown as a percentage of the level in the control group. (b) The bands for sir2 in the nuclear (N) and cytosolic (C) fractions from the hind-limb muscles of wild-type control and STZ-induced diabetic mice are shown on immunoblots (for detailed methods, see Supporting Materials and Methods). (c) The mean intensity of the nuclear and cytosolic fraction sir2 bands on immunoblotting from two control and two diabetic mice are shown. The total is the sum of the respective nuclear and cytosolic fractions. The levels are represented as a percentage of the mean total level in the control group.
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References

    1. Skyler, J. S. & Oddo, C. (2002) Diabetes Metab Res. Rev. 18 Suppl 3, S21–S26. - PubMed
    1. Kahn, C. R. (1994) Diabetes 43, 1066–1084. - PubMed
    1. Yechoor, V. K., Patti, M. E., Saccone, R. & Kahn, C. R. (2002) Proc. Natl. Acad. Sci. USA 99, 10587–10592. - PMC - PubMed
    1. Sreekumar, R., Halvatsiotis, P., Schimke, J. C. & Nair, K. S. (2002) Diabetes 51, 1913–1920. - PubMed
    1. O'Brien, R. M. & Granner, D. K. (1996) Physiol. Rev. 76, 1109–1161. - PubMed

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