Muscle-specific IRS-1 Ser->Ala transgenic mice are protected from fat-induced insulin resistance in skeletal muscle - PubMed (original) (raw)
. 2008 Oct;57(10):2644-51.
doi: 10.2337/db06-0454. Epub 2008 Jul 15.
Susanne Neschen, Stefan Bilz, Saki Sono, Dimitrios Tsirigotis, Richard M Reznick, Irene Moore, Yoshio Nagai, Varman Samuel, David Sebastian, Morris White, William Philbrick, Gerald I Shulman
Affiliations
- PMID: 18633112
- PMCID: PMC2551673
- DOI: 10.2337/db06-0454
Muscle-specific IRS-1 Ser->Ala transgenic mice are protected from fat-induced insulin resistance in skeletal muscle
Katsutaro Morino et al. Diabetes. 2008 Oct.
Abstract
Objective: Insulin resistance in skeletal muscle plays a critical role in the pathogenesis of type 2 diabetes, yet the cellular mechanisms responsible for insulin resistance are poorly understood. In this study, we examine the role of serine phosphorylation of insulin receptor substrate (IRS)-1 in mediating fat-induced insulin resistance in skeletal muscle in vivo.
Research design and methods: To directly assess the role of serine phosphorylation in mediating fat-induced insulin resistance in skeletal muscle, we generated muscle-specific IRS-1 Ser(302), Ser(307), and Ser(612) mutated to alanine (Tg IRS-1 Ser-->Ala) and IRS-1 wild-type (Tg IRS-1 WT) transgenic mice and examined insulin signaling and insulin action in skeletal muscle in vivo.
Results: Tg IRS-1 Ser-->Ala mice were protected from fat-induced insulin resistance, as reflected by lower plasma glucose concentrations during a glucose tolerance test and increased insulin-stimulated muscle glucose uptake during a hyperinsulinemic-euglycemic clamp. In contrast, Tg IRS-1 WT mice exhibited no improvement in glucose tolerance after high-fat feeding. Furthermore, Tg IRS-1 Ser-->Ala mice displayed a significant increase in insulin-stimulated IRS-1-associated phosphatidylinositol 3-kinase activity and Akt phosphorylation in skeletal muscle in vivo compared with WT control littermates.
Conclusions: These data demonstrate that serine phosphorylation of IRS-1 plays an important role in mediating fat-induced insulin resistance in skeletal muscle in vivo.
Figures
FIG. 1.
Generation of Tg IRS-1 Ser→Ala mice. A: Serine phosphorylation of IRS-1 protein in skeletal muscle. Serine phosphorylation of IRS-1 and protein expression of IRS-1 were analyzed by Western blotting. All mice were killed at age 16 weeks. The high-fat diet group was fed for 8 weeks. B: Vector construct of the mutated IRS-1 gene (Ser302, Ser307, and Ser612 to Ala mutant) for Tg IRS-1 Ser→Ala and WT IRS-1 gene for Tg IRS-1 WT. C: Comparison of IRS-1 expression in different insulin-targeting tissues. Protein expression of total IRS-1 and actin were analyzed by Western blotting. D: Comparison of IRS-1 expression between Tg IRS-1 Ser→Ala and Tg IRS-1 WT mice. Each graph was expressed as fold difference to their littermates. GAS, Musculus gastrocnemius; QD, Musclus quadriceps; TA, Musculus tibialis anterior; WAT, white adipose tissue.
FIG. 2.
Tg IRS-1 Ser→Ala, but not Tg IRS-1 WT, mice were protected from high-fat diet–induced glucose intolerance during IPGTT. A: Growth curves. Body weights were measured weekly under control diet and high-fat diet feeding regimens. At age 7–8 weeks, mice were started on a high-fat, safflower oil–based diet (27% safflower oil, 59% fat-derived calories) and maintained for 8 weeks. with solid line, WT control diet; • with solid line, Tg IRS-1 Ser→Ala control diet; with dotted line, WT high-fat diet; • with dotted line, Tg IRS-1 Ser→Ala high-fat diet. B: IPGTT. A total of 1 g/kg of 10% glucose was injected in control diet–fed Tg IRS-1 Ser→Ala mice (• with solid line) and littermate control mice ( with solid line). A total of 1 g/kg of 10% glucose was also injected in high-fat–fed Tg IRS-1 Ser→Ala (• with dotted line), Tg IRS-1 WT (♦ with dotted line), and littermate control ( with dotted line) mice. C: Area under the curve of insulin concentration during IPGTTs. Results are expressed as means ± SE (WT, n = 11; Tg IRS-1 Ser→Ala, n = 16; Tg IRS-1 WT, n = 8). *P < 0.05 vs. WT high-fat diet.
FIG. 3.
Tg IRS-1 Ser→Ala mice were protected from high-fat diet–induced muscle insulin resistance. A: After 8 weeks of high-fat feeding (16 ± 1 weeks), age-matched mice were food deprived for 16 h and hyperinsulinemic-euglycemic clamp experiments (2.5 mU · kg−1 · min−1) were performed using Tg IRS-1 Ser→Ala mice and their littermates. Plasma glucose concentration was maintained by an intravenous 20% glucose infusion and glucose infusion rates (GINF) at steady state were assessed. B: Muscle-specific glucose uptake was analyzed by enrichment of [14C]-2-deoxy-glucose in Musculus gastrocnemius (GAS) (left) and Musclus quadriceps (QD) (right). Results are expressed as means ± SE (WT, n = 12; Tg IRS-1 Ser→Ala, n = 16). C: Insulin-stimulated Akt phosphorylation (Ser473) in GAS was analyzed 15 min after intraperitoneal injection of 0.6 units/kg insulin in control diet–fed mice (WT, n = 6) and high-fat diet–fed mice (WT, n = 16; Tg IRS-1 Ser→Ala, n = 15; Tg IRS-1 WT, n = 3). D: IRS-1–associated PI3-kinase activity in GAS of high-fat–fed mice (WT, n = 12; Tg IRS-1 Ser→Ala, n = 8). E: IRS-1 tyrosine phosphorylation (left) and IRS-1–associated p85 subunit of PI3-kinase (right) were analyzed 15 min after intraperitoneal injection of 0.6 units/kg insulin by immunoprecipitation. Results are expressed as means relative to insulin-stimulated samples of WT high-fat fed mice ± SE.
FIG. 4.
Tg IRS-1 Ser→Ala mice were protected from age-related glucose intolerance. Plasma glucose (A) and insulin (B) concentrations in 1-year-old Tg IRS-1 Ser→Ala transgenic and WT littermate mice following an IPGTT (1 g/kg) (WT, n = 12; Tg IRS-1 Ser→Ala, n = 6). A: WT elder; ♦, Tg IRS-1 Ser→Ala elder. Results are expressed as means ± SE. C: Generation of Tg IRS-1 Ser→Ala/ob/ob. Growth curves: body weights were measured weekly under control diet until age 16 weeks (• with dotted line) and compared with WT/ob/ob ( with dotted line), Tg IRS-1 Ser→Ala/+/+ (• with solid line), and WT/+/+ ( with solid line); n = 7–10. *P < 0.05 vs. WT littermate mice.
Similar articles
- Short-term in vivo inhibition of insulin receptor substrate-1 expression leads to insulin resistance, hyperinsulinemia, and increased adiposity.
Araújo EP, De Souza CT, Gasparetti AL, Ueno M, Boschero AC, Saad MJ, Velloso LA. Araújo EP, et al. Endocrinology. 2005 Mar;146(3):1428-37. doi: 10.1210/en.2004-0778. Epub 2004 Nov 18. Endocrinology. 2005. PMID: 15550510 - Inducible nitric oxide synthase deficiency ameliorates skeletal muscle insulin resistance but does not alter unexpected lower blood glucose levels after burn injury in C57BL/6 mice.
Sugita M, Sugita H, Kim M, Mao J, Yasuda Y, Habiro M, Shinozaki S, Yasuhara S, Shimizu N, Martyn JA, Kaneki M. Sugita M, et al. Metabolism. 2012 Jan;61(1):127-36. doi: 10.1016/j.metabol.2011.06.001. Epub 2011 Aug 3. Metabolism. 2012. PMID: 21816442 Free PMC article. - Caffeine modulates phosphorylation of insulin receptor substrate-1 and impairs insulin signal transduction in rat skeletal muscle.
Egawa T, Tsuda S, Ma X, Hamada T, Hayashi T. Egawa T, et al. J Appl Physiol (1985). 2011 Dec;111(6):1629-36. doi: 10.1152/japplphysiol.00249.2011. Epub 2011 Sep 22. J Appl Physiol (1985). 2011. PMID: 21940847 - p38 mitogen-activated protein kinase-dependent transactivation of ErbB receptor family: a novel common mechanism for stress-induced IRS-1 serine phosphorylation and insulin resistance.
Hemi R, Yochananov Y, Barhod E, Kasher-Meron M, Karasik A, Tirosh A, Kanety H. Hemi R, et al. Diabetes. 2011 Apr;60(4):1134-45. doi: 10.2337/db09-1323. Epub 2011 Mar 8. Diabetes. 2011. PMID: 21386087 Free PMC article. - The role of endothelial insulin signaling in the regulation of glucose metabolism.
Kubota T, Kubota N, Kadowaki T. Kubota T, et al. Rev Endocr Metab Disord. 2013 Jun;14(2):207-16. doi: 10.1007/s11154-013-9242-z. Rev Endocr Metab Disord. 2013. PMID: 23589150 Review.
Cited by
- An estrogen receptor α-derived peptide improves glucose homeostasis during obesity.
Yang W, Jiang W, Liao W, Yan H, Ai W, Pan Q, Brashear WA, Xu Y, He L, Guo S. Yang W, et al. Nat Commun. 2024 Apr 22;15(1):3410. doi: 10.1038/s41467-024-47687-6. Nat Commun. 2024. PMID: 38649684 Free PMC article. - The serine phosphorylations in the IRS-1 PIR domain abrogate IRS-1 and IR interaction.
Woo JR, Bae SH, Wales TE, Engen JR, Lee J, Jang H, Park S. Woo JR, et al. Proc Natl Acad Sci U S A. 2024 Apr 23;121(17):e2401716121. doi: 10.1073/pnas.2401716121. Epub 2024 Apr 16. Proc Natl Acad Sci U S A. 2024. PMID: 38625937 - _O_-linked _N_-acetylglucosamine modification is essential for physiological adipose expansion induced by high-fat feeding.
Nakamoto A, Ohashi N, Sugawara L, Morino K, Ida S, Perry RJ, Sakuma I, Yanagimachi T, Fujita Y, Ugi S, Kume S, Shulman GI, Maegawa H. Nakamoto A, et al. Am J Physiol Endocrinol Metab. 2023 Jul 1;325(1):E46-E61. doi: 10.1152/ajpendo.00263.2022. Epub 2023 May 24. Am J Physiol Endocrinol Metab. 2023. PMID: 37224467 Free PMC article. - Insights into the development of insulin resistance: Unraveling the interaction of physical inactivity, lipid metabolism and mitochondrial biology.
Handy RM, Holloway GP. Handy RM, et al. Front Physiol. 2023 Apr 20;14:1151389. doi: 10.3389/fphys.2023.1151389. eCollection 2023. Front Physiol. 2023. PMID: 37153211 Free PMC article. Review. - Links between Insulin Resistance and Periodontal Bacteria: Insights on Molecular Players and Therapeutic Potential of Polyphenols.
Thouvenot K, Turpin T, Taïlé J, Clément K, Meilhac O, Gonthier MP. Thouvenot K, et al. Biomolecules. 2022 Feb 28;12(3):378. doi: 10.3390/biom12030378. Biomolecules. 2022. PMID: 35327570 Free PMC article. Review.
References
- Shulman GI, Rothman DL, Jue T, Stein P, DeFronzo RA, Shulman RG: Quantitation of muscle glycogen-synthesis in normal subjects and subjects with non-insulin-dependent diabetes by C-13 nuclear magnetic-resonance spectroscopy. N Engl J Med 322:223–228, 1990 - PubMed
- Yu CL, Chen Y, Cline GW, Zhang DY, Zong HH, Wang YL, Bergeron R, Kim JK, Cushman SW, Cooney GJ, Atcheson B, White MF, Kraegen EW, Shulman GI: Mechanism by which fatty acids inhibit insulin activation of insulin receptor substrate-1 (IRS-1)-associated phosphatidylinositol 3-kinase activity in muscle. J Biol Chem 277:50230–50236, 2002 - PubMed
- Griffin ME, Marcucci MJ, Cline GW, Bell K, Barucci N, Lee D, Goodyear LJ, Kraegen EW, White MF, Shulman GI: Free fatty acid–induced insulin resistance is associated with activation of protein kinase C θ and alterations in the insulin signaling cascade. Diabetes 48:1270–1274, 1999 - PubMed
Publication types
MeSH terms
Substances
Grants and funding
- U24 DK076169/DK/NIDDK NIH HHS/United States
- R01 DK-40936/DK/NIDDK NIH HHS/United States
- HHMI/Howard Hughes Medical Institute/United States
- U24 DK059635/DK/NIDDK NIH HHS/United States
- P30 DK-45735/DK/NIDDK NIH HHS/United States
- R01 DK040936/DK/NIDDK NIH HHS/United States
- P30 DK045735/DK/NIDDK NIH HHS/United States
- U24 DK-76169/DK/NIDDK NIH HHS/United States
LinkOut - more resources
Full Text Sources
Molecular Biology Databases
Miscellaneous