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Neuronal overexpression of insulin receptor substrate 2 leads to increased fat mass, insulin resistance, and glucose intolerance during aging

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Abstract

The insulin receptor substrates (IRS) are adapter proteins mediating insulin's and IGF1's intracellular effects. Recent data suggest that IRS2 in the central nervous system (CNS) is involved in regulating fuel metabolism as well as memory formation. The present study aims to specifically define the role of chronically increased IRS2-mediated signal transduction in the CNS. We generated transgenic mice overexpressing IRS2 specifically in neurons (nIRS2 tg) and analyzed these in respect to energy metabolism, learning, and memory. Western blot (WB) analysis of nIRS2 tg brain lysates revealed increased IRS2 downstream signaling. Histopathological investigation of nIRS2 tg mice proved unaltered brain development and structure. Interestingly, nIRS2 tg mice showed decreased voluntary locomotoric activity during dark phase accompanied with decreased energy expenditure (EE) leading to increased fat mass. Accordingly, nIRS2 tg mice develop insulin resistance and glucose intolerance during aging. Exploratory behavior, motor function as well as food and water intake were unchanged in nIRS2 tg mice. Surprisingly, increased IRS2-mediated signals did not change spatial working memory in the T-maze task. Since FoxO1 is a key mediator of IRS2-transmitted signals, we additionally generated mice expressing a dominant negative mutant of FoxO1 (FoxO1DN) specifically in neurons. This mutant mimics the effect of increased IRS2 signaling on FoxO-mediated transcription. Interestingly, the phenotype observed in nIRS2 tg mice was not present in FoxO1DN mice. Therefore, increased neuronal IRS2 signaling causes decreased locomotoric activity in the presence of unaltered exploratory behavior and motor coordination that might lead to increased fat mass, insulin resistance, and glucose intolerance during aging independent of FoxO1-mediated transcription.

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Acknowledgments

This work was supported by AFI #08813. JZ was supported by a student's grant of the Medical Faculty, University of Cologne. We thank Andre Kleinridders for his kind introduction and troubleshooting in transgenic mouse generation. Also, we thank Thomas Wunderlich for kindly providing the pCAGGS targeting vector. Thanks also to Jens Alber for excellent technical assistance, Prof. Wilhelm Stoffel for providing the T-maze, and Prof. Martina Deckert and Mariana Carstov for advice and help on immunohistochemistry.

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Authors and Affiliations

  1. Center for Endocrinology, Diabetes and Preventive Medicine (CEDP), University of Cologne, Kerpener Str. 62, 50937, Cologne, Germany

    J. Zemva, M. Udelhoven, L. Moll, S. Freude, O. Stöhr, R. B. Drake, W. Krone & M. Schubert

  2. Center for Molecular Medicine Cologne (CMMC), University of Cologne, Robert-Koch-Str. 21, 50931, Cologne, Germany

    J. Zemva, M. Udelhoven, L. Moll, S. Freude, O. Stöhr, R. B. Drake, W. Krone & M. Schubert

  3. Cologne Cluster of Excellence in Cellular Stress Responses in Aging-associated Diseases (CECAD), University of Cologne, Cologne, Germany

    H. S. Brönneke, W. Krone & M. Schubert

  4. Biochemistry and Molecular Biology, Institute for Molecular Research Israel—Canada (IMRIC), School of Medicine of the Hebrew University of Jerusalem, Jerusalem, 91120, Israel

    L. Moll

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  1. J. Zemva
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  2. M. Udelhoven
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  3. L. Moll
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  4. S. Freude
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  5. O. Stöhr
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  6. H. S. Brönneke
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  7. R. B. Drake
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  8. W. Krone
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  9. M. Schubert
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Corresponding author

Correspondence to M. Schubert.

Additional information

J. Zemva and M. Udelhoven contributed equally to this study.

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Suppl. Fig. 1

Phenotype of WT, nIRS2 tg and homozygous (homo) nIRS2 tg mice at the age of 60 weeks. Transgenic mice appear to have increased abdominal fat mass during aging. The phenotype was more pronounced in homozygous compared to heterozygous mice (JPEG 41 kb)

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Suppl. Fig. 2

Exploratory behavior and motor coordination of male nIRS2 tg mice and respective controls. a Open field test. b Elevated O-Maze test. c Rotarod test, accel: latency to fall [s] at constantly increasing rounds per minute (rpm). Data are expressed as means ± SEM, n = 6 per group, * p < 0.05 (Student's t test), age: 40 weeks (JPEG 45 kb)

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Suppl. Fig. 3

Spatial memory and hippocampal function of male nIRS2 tg mice and respective controls. a Enclosed T-maze test. Data represent means ± SEM, n = 6 per group, * p < 0.05 (Student's t test), age: 20 and 100 weeks (JPEG 7 kb)

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Zemva, J., Udelhoven, M., Moll, L. et al. Neuronal overexpression of insulin receptor substrate 2 leads to increased fat mass, insulin resistance, and glucose intolerance during aging. AGE 35, 1881–1897 (2013). https://doi.org/10.1007/s11357-012-9491-x

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