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Impaired insulin secretion and β-cell loss in tissue-specific knockout mice with mitochondrial diabetes

Nature Genetics volume 26, pages 336–340 (2000)Cite this article

Abstract

Mitochondrial dysfunction is an important contributor to human pathology1,2,3,4 and it is estimated that mutations of mitochondrial DNA (mtDNA) cause approximately 0.5–1% of all types of diabetes mellitus5,6. We have generated a mouse model for mitochondrial diabetes by tissue-specific disruption of the nuclear gene encoding mitochondrial transcription factor A (Tfam, previously mtTFA; ref. 7) in pancreatic β-cells. This transcriptional activator is imported to mitochondria, where it is essential for mtDNA expression and maintenance8,9. The Tfam-mutant mice developed diabetes from the age of approximately 5 weeks and displayed severe mtDNA depletion, deficient oxidative phosphorylation and abnormal appearing mitochondria in islets at the ages of 7–9 weeks. We performed physiological studies of β-cell stimulus–secretion coupling in islets isolated from 7–9-week-old mutant mice and found reduced hyperpolarization of the mitochondrial membrane potential, impaired Ca2+-signalling and lowered insulin release in response to glucose stimulation. We observed reduced β-cell mass in older mutants. Our findings identify two phases in the pathogenesis of mitochondrial diabetes; mutant β-cells initially display reduced stimulus–secretion coupling, later followed by β-cell loss. This animal model reproduces the β-cell pathology of human mitochondrial diabetes and provides genetic evidence for a critical role of the respiratory chain in insulin secretion.

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Figure 1: Genotyping of different tissues in mutant mice (TfamloxP/TfamloxP, +/RIP-cre) and morphological studies of pancreas in mutant mice and controls (TfamloxP/TfamloxP).
Figure 2: Chemical blood analyses and morphometrical analyses in mutant mice (TfamloxP/TfamloxP, +/RIP-cre; black lines and bars) and littermate controls (TfamloxP/TfamloxP; grey lines and bars).
Figure 3: Immunohistochemical analyses to detect insulin, glucagon and somatostatin producing cells in pancreatic sections from mutant mice (TfamloxP/TfamloxP, +/RIP-cre) and littermate controls (TfamloxP/TfamloxP) at the ages of 7 and 39 weeks.
Figure 4: Measurements of mitochondrial membrane potential and [Ca2+]i in response to glucose in islets of mutant mice (TfamloxP/TfamloxP, +/RIP-cre) and controls (TfamloxP/TfamloxP) at 7–8 weeks.
Figure 5: Measurements of insulin release and insulin content in islets from mutant mice (TfamloxP/TfamloxP, +/RIP-cre; black line and bar) and controls (TfamloxP/TfamloxP; grey line and bar) at the ages of 7–8 weeks.

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Acknowledgements

N.-G.L. is supported by grants from the Swedish Medical Research Council, Swedish Heart and Lung Foundation, Petrus och Augusta Hedlunds Stiftelse, Emil och Vera Cornells Stiftelse, Funds of Karolinska Institutet and Novo Nordisk Pharma AB. C.G. is supported by a grant from the Swedish Diabetes Association, The Swedish Society of Medicine and Knut and Alice Wallenberg Stiftelse. J.S. is supported by a stipend from the Roche Research Foundation. A.O. is supported by a grant from the Swedish Medical Research Council. P.-O.B. is supported by grants from the Swedish Medical Research Council, the Swedish Diabetes Association, the Nordic Insulin Foundation Committee, Funds of Karolinska Institutet, Berth von Kantzows Foundation, Juvenile Diabetes Foundation International, the Swedish Society for Medical Research and the Novo Nordisk Foundation.

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Authors and Affiliations

  1. Department of Molecular Medicine, Karolinska Institutet, Karolinska Hospital, Stockholm, Sweden

    José P. Silva, Caroline Graff & Nils-Göran Larsson

  2. Department of Molecular Medicine, The Rolf Luft Center for Diabetes Research, Karolinska Institutet, Karolinska Hospital, Stockholm, Sweden

    Martin Köhler & Per-Olof Berggren

  3. Department of Pathology, Sahlgrenska University Hospital, Göteborg, Sweden

    Anders Oldfors

  4. Department of Molecular Physiology and Biophysics, Vanderbilt University School of Medicine, Nashville, Tennessee, USA

    Mark A. Magnuson

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  1. José P. Silva
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  2. Martin Köhler
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  3. Caroline Graff
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  6. Per-Olof Berggren
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  7. Nils-Göran Larsson
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Correspondence to Nils-Göran Larsson.

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Silva, J., Köhler, M., Graff, C. et al. Impaired insulin secretion and β-cell loss in tissue-specific knockout mice with mitochondrial diabetes. Nat Genet 26, 336–340 (2000). https://doi.org/10.1038/81649

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