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Analysis of gene expression differences between utrophin/dystrophin-deficient vs mdx skeletal muscles reveals a specific upregulation of slow muscle genes in limb muscles

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Abstract

Dystrophin deficiency leads to the progressive muscle wasting disease Duchenne muscular dystrophy (DMD). Dystrophin-deficient mdx mice are characterized by skeletal muscle weakness and degeneration but they appear outwardly normal in contrast to DMD patients. Mice lacking both dystrophin and the dystrophin homolog utrophin [double knockout (dko)] have muscle degeneration similar to mdx mice, but they display clinical features similar to DMD patients. Dko limb muscles also lack postsynaptic membrane folding and display fiber-type abnormalities including an abundance of phenotypically oxidative muscle fibers. Extraocular muscles, which are spared in mdx mice, show a significant pathology in dko mice. In this study, microarray analysis was used to characterize gene expression differences between mdx and dko tibialis anterior and extraocular skeletal muscles in an effort to understand the phenotypic differences between these two dystrophic mouse models. Analysis of gene expression differences showed that upregulation of slow muscle genes specifically characterizes dko limb muscle and suggests that upregulation of these genes may directly account for the more severe phenotype of dko mice. To investigate whether any upregulation of slow genes is retained in vitro, independent of postsynaptic membrane abnormalities, we derived mdx and dko primary myogenic cultures and analyzed the expression of Myh7 and Myl2. Real-time reverse transcriptase-polymerase chain reaction analysis demonstrates that transcription of these slow genes is also upregulated in dko vs mdx myotubes. This data suggests that at least part of the fiber-type abnormality is due directly to the combined absence of utrophin and dystrophin and is not an indirect effect of the postsynaptic membrane abnormalities.

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Acknowledgements

This work was supported by the Muscular Dystrophy Association (to JRF), NIH R01 EY12779 (to JDP), and in part, by a Career Award in the Biomedical Sciences from the Burroughs Wellcome Fund (to JRF). PEB was supported by an NIH supplement (AR47034-S). We would like to thank Jaimy Lekan, Katherine L. Gardner, Jonathan Edwards, and Chad Groer for technical assistance and the Biochemistry and Molecular Biology Core, Department of Veterinary Biosciences, OSU for the use of the real-time PCR machine.

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Authors and Affiliations

  1. Department of Molecular and Cellular Biochemistry, College of Medicine, The Ohio State University, Columbus, OH, USA

    Patrick E. Baker, Jessica A. Kearney, Donald E. Kuhn & Jill A. Rafael-Fortney

  2. Department of Neurology, Case Western Reserve University, Cleveland, OH, USA

    Bendi Gong, Anita P. Merriam & John D. Porter

  3. University Hospitals of Cleveland, Cleveland, OH, USA

    Bendi Gong, Anita P. Merriam & John D. Porter

  4. National Institutes of Neurological Disorders and Stroke, 6001 Executive Blvd, NINDS/NSC, Bethesda, MD, 2142, USA

    John D. Porter

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  1. Patrick E. Baker
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  2. Jessica A. Kearney
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Correspondence to Jill A. Rafael-Fortney.

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Baker, P.E., Kearney, J.A., Gong, B. et al. Analysis of gene expression differences between utrophin/dystrophin-deficient vs mdx skeletal muscles reveals a specific upregulation of slow muscle genes in limb muscles. Neurogenetics 7, 81–91 (2006). https://doi.org/10.1007/s10048-006-0031-7

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