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Prion Infection of Muscle Cells In Vitro

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Journal of Virology, May 2007, p. 4615-4624, Vol. 81, No. 9
0022-538X/07/$08.00+0 doi:10.1128/JVI.02628-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Prion Infection of Muscle Cells In Vitro
Wendy M. Dlakic, Eric Grigg, and Richard A. Bessen*
Department of Veterinary Molecular Biology, Montana State University, Bozeman, Montana 59717

Received 28 November 2006/ Accepted 9 February 2007

The prion agent has been detected in skeletal muscle of humans and animals with prion diseases. Here we report scrapie infection of murine C2C12 myoblasts and myotubes in vitro following coculture with a scrapie-infected murine neuroblastoma (N2A) cell line but not following incubation with a scrapie-infected nonneuronal cell line or a scrapie brain homogenate. Terminal differentiation of scrapie-infected C2C12 myoblasts into myotubes resulted in an increase in the expression of the disease-specific prion protein, PrPSc. The amount of scrapie infectivity or PrPSc in C2C12 myotubes was comparable to the levels found in scrapie-infected N2A cells, indicating that a high level of infection was established in muscle cells. Subclones of scrapie-infected C2C12 cells produced high levels of PrPSc in myotubes, and the C-terminal C2 polypeptide fragment of PrPSc was found based on deglycosylation and PrPSc-specific immunoprecipitation of cell lysates. This is the first report of a stable prion infection in muscle cells in vitro and of a long-term prion infection in a nondividing, differentiated peripheral cell type in culture. These in vitro studies also suggest that in vivo prion infection of skeletal muscle requires contact with prion-infected neurons or, possibly, nerve terminals.



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* Corresponding author. Mailing address: Department of Veterinary Molecular Biology, P.O. Box 173610, Montana State University, Bozeman, MT 59717. Phone: (406) 994-1563. Fax: (406) 994-4303. E-mail: [email protected]

Published ahead of print on 21 February 2007.



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Journal of Virology, May 2007, p. 4615-4624, Vol. 81, No. 9
0022-538X/07/$08.00+0 doi:10.1128/JVI.02628-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

http://jvi.asm.org/cgi/content/abstract/81/9/4615?maxtoshow=&HITS=10&hits=10&RESULTFORMAT=&fulltext=prion&searchid=1&FIRSTINDEX=0&volume=81&issue=9&resourcetype=HWCIT


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Published online before print April 9, 2007
Proc. Natl. Acad. Sci. USA, 10.1073/pnas.0608885104

Medical Sciences
Inducible overexpression of wild-type prion protein in the muscles leads to a primary myopathy in transgenic mice
( doxycycline | inducible transgenic mice | muscle disease )

Shenghai Huang , Jingjing Liang , Mengjie Zheng , Xinyi Li , Meiling Wang , Ping Wang , Difernando Vanegas , Di Wu , Bikram Chakraborty , Arthur P. Hays , Ken Chen ¶, Shu G. Chen , Stephanie Booth ||, Mark Cohen **, Pierluigi Gambetti , and Qingzhong Kong
Department of Pathology, Case Western Reserve University, Cleveland, OH 44106; Department of Cell Biology, Cleveland Clinic Foundation, Cleveland, OH 44195; Department of Pathology, Columbia University Medical Center, New York, NY 10032; ¶Department of Developmental and Molecular Biology, Albert Einstein College of Medicine, Bronx, NY 10461; ||Division of Host Genetics and Prion Diseases National Microbiology Laboratory, Winnipeg, MB, Canada R3E 3R2; and **Institute of Pathology, Case Medical Center, Cleveland, OH 44106


Edited by Stanley B. Prusiner, University of California, San Francisco, CA, and approved March 6, 2007 (received for review October 6, 2006)

The prion protein (PrP) level in muscle has been reported to be elevated in patients with inclusion-body myositis, polymyositis, dermatomyositis, and neurogenic muscle atrophy, but it is not clear whether the elevated PrP accumulation in the muscles is sufficient to cause muscle diseases. We have generated transgenic mice with muscle-specific expression of PrP under extremely tight regulation by doxycycline, and we have demonstrated that doxycycline-induced overexpression of PrP strictly limited to muscles leads to a myopathy characterized by increased variation of myofiber size, centrally located nuclei, and endomysial fibrosis, in the absence of intracytoplasmic inclusions, rimmed vacuoles, or any evidence of a neurogenic disorder. The PrP-induced myopathy correlates with accumulation of an N-terminal truncated PrP fragment in the muscle, and the muscular PrP displayed consistent mild resistance to protease digestion. Our findings indicate that overexpression of wild-type PrP in skeletal muscles is sufficient to cause a primary myopathy with no signs of peripheral neuropathy, possibly due to accumulation of a cytotoxic truncated form of PrP and/or PrP aggregation.



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Author contributions: Q.K. designed research; S.H., J.L., M.Z., X.L., M.W., P.W., D.V., D.W., B.C., A.P.H., S.G.C., P.G., and Q.K. performed research; K.C. and M.C. contributed new reagents/analytic tools; S.H., A.P.H., S.B., M.C., P.G., and Q.K. analyzed data; and P.G. and Q.K. wrote the paper.

The authors declare no conflict of interest.

To whom correspondence should be addressed.

Qingzhong Kong, E-mail: [email protected]

www.pnas.org/cgi/doi/10.1073/pnas.0608885104



http://www.pnas.org/cgi/content/abstract/0608885104v1?etoc





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