Summary
The development of therapies, based upon implantation of normal muscle cell precursors, for the treatment of skeletal muscle diseases such as Duchenne Muscular Dystrophy is in its infancy. Detailed analysis of the genetic and phenotypic contribution made by donor myoblasts to the regenerated muscle is critical. Using non-radioactivein situ hybridization of aY chromosome-specific DNA probe to sections of muscle, we have localized the position of male donor nuclei within female host muscles after myoblast implantation. These results were compared with the distribution of immunocytochemically-localized dystrophin and the expression of donor-specific glucose phosphate isomerase by isoelectric-focussing. We found consistent male-specific nuclear hybridization and a close spatial relationship between the distribution of male donor nuclei and dystrophin-positive muscle fibres within female, dystrophin-negative host muscles. This approach will be useful in the further analysis of myoblast implantation experiments.
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Bishop, C. E., Boursot, P., Baron, B., Bonhomme, P., &Hatat, D. (1985) Most classical Mus musculus laboratory mouse strains carry aMus musculus Y chromosome.Nature 315, 70–2.
Bishop, C. E. &Hatat, D. (1987) Molecular cloning and sequence analysis of a mouse Y chromosome transcript expressed in the testis.Nucleic Acids Research 15, 2959–69.
Bulfield, G., Siller, G. W., Wight, P. A. L. &Moore, K. J. (1984) X-chromosome-linked muscular dystrophy (mdx) in the mouse.Proc. Natl. Acad. Sci. USA 81 1189–92.
Coulton, G. R., Morgan, J. E., Partridge, T. A., &Sloper, J. C. (1988a) The mdx mouse skeletal muscle myopathy: I A histological, morphometric and biochemical investigation.Neuropathol. Appl. Neurobiol. 14, 53–70.
Coulton, G. R., Curtin, N. A., Morgan, J. E., &Partridge, T. A. (1988b) The mdx mouse skeletal muscle myopathy: contractile properties.Neuropathol. Appl. Neurobiol. 14, 299–314.
Feinberg, A. P. &Vogelstein, B. (1983) A technique for radiolabelling DNA restriction endonuclease fragments to high specific activity.Analytical Biochem. 132, 6–13.
Karpati, G., Pouilot, Y., Zubrzycka-Gaarn, E., Carpenter, S., Ray, P. N. Worton, R. G. &Holland, P. (1989) Dystrophin is expressed in mdx skeletal muscle fibers after normal myoblast implantation.Am. J. Pathol. 135, 27–32.
Lawrence, J. B. &Singer, R. H. (1985) Quantitative analysis ofin situ hybridisation for the detection of RNAs for cytoskeletal proteins.Cell 45, 407–15.
Law, P. K. Bertorini, T. E., Goodwin, T. G., Chen, M., Fang, Li, H.-J., Kirby, D. S., Florendo, H. G., Herrod, G. S. &Golden, G. S. (1990) Dystrophin production induced by myoblast transfer therapy in Duchenne Muscular DystrophyLancet July 14, 114–15.
Morgan, J. E., Coulton, G. R. &Partridge, T. A. (1987) Muscle precursor cells invade and repopulate freezekilled muscle grafts.J. Muscle Res. Cell Motil.,8, 386–96.
Morgan, J. E., Coulton, G. R &Partridge, T. A. (1989) Mdx muscle grafts retain the mdx phenotype in normal hosts.Muscle & Nerve 12, 410–19.
Morgan, J. E., Hoffman, E. P. &Partridge, T. A. (1989) Normal myogenic cells from newborn mice restore normal histology to degenerating muscles of the mdx mouse.J. Cell Biol. 111, 2437–49.
Morgan, J. E., Watt, D. J., Sloper, J. C. &Partridge T. A. (1988) Partial correction of an inherited defect of skeletal muscle by grafts of normal muscle precursor cells.J. Neurol. Sci. 86, 137–47.
Mullis, K., Faloona, F., Sharf, S., Saiki, R., Horn, G. &Erlich, H. (1986) Specific amplification of DNA in vitro: The epolymerase chain reaction.Cold Spring Harbor Symposia on Quantitative Biology 51, (1) 263–73.
Partridge, T. A., Morgan, J. E., Coulton, G. R., Hoffman, E. P. &Kunkel, L. M. (1989) Conversion of mdx myofibres from dystrophin-negative to-positive by injection of normal myoblasts.Nature 337, 176–9.
Watt, D. J., Lambert, J. E., Partridge T. A &Sloper, J. C. (1982) Incorporation of donor muscle precursor cells into an area of muscle regeneration in the host mouse.J. Neurol. Sci. 57, 319–31.
Watt, D. J., Morgan, J. E. &Partridge, T. A. (1984) Use of mononuclear precursor cells to insert allogeneic genes into growing mouse muscles.Muscle & Nerve 7, 741–50.
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Coulton, C.R., Skynner, M.J., Smith, T. et al. Localization of donor nuclei in skeletal muscle grafts byin situ hybridization to a cDNA probe. Histochem J 23, 323–327 (1991). https://doi.org/10.1007/BF01044963
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DOI: https://doi.org/10.1007/BF01044963