Localization of donor nuclei in skeletal muscle grafts by in situ hybridization to a cDNA probe.
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-radioactive in situ hybridization of a Y 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.
Coulton
G R
GR
Department of Biochemistry, Charing Cross and Westminster Medical School, London, UK.
Skynner
M J
MJ
Smith
T
T
Pagel
C N
CN
Partridge
T A
TA
D000818:Animals; D001483:Base Sequence; D002467:Cell Nucleus / Q000737:chemistry; D015342:DNA Probes / Q000235:genetics; D004076:Digoxigenin; D016189:Dystrophin / Q000032:analysis*; D005260:Female; D005956:Glucose-6-Phosphate Isomerase / Q000032:analysis; D007150:Immunohistochemistry; D007525:Isoelectric Focusing; D008297:Male; D051379:Mice; D008819:Mice, Nude; D008969:Molecular Sequence Data; D009132:Muscles / Q000166:cytology* / Q000637:transplantation; D009137:Muscular Dystrophy, Animal / Q000473:pathology*; D009693:Nucleic Acid Hybridization; D051381:Rats
D015342:DNA Probes; D016189:Dystrophin; D005956:Glucose-6-Phosphate Isomerase; D004076:Digoxigenin