title: Inclusion formation in Huntington's disease R6/2 mouse muscle cultures.
abstract: Huntington's disease (HD) is an autosomal dominant disorder caused by an expansion in the number of glutamine repeats in the N-terminal region of the huntingtin protein. Nuclear and cytoplasmic aggregates of the N-terminal portion of huntingtin have been found in the brains of HD patients and the brains and non-neuronal tissues of the R6/2 HD transgenic mouse. We have cultured myoblasts and myotubes from transgenic R6/2 mice and littermate controls to investigate the formation of these inclusions in post mitotic cells. Huntingtin immunoreactivity was intense in differentiating, desmin positive myoblasts and myotubes from both control and R6/2 mice suggesting that it may play a role in myotube differentiation. Following differentiation huntingtin and ubiquitin positive aggregates were observed in R6/2 but not control cultures. After 3 weeks in differentiation medium cytoplasmic huntingtin and ubiquitin immunoreactive aggregates were observed in non-myotube cells, while nuclear huntingtin aggregates were seen in a proportion of myotubes after 6 weeks. Growth in the absence of serum resulted in a marked increase in the number of R6/2 myotubes containing nuclear inclusions after 6 weeks demonstrating that environmental factors influenced huntingtin aggregate formation in these cells. Consequently, cultured myotubes from R6/2 mice may be a useful post mitotic cell culture model to study both the biochemical consequences of huntingtin aggregates and the factors that may influence aggregate formation.
authors.affiliation: University Department of Clinical Neurosciences, Royal Free and University College Medical College, University College London, UK.
authors.forename: M
authors.initials: M
authors.lastname: Orth
authors.forename: J M
authors.initials: JM
authors.lastname: Cooper
authors.forename: G P
authors.initials: GP
authors.lastname: Bates
authors.forename: A H V
authors.initials: AH
authors.lastname: Schapira
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D002478:Cells, Cultured; D003893:Desmin / Q000096:biosynthesis; D004195:Disease Models, Animal; D000071058:Huntingtin Protein; D006816:Huntington Disease / Q000473:pathology*; D002479:Inclusion Bodies / Q000473:pathology*; D051379:Mice; D008822:Mice, Transgenic; D018485:Muscle Fibers, Skeletal / Q000166:cytology / Q000378:metabolism / Q000473:pathology*; D032446:Myoblasts / Q000166:cytology / Q000378:metabolism / Q000473:pathology*; D009419:Nerve Tissue Proteins / Q000096:biosynthesis; D009687:Nuclear Proteins / Q000096:biosynthesis
chemical_list: D003893:Desmin; C506812:Htt protein, mouse; D000071058:Huntingtin Protein; D009419:Nerve Tissue Proteins; D009687:Nuclear Proteins