title: Muscle dysfunction and structural defects of dystrophin-null sapje mutant zebrafish larvae are rescued by ataluren treatment.
abstract: Sapje zebrafish carry a mutation in the dystrophin gene, which results in a premature stop codon, and a severe muscle phenotype. They display several of the structural characteristics of Duchenne muscular dystrophy (DMD). Ataluren (PTC124) is proposed to cause readthrough of premature stop codons and has been introduced as a potential treatment of genetic disorders. Clinical trials in DMD have shown promise, although with complex dose dependency. We have established physiology techniques, enabling high resolution of contractile function in skeletal muscle of zebrafish larvae. We aimed to provide a mechanical analysis of sapje larval muscle and examine effects of ataluren. Homozygous 5 d postfertilization (dpf) sapje larvae exhibited structural defects with 50% decrease in active tension. Ataluren (0.1-1 μM, 3-5 dpf) improved contractile function (~60% improvement of force at 0.5 μM) and dystrophin expression. Controls were not affected. Higher doses (5 μM, 35 μM) impaired contractile function, an effect also observed in controls, suggesting unspecific negative effects at high concentrations. In summary, Sapje larvae exhibit impaired contractile performance and provide a relevant DMD model for functional studies. Ataluren significantly improves skeletal muscle function in the sapje larvae, most likely reflecting an observed increase in dystrophin expression. The bell-shaped dose dependence in sapje resembles that previously reported in clinical DMD studies.
authors.affiliation: 1 Department of Physiology and Pharmacology, Karolinska Institutet, Von Eulers v 8, SE 171 77 Stockholm, Sweden. anders.arner@ki.se.
authors.forename: Mei
authors.initials: M
authors.lastname: Li
authors.forename: Monika
authors.initials: M
authors.lastname: Andersson-Lendahl
authors.forename: Thomas
authors.initials: T
authors.lastname: Sejersen
authors.forename: Anders
authors.initials: A
authors.lastname: Arner
mesh_terms: D000818:Animals; D015153:Blotting, Western; D004305:Dose-Response Relationship, Drug; D016189:Dystrophin / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D007814:Larva / Q000187:drug effects / Q000235:genetics / Q000502:physiology; D018482:Muscle, Skeletal / Q000187:drug effects / Q000378:metabolism / Q000503:physiopathology; D009137:Muscular Dystrophy, Animal / Q000235:genetics* / Q000378:metabolism / Q000503:physiopathology; D020388:Muscular Dystrophy, Duchenne / Q000235:genetics / Q000378:metabolism / Q000503:physiopathology; D009154:Mutation; D010069:Oxadiazoles / Q000494:pharmacology*; D013314:Stress, Mechanical; D015027:Zebrafish / Q000235:genetics* / Q000378:metabolism; D029961:Zebrafish Proteins / Q000235:genetics* / Q000378:metabolism
chemical_list: D016189:Dystrophin; D010069:Oxadiazoles; D029961:Zebrafish Proteins; C515878:ataluren
keywords: Danio rerio; Duchenne muscular dystrophy; skeletal muscle; therapy