PUBLICATION

Characterization of zebrafish dysferlin by morpholino knockdown

Authors
Kawahara, G., Serafini, P.R., Myers, J.A., Alexander, M.S., and Kunkel, L.M.
ID
ZDB-PUB-110907-39
Date
2011
Source
Biochemical and Biophysical Research Communications   413(2): 358-63 (Journal)
Registered Authors
Kawahara, Genri, Kunkel, Louis M.
Keywords
dysferlin, zebrafish, morpholino
MeSH Terms
  • Amino Acid Sequence
  • Animals
  • Cloning, Molecular
  • DNA, Complementary/genetics
  • Dystroglycans/metabolism
  • Dystrophin/metabolism
  • Gene Knockdown Techniques
  • Laminin/metabolism
  • Membrane Proteins/genetics
  • Membrane Proteins/metabolism*
  • Molecular Sequence Data
  • Morpholinos/genetics
  • Muscle, Skeletal/abnormalities*
  • Muscle, Skeletal/metabolism*
  • Mutation
  • Zebrafish/embryology*
  • Zebrafish/genetics
  • Zebrafish/metabolism
  • Zebrafish Proteins/genetics
  • Zebrafish Proteins/metabolism*
PubMed
21893049 Full text @ Biochem. Biophys. Res. Commun.
Abstract
Mutations in the gene encoding dysferlin cause two distinct muscular dystrophy phenotypes: limb-girdle muscular dystrophy type 2B (LGMD-2B) and Miyoshi myopathy (MM). Dysferlin is a large transmembrane protein involved in myoblast fusion and membrane resealing. Zebrafish represent an ideal animal model to use for studying muscle disease including abnormalities of dysferlin. cDNAs of zebrafish dysferlin were cloned (6.3 kb) and the predicted amino acid sequences, showed 68% similarity to predicted amino acid sequences of mammalian dysferlin. The expression of dysferlin was mainly in skeletal muscle, heart and eye, and the expression could be detected as early as 11 h post fertilization (hpf). Three different antisense oligonucleotide morpholinos were targeted to inhibit translation of this dysferlin mRNA and the morpholino-injected fish showed marked muscle disorganization which could be detected by birefringence assay. Western blot analysis using dysferlin antibodies showed that the expression of dysferlin was reduced in each of the three morphants. Dysferlin expression was shown to be reduced at the myosepta of zebrafish muscle using immunohistochemistry, although the expression of other muscle membrane components, dystrophin, laminin, β-dystroglycan were detected normally. Our data suggest that zebrafish dysferlin expression is involved in stabilizing muscle structures and its downregulation causes muscle disorganization.
Genes / Markers
Figures
Show all Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping