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Title: Characterization of zebrafish dysferlin by morpholino knockdown

Abstract

Highlights: {yields} cDNAs of zebrafish dysferlin were cloned (6.3 kb). {yields} The dysferlin expression was detected in skeletal muscle, heart and eye. {yields} Injection of antisense morpholinos to dysferlin caused marked muscle disorganization. {yields} Zebrafish dysferlin expression may be involved in stabilizing muscle structures. -- 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 wasmore » shown to be reduced at the myosepta of zebrafish muscle using immunohistochemistry, although the expression of other muscle membrane components, dystrophin, laminin, {beta}-dystroglycan were detected normally. Our data suggest that zebrafish dysferlin expression is involved in stabilizing muscle structures and its downregulation causes muscle disorganization.« less

Authors:
; ;  [1];  [2];  [2];  [1];  [1];  [2];  [2]
  1. Division of Genetics, Program in Genomics, Children's Hospital Boston, MA (United States)
  2. (United States)
Publication Date:
OSTI Identifier:
22207508
Resource Type:
Journal Article
Journal Name:
Biochemical and Biophysical Research Communications
Additional Journal Information:
Journal Volume: 413; Journal Issue: 2; Other Information: Copyright (c) 2011 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved.; Country of input: International Atomic Energy Agency (IAEA); Journal ID: ISSN 0006-291X
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; AMINO ACID SEQUENCE; ANTIBODIES; BIREFRINGENCE; DISEASES; FERTILIZATION; GENES; HEART; INJECTION; LIMBS; MESSENGER-RNA; MUSCLES; MUTATIONS; OLIGONUCLEOTIDES; PHENOTYPE

Citation Formats

Kawahara, Genri, Serafini, Peter R., Myers, Jennifer A., Department of Genetics, Harvard Medical School, MA, The Manton Center for Orphan Disease Research, Children's Hospital Boston, MA, Alexander, Matthew S., Kunkel, Louis M., E-mail: kunkel@enders.tch.harvard.edu, Department of Genetics, Harvard Medical School, MA, and The Manton Center for Orphan Disease Research, Children's Hospital Boston, MA. Characterization of zebrafish dysferlin by morpholino knockdown. United States: N. p., 2011. Web. doi:10.1016/J.BBRC.2011.08.105.
Kawahara, Genri, Serafini, Peter R., Myers, Jennifer A., Department of Genetics, Harvard Medical School, MA, The Manton Center for Orphan Disease Research, Children's Hospital Boston, MA, Alexander, Matthew S., Kunkel, Louis M., E-mail: kunkel@enders.tch.harvard.edu, Department of Genetics, Harvard Medical School, MA, & The Manton Center for Orphan Disease Research, Children's Hospital Boston, MA. Characterization of zebrafish dysferlin by morpholino knockdown. United States. doi:10.1016/J.BBRC.2011.08.105.
Kawahara, Genri, Serafini, Peter R., Myers, Jennifer A., Department of Genetics, Harvard Medical School, MA, The Manton Center for Orphan Disease Research, Children's Hospital Boston, MA, Alexander, Matthew S., Kunkel, Louis M., E-mail: kunkel@enders.tch.harvard.edu, Department of Genetics, Harvard Medical School, MA, and The Manton Center for Orphan Disease Research, Children's Hospital Boston, MA. Fri . "Characterization of zebrafish dysferlin by morpholino knockdown". United States. doi:10.1016/J.BBRC.2011.08.105.
@article{osti_22207508,
title = {Characterization of zebrafish dysferlin by morpholino knockdown},
author = {Kawahara, Genri and Serafini, Peter R. and Myers, Jennifer A. and Department of Genetics, Harvard Medical School, MA and The Manton Center for Orphan Disease Research, Children's Hospital Boston, MA and Alexander, Matthew S. and Kunkel, Louis M., E-mail: kunkel@enders.tch.harvard.edu and Department of Genetics, Harvard Medical School, MA and The Manton Center for Orphan Disease Research, Children's Hospital Boston, MA},
abstractNote = {Highlights: {yields} cDNAs of zebrafish dysferlin were cloned (6.3 kb). {yields} The dysferlin expression was detected in skeletal muscle, heart and eye. {yields} Injection of antisense morpholinos to dysferlin caused marked muscle disorganization. {yields} Zebrafish dysferlin expression may be involved in stabilizing muscle structures. -- 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, {beta}-dystroglycan were detected normally. Our data suggest that zebrafish dysferlin expression is involved in stabilizing muscle structures and its downregulation causes muscle disorganization.},
doi = {10.1016/J.BBRC.2011.08.105},
journal = {Biochemical and Biophysical Research Communications},
issn = {0006-291X},
number = 2,
volume = 413,
place = {United States},
year = {2011},
month = {9}
}