Vinculin is an essential component for normal myofibrillar arrangement in fetal mouse cardiac myocytes

Isao Shiraishi, David G. Simpson, Wayne Carver, Robert Price, Toshiro Hirozane, Louis Terracio, Thomas K. Borg

Research output: Contribution to journalArticle

Abstract

Vinculin is a cytoskeletal protein that is believed to be an essential component in the linkage of cytoskeletal actin filaments to the plasma membrane. To investigate the precise function of vinculin in the development of cardiac myofibrils, antisense oligodeoxynucleotides complementary to vinculin mRNA were used to perturb the expression of the protein during myofibril assembly and arrangement in mouse cardiac myocytes. Fetal (day 18-20 post-conception) mouse cardiac myocytes were isolated by collagenase digestion, separated by Percoll density gradient centrifugation, and plated on aligned collagen gels. By 72 h of culture, mouse myocytes displayed an elongated in vivo-like phenotype in parallel with the aligned fibrils of the collagen gels with polarized arrays of myofibrils. Two different antisense oligonucleotides (20-mer) altered the formation of the tissue-like phenotype of myocytes. These antisense oligonucleotides suppressed vinculin protein expression at 43.5 ± 26.8% and 48.7 ± 20.9% when compared to myocytes that were not treated. Examination of these myocytes by confocal scanning laser and transmission electron microscopy revealed a disruption of the aligned ill vivo-like phenotype, assembly of thick and thin filaments, and formulation of Z-bands. Random sequence 20-mer oligonucleotides used as controls had little detectable effect on vinculin protein expression (94.2 ± 14.8%), cell shape, normal alignment or assembly of myofibrils. These results indicate that vinculin is a critical cytoskeletal component, that functions in the determination of cell shape and the arrangement and organization of developing myofibrils.

Original languageEnglish (US)
Pages (from-to)2041-2052
Number of pages12
JournalJournal of Molecular and Cellular Cardiology
Volume29
Issue number8
DOIs
StatePublished - Aug 1997

Fingerprint

Vinculin
Cardiac Myocytes
Myofibrils
Muscle Cells
Cell Shape
Antisense Oligonucleotides
Phenotype
Collagen
Gels
Scanning Transmission Electron Microscopy
Proteins
Cytoskeletal Proteins
Density Gradient Centrifugation
Oligodeoxyribonucleotides
Collagenases
Cytoskeleton
Actin Cytoskeleton
Oligonucleotides
Digestion
Lasers

Keywords

  • Antisense oligonucleotide
  • Cardiac myocyte
  • Collagen
  • Mouse
  • Vinculin

ASJC Scopus subject areas

  • Molecular Biology
  • Cardiology and Cardiovascular Medicine

Cite this

Vinculin is an essential component for normal myofibrillar arrangement in fetal mouse cardiac myocytes. / Shiraishi, Isao; Simpson, David G.; Carver, Wayne; Price, Robert; Hirozane, Toshiro; Terracio, Louis; Borg, Thomas K.

In: Journal of Molecular and Cellular Cardiology, Vol. 29, No. 8, 08.1997, p. 2041-2052.

Research output: Contribution to journalArticle

Shiraishi, Isao ; Simpson, David G. ; Carver, Wayne ; Price, Robert ; Hirozane, Toshiro ; Terracio, Louis ; Borg, Thomas K. / Vinculin is an essential component for normal myofibrillar arrangement in fetal mouse cardiac myocytes. In: Journal of Molecular and Cellular Cardiology. 1997 ; Vol. 29, No. 8. pp. 2041-2052.
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