Tension development during contractile stimulation of smooth muscle requires recruitment of paxillin and vinculin to the membrane

Anabelle Opazo Saez, Wenwu Zhang, Yidi Wu, Christopher E. Turner, Dale D. Tang, Susan J. Gunst

Research output: Contribution to journalArticle

97 Scopus citations

Abstract

Cytoskeletal reorganization of the smooth muscle cell in response to contractile stimulation may be an important fundamental process in regulation of tension development. We used confocal microscopy to analyze the effects of cholinergic stimulation on localization of the cytoskeletal proteins vinculin, paxillin, talin and focal adhesion kinase (FAK) in freshly dissociated tracheal smooth muscle cells. All four proteins were localized at the membrane and throughout the cytoplasm of unstimulated cells, but their concentration at the membrane was greater in acetylcholine (ACh)-stimulated cells. Antisense oligonucleotides were introduced into tracheal smooth muscle tissues to deplete paxillin protein, which also inhibited contraction in response to ACh. In cells dissociated from paxillin-depleted muscle tissues, redistribution of vinculin to the membrane in response to ACh was prevented, but redistribution of FAK and talin was not inhibited. Muscle tissues were transfected with plasmids encoding a paxillin mutant containing a deletion of the LIM3 domain (paxillin LIM3 dl 444-494), the primary determinant for targeting paxillin to focal adhesions. Expression of paxillin LIM3 dl in muscle tissues also inhibited contractile force and prevented cellular redistribution of paxillin and vinculin to the membrane in response to ACh, but paxillin LIM3 dl did not inhibit increases in intracellular Ca2+ or myosin light chain phosphorylation. Our results demonstrate that recruitment of paxillin and vinculin to smooth muscle membrane is necessary for tension development and that recruitment of vinculin to the membrane is regulated by paxillin. Vinculin and paxillin may participate in regulating the formation of linkages between the cytoskeleton and integrin proteins that mediate tension transmission between the contractile apparatus and the extracellular matrix during smooth muscle contraction.

Original languageEnglish (US)
Pages (from-to)C433-C447
JournalAmerican Journal of Physiology - Cell Physiology
Volume286
Issue number2 55-2
StatePublished - Feb 1 2004

Keywords

  • Cytoskeleton
  • Immunofluorescence
  • Plasmids
  • Talin
  • Tissue transfection

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Cell Biology
  • Physiology

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