title: Externally applied cyclic strain regulates localization of focal contact components in cultured smooth muscle cells.
abstract: Mechanical signals are critical regulators of cellular gene expression, yet little is understood of the mechanism whereby cells sense mechanical forces. In this study we have tested the hypothesis that mechanical strain applied to populations of cells via their adhesion substrate rapidly alters the cellular distribution of focal contact proteins. Focal contact-associated components (vinculin, a-actinin, paxillin) were assayed by immunofluorescence microscopy and quantitative western blotting. Application of a single step increase in strain in multiple experiments caused overall a small change in focal contact-associated vinculin. In contrast, cyclic strain induced a large and very reproducible increase in detergent-insoluble vinculin (52% relative to static) after just 1 min of strain. Insoluble paxillin was transiently enriched with a similar time course, whereas insoluble a-actinin did not change significantly in response to cyclic strain. Rhodamine-labeled chicken vinculin added to permeabilized cells preferentially localized to focal contacts in response to cyclic strain, but not a single step increase in strain. These findings establish that insoluble levels of focal contact components are altered rapidly following application of an appropriate number of mechanical perturbations, and suggest that at least one component of the mechanism does not involve soluble intermediates.
authors.affiliation: Department of Chemical Engineering, The University of Michigan, Ann Arbor 48109, USA.
authors.forename: James J
authors.initials: JJ
authors.lastname: Cunningham
authors.forename: Jennifer J
authors.initials: JJ
authors.lastname: Linderman
authors.forename: David J
authors.initials: DJ
authors.lastname: Mooney
mesh_terms: D000185:Actinin / Q000502:physiology; D000818:Animals; D001013:Aorta, Thoracic / Q000502:physiology; D002448:Cell Adhesion / Q000502:physiology*; D015815:Cell Adhesion Molecules / Q000502:physiology; D002478:Cells, Cultured; D003598:Cytoskeletal Proteins / Q000502:physiology*; D022001:Focal Adhesions / Q000502:physiology; D008297:Male; D040542:Mechanotransduction, Cellular / Q000502:physiology*; D009131:Muscle, Smooth, Vascular / Q000166:cytology* / Q000502:physiology*; D051419:Paxillin; D010507:Periodicity; D010750:Phosphoproteins / Q000502:physiology; D051381:Rats; D011917:Rats, Inbred Lew; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013314:Stress, Mechanical; D016596:Vinculin / Q000502:physiology
chemical_list: D015815:Cell Adhesion Molecules; D003598:Cytoskeletal Proteins; D051419:Paxillin; D010750:Phosphoproteins; C496063:Pxn protein, rat; D000185:Actinin; D016596:Vinculin