title: Variation in stiffness regulates cardiac myocyte hypertrophy via signaling pathways.
abstract: Much diseased human myocardial tissue is fibrotic and stiff, which increases the work that the ventricular myocytes must perform to maintain cardiac output. The hypothesis tested is that the increased load due to greater stiffness of the substrata drives sarcomere assembly of cells, thus strengthening them. Neonatal rat ventricular myocytes (NRVM) were cultured on polyacrylamide or polydimethylsiloxane substrates with stiffness of 10 kPa, 100 kPa, or 400 kPa, or glass with stiffness of 61.9 GPa. Cell size increased with stiffness. Two signaling pathways were explored, phosphorylation of focal adhesion kinase (p-FAK) and lipids by phosphatidylinositol 4,5-bisphosphate (PIP2). Subcellular distributions of both were determined in the sarcomeric fraction by antibody localization, and total amounts were measured by Western or dot blotting, respectively. More p-FAK and PIP2 distributed to the sarcomeres of NRVM grown on stiffer substrates. Actin assembly involves the actin capping protein Z (CapZ). Both actin and CapZ dynamic exchange were significantly increased on stiffer substrates when assessed by fluorescence recovery after photobleaching (FRAP) of green fluorescent protein tags. Blunting of actin FRAP by FAK inhibition implicates linkage from mechano-signalling pathways to cell growth. Thus, increased stiffness of cardiac disease can be modeled with polymeric materials to understand how the microenvironment regulates cardiac hypertrophy.
authors.affiliation: a Department of Physiology and Biophysics, Center for Cardiovascular Research, University of Illinois at Chicago, 835 S. Wolcott Ave, Chicago, IL 60612, USA.
authors.forename: Jieli
authors.initials: J
authors.lastname: Li
authors.affiliation: b Department of Bioengineering, University of Illinois at Chicago, 851 South Morgan Street, Chicago, IL 60607, USA.
authors.forename: Michael A
authors.initials: MA
authors.lastname: Mkrtschjan
authors.affiliation: a Department of Physiology and Biophysics, Center for Cardiovascular Research, University of Illinois at Chicago, 835 S. Wolcott Ave, Chicago, IL 60612, USA.
authors.forename: Ying-Hsi
authors.initials: YH
authors.lastname: Lin
authors.affiliation: a Department of Physiology and Biophysics, Center for Cardiovascular Research, University of Illinois at Chicago, 835 S. Wolcott Ave, Chicago, IL 60612, USA.|b Department of Bioengineering, University of Illinois at Chicago, 851 South Morgan Street, Chicago, IL 60607, USA.
authors.forename: Brenda
authors.initials: B
authors.lastname: Russell
keywords: actin assembly; assemblage de l’actine; focal adhesion kinase; lipid signaling; mechano-transduction; mécano-transduction; rigidité du substrat; signalisation lipidique; substrate stiffness; « focal adhesion kinase »