RT Journal Article SR Electronic T1 REGULATION OF ACTIN CYTOSKELETON AND FOCAL ADHESION DYNAMICS BY INTERNAL AND EXTERNAL FORCES IN VASCULAR SMOOTH MUSCLE CELLS JF Heart JO Heart FD BMJ Publishing Group Ltd and British Cardiovascular Society SP A4 OP A5 DO 10.1136/heartjnl-2014-306916.12 VO 100 IS Suppl 4 A1 M Pieri A1 A Mekhalfia A1 M Platt A1 J Ohanian A1 V Ohanian YR 2014 UL http://heart.bmj.com/content/100/Suppl_4/A4.3.abstract AB Introduction Aberrant sensing and transmission of force by vascular smooth muscle cells (VSMC) results in vascular remodelling, increased arterial stiffness and hypertension. Actin cytoskeleton (CK) remodelling is essential for VSMC responses to vasoconstrictors and mechanical stimuli. Focal adhesions (FAs) link the extracellular matrix and the actin CK allowing sensing and transmission of force. Hic-5 is highly expressed in VSMC and regulates FA assembly and actin CK remodelling in arteries. Aims To investigate whether alterations in intracellular force generation (actin CK remodelling) or extracellular forces (substrate stiffness) regulate FA dynamics in VSMC. Methods To induce actin CK remodelling rVSMC were stimulated with Noradrenaline (NA; 1.5 µM or 15 µM) or treated with blebbistatin (myosin II inhibitor, 50 µM). To alter extracellular stiffness rVSMC were cultured on substrates of varying stiffness; soft, 2.61±0.82 kPa; stiff, 40.40±2.39 kPa; glass- 50–90 GPa. Hic-5 and actin filament distribution were determined using immunofluorescence. Results NA induced a decrease in Hic-5 in FAs (p≤0.01) and redistribution of actin filaments to the cell periphery. Blebbistatin also induced a decrease in Hic-5 in FAs and actin filament redistribution to the cell periphery (p≤0.0001). Hic-5 in FAs increased (p≤0.0001) and actin CK remodelling occurred with increasing substrate stiffness. Conclusion Actin CK and FA structure is dynamically regulated in response to alterations in intra- and extracellular forces in VSMC. Hic-5 redistribution in response to vasoconstrictor and mechanical stimuli implicates Hic-5 in FA remodelling and mechanosensing in VSMC. Altered Hic-5 function may contribute to arterial remodelling in cardiovascular disease.