The ability of vascular smooth muscle cells (VSMCs) to regulate vascular caliber is anchorage-dependent in that cells require physical attachment to the extracellular matrix (ECM) and to neighboring cells in order to perform the necessary mechanical work. Importantly, modulation of this vasoregulatory property of VSMCs would significantly impact autoregulation of tissue blood flow as well as control of peripheral vascular resistance. Integrin-mediated attachment of VSMCs to ECM proteins provides a site of attachment that physically creates a functional mechanical axis with the cytoskeleton for bi-directionally transmitting mechanical forces and generation of biochemical cell signals through inside-out and outside-in mechanisms. In previous work, we have demonstrated that integrins play a role in modulating vascular tone and are important for regulation of arteriolar diameter and the vascular myogenic response supporting specific involvement in signaling related to contractile behavior. In other laboratories, biochemical evidence indicates that smooth muscle cell contraction is accompanied by rapid parallel remodeling of the actin cytoskeleton that occurs coincident with the development of tone. This remodeling is hypothesized to function to strengthen the contractile axis for transmission of force. Questions remain whether or not this type of actin remodeling characterizes the responses of VSMCs from resistance arterioles and if this type of remodeling process occurs concurrently with contractile changes in cell shape and whether it is synchronized with integrin adhesion. Enhanced integrin adhesion would be expected to also assist in bearing the increased mechanical loads associated with contractile activation. In these studies we have used atomic force microscopy (AFM) as a biomechanical method for real-time monitoring of cell contraction, cortical elasticity and adhesion to test the hypothesis that the integrins and their interactions with the ECM are enhanced and coordinated with contractile activation in VSMCs. The AFM allows this hypothesis to be biomechanically tested such that events related to contractile activation (cell movement), cytoskeletal remodeling (cell elasticity) and adhesion could be mechanically assessed and simultaneously measured. VSMCs were isolated from skeletal muscle arterioles and studied using AFM probes with tips that were bio-functionalized with the ECM protein fibronectin (FN). A nano-indentation protocol was used in which AFM probes were repeatedly interacted with the cell surface. The AFM approach curve provided information on cell height (contact point) and cell elasticity (Young’s modulus) whereas cell adhesion was measured by evaluating adhesive interactions in the retraction curve. AFM measurements were continuously recorded from VSMC before and after exposure of the cells to the vasoconstrictor angiotensin II (10-7 M). The results demonstrated that in response to AII there was an immediate rise in cell height indicating cell contraction. Cell cortical elasticity was also observed to increase but changed more slowly and lagged behind the cell height change. Increased adhesion to FN was rapidly detected following addition of AII and appeared to follow the change in cell height and precede the change in elasticity. Increased adhesion was apparent as an increased unbinding force and increased probability of binding between the AFM probe and the VSMC. These results support the hypothesis that control of VSMC adhesion to the ECM is dynamically coordinated with VSMC contractile activation and a change in cell elasticity. The parallel linking of integrin adhesion with contractile activation and cytoskeletal remodeling is potentially important to expanding our current framework of understanding of resistance artery function. Support to GAM NIH1P01HL095486.