Focal adhesions (FAs) are specialized regions of cell attachment to the extracellular matrix. Previous works have suggested that bradykinin (BK) can modulate cell-matrix interaction. In the present study, we used a physiological cellular model to evaluate the potential role of BK in modulating FAs and stress fibers. We performed a quantitative morphometric analysis of FAs in primary cultured rat renal papillary collecting duct cells, which included size, axial ratio (shape) and average length. After 1, 5 or 10 min of incubation with BK, cultured cells were immunostained and analyzed by confocal microscopy. Although the shape of FA was not altered, BK induced a decrease in the number of vinculin-stained FAs per cell, and a decrease in both their size and their average length, but not in talin-containing FA, suggesting that BK could be inducing a restructuring of FAs. BK also induced a remodeling of the actin filament assemblies rather than their dissipation. Since we have previously demonstrated that BK stimulates activation of PLC in rat renal papillae, we tried to determine whether BK can modulate FA restructuring by this mechanism, by pre-treatment of cultured cells with the PLC inhibitor, U73122. The present study, performed under physiological conditions with cells which were not genetically manipulated, provides new experimental evidence supporting the notion that the intrarenal hormone BK modulates focal adhesions and actin cytoskeleton organization, by a mechanism that involves the activation of PLC. We propose this finding as a novel mechanism for BK modulation of the tubular collecting duct function.