Bradykinin modulates focal adhesions and induces stress fiber remodeling

MARÍA GABRIELA MARQUEZ; MARIA C. FERNÁNDEZ TOME; NICOLAS O. FAVALE; LUCILA G PESCIO; NORMA STERIN-SPEZIALE

AMERICAN JOURNAL OF PHYSIOLOGY-RENAL PHYSIOLOGY - (Print)

APS

Año: 2008 vol. 294 p. 603 - 613

0363-6127

Ma´rquez MG, del Carmen Ferna´ndez-Tome M, Favale NO, Pescio LG, Sterin-Speziale NB. Bradykinin modulates focal adhesions and induces stress fiber remodeling in renal papillary collecting duct cells. Am J Physiol Renal Physiol 294: F603–F613, 2008. First published December 26, 2007; doi:10.1152/ajprenal.00234.2007.— 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 FAs 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 FAs, thus 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 attempted to determine whether BK can modulate FA restructuring by this mechanism, by pretreating cultured cells with the PLC inhibitor U73122. The present study, performed under physiological conditions with cells that were not genetically manipulated, provides new experimental evidence supporting the notion that the intrarenal hormone BK modulates FAs and actin cytoskeleton organization through a mechanism that involves the activation of PLC. We propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. under physiological conditions with cells that were not genetically manipulated, provides new experimental evidence supporting the notion that the intrarenal hormone BK modulates FAs and actin cytoskeleton organization through a mechanism that involves the activation of PLC. We propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. modulate FA restructuring by this mechanism, by pretreating cultured cells with the PLC inhibitor U73122. The present study, performed under physiological conditions with cells that were not genetically manipulated, provides new experimental evidence supporting the notion that the intrarenal hormone BK modulates FAs and actin cytoskeleton organization through a mechanism that involves the activation of PLC. We propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. under physiological conditions with cells that were not genetically manipulated, provides new experimental evidence supporting the notion that the intrarenal hormone BK modulates FAs and actin cytoskeleton organization through a mechanism that involves the activation of PLC. We propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. published December 26, 2007; doi:10.1152/ajprenal.00234.2007.— 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 FAs 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 FAs, thus 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 attempted to determine whether BK can modulate FA restructuring by this mechanism, by pretreating cultured cells with the PLC inhibitor U73122. The present study, performed under physiological conditions with cells that were not genetically manipulated, provides new experimental evidence supporting the notion that the intrarenal hormone BK modulates FAs and actin cytoskeleton organization through a mechanism that involves the activation of PLC. We propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. under physiological conditions with cells that were not genetically manipulated, provides new experimental evidence supporting the notion that the intrarenal hormone BK modulates FAs and actin cytoskeleton organization through a mechanism that involves the activation of PLC. We propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. modulate FA restructuring by this mechanism, by pretreating cultured cells with the PLC inhibitor U73122. The present study, performed under physiological conditions with cells that were not genetically manipulated, provides new experimental evidence supporting the notion that the intrarenal hormone BK modulates FAs and actin cytoskeleton organization through a mechanism that involves the activation of PLC. We propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. under physiological conditions with cells that were not genetically manipulated, provides new experimental evidence supporting the notion that the intrarenal hormone BK modulates FAs and actin cytoskeleton organization through a mechanism that involves the activation of PLC. We propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. and induces stress fiber remodeling in renal papillary collecting duct cells. Am J Physiol Renal Physiol 294: F603–F613, 2008. First published December 26, 2007; doi:10.1152/ajprenal.00234.2007.— 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 FAs 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 FAs, thus 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 attempted to determine whether BK can modulate FA restructuring by this mechanism, by pretreating cultured cells with the PLC inhibitor U73122. The present study, performed under physiological conditions with cells that were not genetically manipulated, provides new experimental evidence supporting the notion that the intrarenal hormone BK modulates FAs and actin cytoskeleton organization through a mechanism that involves the activation of PLC. We propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. under physiological conditions with cells that were not genetically manipulated, provides new experimental evidence supporting the notion that the intrarenal hormone BK modulates FAs and actin cytoskeleton organization through a mechanism that involves the activation of PLC. We propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. modulate FA restructuring by this mechanism, by pretreating cultured cells with the PLC inhibitor U73122. The present study, performed under physiological conditions with cells that were not genetically manipulated, provides new experimental evidence supporting the notion that the intrarenal hormone BK modulates FAs and actin cytoskeleton organization through a mechanism that involves the activation of PLC. We propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. under physiological conditions with cells that were not genetically manipulated, provides new experimental evidence supporting the notion that the intrarenal hormone BK modulates FAs and actin cytoskeleton organization through a mechanism that involves the activation of PLC. We propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. published December 26, 2007; doi:10.1152/ajprenal.00234.2007.— 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 FAs 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 FAs, thus 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 attempted to determine whether BK can modulate FA restructuring by this mechanism, by pretreating cultured cells with the PLC inhibitor U73122. The present study, performed under physiological conditions with cells that were not genetically manipulated, provides new experimental evidence supporting the notion that the intrarenal hormone BK modulates FAs and actin cytoskeleton organization through a mechanism that involves the activation of PLC. We propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. under physiological conditions with cells that were not genetically manipulated, provides new experimental evidence supporting the notion that the intrarenal hormone BK modulates FAs and actin cytoskeleton organization through a mechanism that involves the activation of PLC. We propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. modulate FA restructuring by this mechanism, by pretreating cultured cells with the PLC inhibitor U73122. The present study, performed under physiological conditions with cells that were not genetically manipulated, provides new experimental evidence supporting the notion that the intrarenal hormone BK modulates FAs and actin cytoskeleton organization through a mechanism that involves the activation of PLC. We propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. under physiological conditions with cells that were not genetically manipulated, provides new experimental evidence supporting the notion that the intrarenal hormone BK modulates FAs and actin cytoskeleton organization through a mechanism that involves the activation of PLC. We propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. Bradykinin modulates focal adhesions and induces stress fiber remodeling in renal papillary collecting duct cells. Am J Physiol Renal Physiol 294: F603–F613, 2008. First published December 26, 2007; doi:10.1152/ajprenal.00234.2007.— 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 FAs 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 FAs, thus 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 attempted to determine whether BK can modulate FA restructuring by this mechanism, by pretreating cultured cells with the PLC inhibitor U73122. The present study, performed under physiological conditions with cells that were not genetically manipulated, provides new experimental evidence supporting the notion that the intrarenal hormone BK modulates FAs and actin cytoskeleton organization through a mechanism that involves the activation of PLC. We propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. under physiological conditions with cells that were not genetically manipulated, provides new experimental evidence supporting the notion that the intrarenal hormone BK modulates FAs and actin cytoskeleton organization through a mechanism that involves the activation of PLC. We propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. modulate FA restructuring by this mechanism, by pretreating cultured cells with the PLC inhibitor U73122. The present study, performed under physiological conditions with cells that were not genetically manipulated, provides new experimental evidence supporting the notion that the intrarenal hormone BK modulates FAs and actin cytoskeleton organization through a mechanism that involves the activation of PLC. We propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. under physiological conditions with cells that were not genetically manipulated, provides new experimental evidence supporting the notion that the intrarenal hormone BK modulates FAs and actin cytoskeleton organization through a mechanism that involves the activation of PLC. We propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. published December 26, 2007; doi:10.1152/ajprenal.00234.2007.— 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 FAs 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 FAs, thus 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 attempted to determine whether BK can modulate FA restructuring by this mechanism, by pretreating cultured cells with the PLC inhibitor U73122. The present study, performed under physiological conditions with cells that were not genetically manipulated, provides new experimental evidence supporting the notion that the intrarenal hormone BK modulates FAs and actin cytoskeleton organization through a mechanism that involves the activation of PLC. We propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. under physiological conditions with cells that were not genetically manipulated, provides new experimental evidence supporting the notion that the intrarenal hormone BK modulates FAs and actin cytoskeleton organization through a mechanism that involves the activation of PLC. We propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. modulate FA restructuring by this mechanism, by pretreating cultured cells with the PLC inhibitor U73122. The present study, performed under physiological conditions with cells that were not genetically manipulated, provides new experimental evidence supporting the notion that the intrarenal hormone BK modulates FAs and actin cytoskeleton organization through a mechanism that involves the activation of PLC. We propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. under physiological conditions with cells that were not genetically manipulated, provides new experimental evidence supporting the notion that the intrarenal hormone BK modulates FAs and actin cytoskeleton organization through a mechanism that involves the activation of PLC. We propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. Am J Physiol Renal Physiol 294: F603–F613, 2008. First published December 26, 2007; doi:10.1152/ajprenal.00234.2007.— 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 FAs 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 FAs, thus 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 attempted to determine whether BK can modulate FA restructuring by this mechanism, by pretreating cultured cells with the PLC inhibitor U73122. The present study, performed under physiological conditions with cells that were not genetically manipulated, provides new experimental evidence supporting the notion that the intrarenal hormone BK modulates FAs and actin cytoskeleton organization through a mechanism that involves the activation of PLC. We propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. under physiological conditions with cells that were not genetically manipulated, provides new experimental evidence supporting the notion that the intrarenal hormone BK modulates FAs and actin cytoskeleton organization through a mechanism that involves the activation of PLC. We propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. modulate FA restructuring by this mechanism, by pretreating cultured cells with the PLC inhibitor U73122. The present study, performed under physiological conditions with cells that were not genetically manipulated, provides new experimental evidence supporting the notion that the intrarenal hormone BK modulates FAs and actin cytoskeleton organization through a mechanism that involves the activation of PLC. We propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. under physiological conditions with cells that were not genetically manipulated, provides new experimental evidence supporting the notion that the intrarenal hormone BK modulates FAs and actin cytoskeleton organization through a mechanism that involves the activation of PLC. We propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. in rat renal papillae, we attempted to determine whether BK can modulate FA restructuring by this mechanism, by pretreating cultured cells with the PLC inhibitor U73122. The present study, performed under physiological conditions with cells that were not genetically manipulated, provides new experimental evidence supporting the notion that the intrarenal hormone BK modulates FAs and actin cytoskeleton organization through a mechanism that involves the activation of PLC. We propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. under physiological conditions with cells that were not genetically manipulated, provides new experimental evidence supporting the notion that the intrarenal hormone BK modulates FAs and actin cytoskeleton organization through a mechanism that involves the activation of PLC. We propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. inhibitor U73122. The present study, performed under physiological conditions with cells that were not genetically manipulated, provides new experimental evidence supporting the notion that the intrarenal hormone BK modulates FAs and actin cytoskeleton organization through a mechanism that involves the activation of PLC. We propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. . We propose this finding as a novel mechanism for BK modulation of tubular collecting duct function.