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Am J Physiol Renal Physiol 288: F133-F141, 2005. First published September 14, 2004; doi:10.1152/ajprenal.00238.2004
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Transepithelial pressure pulses induce nucleotide release in polarized MDCK cells

H. A. Praetorius,1,2 J. Frøkiær,1,2 and J. Leipziger2,3

1Clinical Institute, 3Institute of Physiology, and 2The Water and Salt Research Center, University of Aarhus, Aarhus, Denmark

Submitted 28 June 2004 ; accepted in final form 9 September 2004

The release of nucleotides is involved in mechanosensation in various epithelial cells. Intriguingly, kidney epithelial cells are absolutely dependent on the primary cilium to sense changes in apical laminar flow. During fluid passage, the renal epithelial cells are subjected to various mechanical stimuli in addition to changes in the laminar flow rate. In the distal part of the collecting duct, the epithelial cells are exposed to pressure changes and possibly distension during papillary contractions. The aim of the present study was to determine whether nucleotide release contributes to mechanosensation in kidney epithelial cells, thereby establishing whether pressure changes are sufficient to produce nucleotide-mediated responses. Madin-Darby canine kidney (MDCK) cells grown on permeable supports were mounted in a closed double perfusion chamber on an inverted microscope. The intracellular Ca2+ concentration ([Ca2+]i) was monitored with the Ca2+-sensitive fluorescence probe fluo 4. Transepithelial pressure pulses of 30–80 mmHg produced a transient increase in [Ca2+]i of MDCK cells. This response is independent of the primary cilium, since it is readily observed in immature cells that do not yet express primary cilia. The amplitudes of the pressure-induced Ca2+ transients varied with the applied chamber pressure in a quantity-dependent manner. The ATPase apyrase and the P2Y antagonist suramin significantly reduced the pressure-induced Ca2+ transients. Applying apyrase or suramin to both sides of the preparation simultaneously nearly abolished the pressure-induced Ca2+ response. In conclusion, these observations suggest that rapid pressure changes induce both apical and basolateral nucleotide release that contribute to mechanosensation in kidney epithelial cells.

ATP; P2Y; calcium; mechanosensation


Address for reprint requests and other correspondence: H. A. Praetorius, The Water and Salt Research Ctr., Clinical Institute, Brendstrupgaardsvej 1, 8200 Aarhus N, Denmark (helle.praetorius{at}ki.au.dk)




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