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Am J Physiol Renal Physiol (September 14, 2004). doi:10.1152/ajprenal.00238.2004
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Submitted on June 28, 2004
Accepted on September 9, 2004

Transepithelial pressure pulses induce nucleotide release in polarized MDCK cells

H. A. Praetorius1*, J. Frokiaer1, and J. Leipziger2

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

* To whom correspondence should be addressed. E-mail: helle.praetorius{at}ki.au.dk.

The release of nucleotides is involved in mechano-sensation 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 if nucleotide release contributes to mechano-sensation in kidney epithelial cells, hereunder to establish whether pressure changes are sufficient to produce nucleotide mediated responses. 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 quantitydependent manner. The ATPase, apyrase, and the P2Y antgonist, suramin significantly reduced the pressure-induced Ca2+ transients. Applying apyrase or suramin to both sides of the preparation simultaneously almost abolished the pressure-induced Ca2+ response. In conclusion, these observations suggest that rapid pressure changes induce both apical and basolateral nucleotide release that contributes to mechano-sensation in kidney epithelial cells.




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