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Institute of Physiology, Albert-Ludwigs-University, Freiburg, Germany.
Correspondence to Dr. Jens Leipziger, Physiologisches Institut, Albert-Lud-wigs-Universität, Hermann-Herder-Straße 7, 79104 Freiburg, Germany. Fax: 0761-203-5191; E-mail: leipzige{at}ruf.uni-freiburg.de
Abstract. Extracellular nucleotides regulate renal ion transport.
With the use of in vitro perfusion and [Ca2+]i
imaging, this study investigated whether mouse and rabbit cortical collecting
ducts (CCD) respond to luminal nucleotides. In mouse CCD, luminal ATP
(EC50: 10 µM) and UTP (EC50: 9.7 µM) increased
[Ca2+]i with an initial peak and a plateau. To make
certain that basolateral P2 receptors were not activated by luminal
nucleotides via leak diffusion, luminal trypsin (1 µM), a known agonist for
basolateral proteinase-activated receptors, was perfused. Mouse CCD that were
responsive to luminal ATP were nonresponsive to luminal trypsin but always
showed [Ca2+]i elevations by basolateral trypsin (10 or
100 nM). Luminal 
,ß- and ß,
-methylene ATP,
2-methyl-S-ATP, ADP, UDP, and 2',3'-O-4-benzoylbenzoyl ATP had no
effect (100 µM, n = 9). Without external Ca2+, luminal
ATP still stimulated a [Ca2+]i increase. Mouse CCD also
responded to basolateral ATP (EC50: 23 µM) and UTP
(EC50: 23 µM) with smaller [Ca2+]i
elevations. Confocal microscopy of perfused CCD showed that luminal ATP (100
µM) rapidly increased [Ca2+]i in nearly all cells
(n = 6) and the same cells that responded to luminal ATP responded to
basolateral ATP (100 µM). In contrast, rabbit CCD did not respond to
luminal ATP/UTP (n = 8) despite ATP's known effect from the
basolateral side (EC50: 34 µM). These data indicate the
expression of luminal P2Y receptors (probably P2Y2) in principal
cells of mouse CCD but not in rabbit CCD.
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