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Am J Physiol Renal Physiol (July 12, 2001). doi:10.1152/ajprenal.0349.2000
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Articles in PresS, published online ahead of print July 12, 2001
Am J Physiol Renal Physiol, 10.1152/ajprenal.0349.2000
Submitted on January 1, 2001
Accepted on June 14, 2001

ATP INHIBITS Mg2+ UPTAKE IN MOUSE DISTAL CONVOLUTED TUBULE CELLS VIA P2X PURINOCEPTORS

Long-Jun Dai1, Hyang Sub Kang1, Dirk Kerstan1, Gordon Ritchie1, and Gary A Quamme1*

1 Department of Medicine, University of British Columbia, Vancouver Hospital and Health Sciences Centre, Vancouver, BC, Canada

* To whom correspondence should be addressed. E-mail: QUAMME{at}unixg.ubc.ca.

Nucleotides have diverse effects on water and electrolyte reabsorption within the distal tubule of the nephron. As the distal tubule is important in control of renal magnesium balance, we determined the effects of ATP on cellular magnesium uptake in this segment. The effects of ATP were studied on immortalized mouse distal convoluted tubule (MDCT) cells by measuring Mg2+ uptake with fluorescence techniques. The mean basal Mg2+ uptake rate was 165±6 nM/s. ATP inhibited basal Mg2+ uptake and hormone-stimulated Mg2+ entry by 40 %. Both P2X (P2X1-P2X5 subtypes) and P2Y2 receptor subtypes were identified in MDCT cells using differential RT-PCR. Activation of both receptor subtypes with selective agonists increased intracellular Ca2+ concentration, P2X purinoceptors by ionotropic-gated channels and P2Y receptors via G protein-mediated intracellular Ca2+ release. The more relatively selective P2X agonists, ß, {gamma}-methylene ATP (ß, {gamma}-meATP) and benzoyl-benzoyl-ATP inhibited arginine vasopressin (AVP) and parathyroid hormone (PTH)-mediated Mg2+ uptake whereas agonists more selective for P2Y purinoceptors, UTP, ADP, and 2- methylthio ATP (2MeSATP) were without effect. Removal of extracellular Ca2+ diminished ß, {gamma}-meATP- mediated increase in intracellular Ca2+ and inhibition of AVP-stimulated Mg2+ entry. We conclude from this information that ATP inhibited Mg2+ uptake in MDCT cells through P2X purinoceptors expressed in this distal convoluted tubule cell line.




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