Posts Tagged ‘P2Y purinoceptors’

ATP inhibits Mg2+ uptake in MDCT cells via P2X purinoceptors

Monday, September 14th, 2009

Dai LJ, Kand HS, Kerstan D, Ritchie G, and Quamme GA: ATP inhibits Mg2+ uptake in MDCT cells via P2X purinoceptorsAmerican Journal of Physiology 281:F833-F840, 2001.

PDF Download paper

Abstract 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 Mg21 balance, we determined the effects of ATP on cellular Mg21 uptake in this segment. The effects of ATP on immortalized mouse distal convoluted tubule (MDCT) cells were studied by measuring Mg21 uptake with fluorescence techniques. The mean basal Mg21 uptake rate was 165 6 6 nM/s. ATP inhibited basal Mg21 uptake and hormone-stimulated Mg21 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 Ca21 concentration, P2X purinoceptors by ionotropicgated channels, and P2Y receptors via G protein-mediated intracellular Ca21 release. The more relatively selective P2X agonists [b,g-methylene ATP (b,g-Me-ATP) and 29- and -39- O-(4-benzoyl-benzoyl)-ATP] inhibited arginine vasopressin (AVP)- and parathyroid hormone (PTH)-mediated Mg21 uptake whereas agonists more selective for P2Y purinoceptors (UTP, ADP, and 2-methylthio-ATP) were without effect. Removal of extracellular Ca21 diminished b,g-Me-ATP-mediated increase in intracellular Ca21 and inhibition of AVPstimulated Mg21 entry. We conclude from this information that ATP inhibited Mg21 uptake in MDCT cells through P2X purinoceptors expressed in this distal convoluted tubule cell line.

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 Mg21 balance, we determined
the effects of ATP on cellular Mg21 uptake in this segment.
The effects of ATP on immortalized mouse distal convoluted
tubule (MDCT) cells were studied by measuring Mg21 uptake
with fluorescence techniques. The mean basal Mg21 uptake
rate was 165 6 6 nM/s. ATP inhibited basal Mg21 uptake and
hormone-stimulated Mg21 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
Ca21 concentration, P2X purinoceptors by ionotropicgated
channels, and P2Y receptors via G protein-mediated
intracellular Ca21 release. The more relatively selective P2X
agonists [b,g-methylene ATP (b,g-Me-ATP) and 29- and -39-
O-(4-benzoyl-benzoyl)-ATP] inhibited arginine vasopressin
(AVP)- and parathyroid hormone (PTH)-mediated Mg21 uptake
whereas agonists more selective for P2Y purinoceptors
(UTP, ADP, and 2-methylthio-ATP) were without effect. Removal
of extracellular Ca21 diminished b,g-Me-ATP-mediated
increase in intracellular Ca21 and inhibition of AVPstimulated
Mg21 entry. We conclude from this information
that ATP inhibited Mg21 uptake in MDCT cells through P2X
purinoceptors expressed in this distal convoluted tubule cell
line.
intracellular magnesium, fluorescence; adenosine triphosphate