J. Bourke et al., UTP-preferring P-2 receptor mediates inhibition of sodium transport in porcine thyroid epithelial cells, BR J PHARM, 127(8), 1999, pp. 1787-1792
1 The effects of adenosine 5'-triphosphate (ATP). uridine 5'-triphosphate (
UTP) and analogues on forskolin-stimulated absorption of Na+ by porcine thy
roid epithelial cells were analysed in cultures grown as confluent monolaye
rs on permeable supports in Transwell Ussing chambers.
2 85% of the forskolin (10 mu M)-stimulated short-circuit current was inhib
ited by phenamil (1 mu M), which is a selective antagonist for epithelial t
ype Na+ channels.
3 Phenamil-sensitive current was inhibited in a dose dependent manner by nu
cleotides added to the apical compartment of Ussing chambers. In contrast,
the phenamil-resistant current, previously shown to represent anion secreti
on, was unaffected by nucleotides.
4 The order of potency (with EC50 values given in mu M) was UTP (0.08) > >
ATP (6.3) = uridine 5'-diphosphate (UDP) (6.6) > 2methyl-thio-adenosine-5'-
triphosphate (2MeSATP) (84.5) > adenosine 5'-diphosphate (ADP) (147.8) > al
pha,beta-methylene ATP (> 150) > >adenosine (> 1000).
5 P-2 receptors mediating inhibition of sodium absorption were present on t
he apical membrane of the cells since addition of UTP (1 - 1000 mu M) to th
e basal compartment of the Ussing chambers had little effect while subseque
nt addition to the apical compartment produced a normal response.
6 Cibachron blue (Reactive blue 2) (1-100 mu M), an antagonist at some P2 r
eceptor subtypes, inhibited phenamil sensitive current in a dose dependent
manner with half maximal inhibition occurring at 14.25 mu M.
7 Suramin (100 mu M), pyridoxalphosphate-6-azophenyl-2'.4'-disulphonic acid
(PPADS) (100 mu M) and pyridoxal 5'-phosphate (P5P) (100 mu M) showed only
slight competitive antagonism against the response to UTP.
8 These results indicate that a UTP-preferring P-2 receptor located on the
apical membrane of thyroid epithelial cells mediates inhibition of Na+ abso
rption.