G. Pietig et al., Properties and regulation of organic cation transport in freshly isolated human proximal tubules, J BIOL CHEM, 276(36), 2001, pp. 33741-33746
The kidney, and more specifically the proximal tubule, is the main site of
elimination of cationic endogenous metabolites and xenobiotics. Although nu
merous studies exist on renal organic cation transport of rat and rabbit, n
o information is available from humans. Therefore, we examined organic cati
on transport and its regulation across the basolateral membrane of isolated
human proximal tubules. mRNA for the cation transporters hOCT1 and hOCT2 a
s well as hOCTN1 and hOCTN2 was detected in these tubules. Organic cation t
ransport across the basolateral membrane of isolated collapsed proximal tub
ules was recorded with the fluorescent dye 4-(4-dimethylamino)styryl-N-meth
ylpyridinium (ASP(+)). Depolarization of the cells by rising extracellular
K+ concentration to 145 mM reduced ASP(+) uptake by 20 +/- 5% (n = 15), ind
icating its electrogeneity. The substrates of organic cation transport tetr
aethylammonium (K-i = 63 muM) and cimetidine (K-i = 11 muM) as well as the
inhibitor quinine (K-i = 2.9 muM) reduced ASP(+) uptake concentration depen
dently. Maximal inhibition reached with these substances was similar to 60%
. Stimulation of protein kinase C with 1,2-dioctanoyl-sn-glycerol (DOG, 1 m
uM) or ATP (100 muM) inhibited ASP(+) uptake by 30 +/- 3 (n = 16) and 38 +/
- 13% (n = 6), respectively. The effect of DOG could be reduced with calpho
stin C (0.1 muM, n = 7). Activation of adenylate cyclase by forskolin (1 mu
M) decreased ASP(+) uptake by 29 +/- 3% (n = 10). hANP (10 nM) or 8-bromo-c
GMP (100 muM) also decreased ASP(+) uptake by 17 +/- 3 (n = 9) or 32 +/- 5%
(n = 10), respectively. We show for the first time that organic cation tra
nsport across the basolateral membrane of isolated human proximal tubules,
most likely mediated via hOCT2, is electrogenic and regulated by protein ki
nase C, the cAMP- and the cGMP-dependent protein kinases.