Relative contribution of OAT and OCT transporters to organic electrolyte transport in rabbit proximal tubule.
We compared the characteristics of several cloned rabbit organic electrolyte (OE) transporters expressed in cultured cells with their behavior in intact rabbit renal proximal tubules (RPT) to determine the contribution of each to basolateral uptake of the weak acid ochratoxin A (OTA) and the weak base cimetidine (CIM). The activity of organic anion transporters OAT1 and OAT3 proved to be distinguishable because OAT1 had a high affinity for PAH (K(t) of 20 microM) and did not support estrone sulfate (ES) transport, whereas OAT3 had a high affinity for ES (K(t) of 4.5 microM) and a weak interaction with PAH (IC(50) > 1 mM). In contrast, both transporters robustly accumulated OTA. Intact RPT also accumulated OTA, with OAT1 and OAT3 each responsible for approximately 50%: ES and PAH each reduced uptake by approximately 50%, and the combination of the two eliminated mediated OTA uptake. The weak base CIM was transported by OAT3 (K(t) of 80 microM) and OCT2 (K(t) of 2 microM); OCT1 had a comparatively low affinity for CIM, and CIM uptake by OAT1 was equivocal. Intact RPT accumulated CIM, with TEA and ES reducing CIM uptake by 20 and 75%, respectively, suggesting that OAT3 plays a quantitatively more significant role in CIM uptake in the early proximal tubule than OCT1/2. In single S2 segments of RPT, ES and TEA each blocked approximately 50% of CIM uptake. Thus the fractional contribution of different OE transporters to renal secretion is influenced by their affinity for substrate and relative expression level in RPT.
Zhang
Xiaohong
X
Dept. of Physiology, College of Medicine, University of Arizona, Tucson, AZ 85724, USA.
Groves
Carlotta E
CE
Bahn
Andrew
A
Barendt
Wendy M
WM
Prado
Marcos D
MD
Rödiger
Matthias
M
Chatsudthipong
Varanuj
V
Burckhardt
Gerhard
G
Wright
Stephen H
SH
D000818:Animals; D002478:Cells, Cultured; D002927:Cimetidine / Q000378:metabolism; D003001:Cloning, Molecular; D003998:Dicarboxylic Acids / Q000378:metabolism; D004573:Electrolytes / Q000378:metabolism*; D004970:Estrone / Q000031:analogs & derivatives* / Q000494:pharmacology; D006635:Histamine H2 Antagonists / Q000378:metabolism; D007684:Kidney Tubules / Q000187:drug effects / Q000378:metabolism; D007687:Kidney Tubules, Proximal / Q000378:metabolism*; D007700:Kinetics; D009793:Ochratoxins / Q000494:pharmacology; D027362:Organic Anion Transport Protein 1 / Q000378:metabolism*; D029362:Organic Anion Transporters, Sodium-Independent / Q000378:metabolism*; D027702:Organic Cation Transporter 1 / Q000378:metabolism*; D012333:RNA, Messenger / Q000096:biosynthesis; D011817:Rabbits; D011994:Recombinant Proteins / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013379:Substrate Specificity; D014527:Uric Acid / Q000378:metabolism
D003998:Dicarboxylic Acids; D004573:Electrolytes; D006635:Histamine H2 Antagonists; D009793:Ochratoxins; D027362:Organic Anion Transport Protein 1; D029362:Organic Anion Transporters, Sodium-Independent; D027702:Organic Cation Transporter 1; D012333:RNA, Messenger; D011994:Recombinant Proteins; C114338:organic anion transport protein 3; C025589:ochratoxin A; D014527:Uric Acid; D004970:Estrone; D002927:Cimetidine; C017296:estrone sulfate