Expression of OCTN2 and OCTN3 in the apical membrane of rat renal cortex and medulla.
Immunological assays and transport measurements in apical membrane vesicles revealed that the apical membrane of rat kidney cortex and medulla presents OCTN2 and OCTN3 proteins and transports L-[(3)H]-carnitine in a Na(+)-dependent and -independent manner. OCTN2 mediates the Na(+)/L-carnitine transport activity measured in medulla because (i) the transport showed the same characteristics as the cortical Na(+)/L-carnitine transporter and (ii) the medulla expressed OCTN2 mRNA and protein. The Na(+)-independent L-carnitine transport activity appears to be mediated by both OCTN2 and OCTN3 since: (i) Na(+)-independent L-carnitine uptake was inhibited by both, anti-OCTN2 and anti-OCTN3 antibodies, (ii) kinetics studies revealed the involvement of a high- and a low-affinity transport systems, and (iii) Western and immunohistochemistry studies revealed that OCTN3 protein is located at the apical membrane of the kidney epithelia. The Na(+)-independent L-carnitine uptake exhibited trans-stimulation by intravesicular L-carnitine or betaine. This trans-stimulation was inhibited by anti-OCTN3 antibody, but not by anti-OCTN2 antibody, indicating that OCTN3 can function as an L-carnitine/organic compound exchanger. This is the first report showing a functional apical OCTN2 in the renal medulla and a functional apical OCTN3 in both renal cortex and medulla.
Cano
M M
MM
Facultad de Farmacia, Departamento de Fisiología y Zoología (Biomembranes Group), Universidad de Sevilla, 41012Sevilla, Spain.
Calonge
M L
ML
Ilundain
A A
AA
D000818:Animals; D019138:Antibodies, Blocking / Q000494:pharmacology; D001622:Betaine / Q000378:metabolism; D001693:Biological Transport, Active / Q000502:physiology; D002331:Carnitine / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism* / Q000648:ultrastructure; D004847:Epithelial Cells / Q000378:metabolism* / Q000648:ultrastructure; D007150:Immunohistochemistry; D007684:Kidney Tubules / Q000378:metabolism* / Q000648:ultrastructure; D008297:Male; D051379:Mice; D027701:Organic Cation Transport Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D012964:Sodium / Q000378:metabolism; D000074058:Solute Carrier Family 22 Member 5; D022161:Transport Vesicles / Q000378:metabolism / Q000648:ultrastructure
D019138:Antibodies, Blocking; D027701:Organic Cation Transport Proteins; D012333:RNA, Messenger; C498687:Slc22a21 protein, mouse; C115441:Slc22a5 protein, rat; D000074058:Solute Carrier Family 22 Member 5; D001622:Betaine; D012964:Sodium; D002331:Carnitine