Podocyte-specific expression of organic cation transporter PMAT: implication in puromycin aminonucleoside nephrotoxicity.
Plasma membrane monoamine transporter (PMAT) is a novel polyspecific organic cation transporter that transports organic cations and the purine nucleoside, adenosine. PMAT is expressed in the kidney, but the specific localization and function of this transporter in renal cells are unclear. In this study, we developed a polyclonal antibody toward a 14-amino acid sequence in the last intracellular loop of PMAT and determined the precise cellular localization of PMAT in human and rat kidneys. Surprisingly, we found that the PMAT protein was predominantly expressed in the glomerulus with minimal expression in tubular cells. Within the glomerulus, dual-color immunofluorescence labeling showed that the PMAT protein was specifically localized to the visceral glomerular epithelial cells, i.e., podocytes. There was no significant PMAT immunoreactivity in mesangial or glomerular endothelial cells. We further showed that puromycin aminonucleoside (PAN), a classic podocyte toxin that induces massive proteinuria and severe glomerulopathy, is transported by PMAT. Expression of PMAT in Madin-Darby canine kidney cells significantly increased cell sensitivity to PAN. Decynium 22, a potent PMAT inhibitor, abolished PAN toxicity in PMAT-expressing cells. Together, our data suggest that PMAT is specifically expressed in podocytes and may play an important role in PAN-induced kidney injury.
Department of Pharmaceutics, H272J Health Sciences Bldg., Univ. of Washington, Seattle, WA 98195-7610, USA.
Li
L
Xia
Mingyan
M
Zhou
Thomas F
TF
Kalhorn
Horace T B
HT
Ho
Joanne
J
Wang
D000818:Animals; D000906:Antibodies / Q000276:immunology; D002460:Cell Line; D004285:Dogs; D004847:Epithelial Cells / Q000378:metabolism; D033705:Equilibrative Nucleoside Transport Proteins / Q000235:genetics / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000502:physiology*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007674:Kidney Diseases / Q000139:chemically induced*; D007678:Kidney Glomerulus / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D050199:Podocytes / Q000378:metabolism*; D011692:Puromycin Aminonucleoside / Q000378:metabolism / Q000633:toxicity*; D051381:Rats
D000906:Antibodies; D033705:Equilibrative Nucleoside Transport Proteins; C495988:SLC29A4 protein, human; D011692:Puromycin Aminonucleoside