Amino acid transport in podocytes.
It has recently been shown that formation of podocyte foot processes is dependent on a constant source of lipids and proteins (Simons M, Saffrich R, Reiser J, and Mundel P. J Am Soc Nephrol 10: 1633-1639, 1999). Here we characterize amino acid transport mechanisms in differentiated cultured podocytes and investigate whether it may be disturbed during podocyte injury. RT-PCR studies detected mRNA for transporters of neutral amino acids (ASCT1, ASCT2, and B(0/+)), cationic AA (CAT1 and CAT3), and anionic AA (EAAT2 and EAAT3). Alanine (Ala), asparagine, cysteine (Cys), glutamine (Gln), glycine (Gly), leucine (Leu), methionine (Met), phenylalanine (Phe), proline (Pro), serine (Ser), threonine (Thr), glutamic acid (Glu), arginine (Arg), and histidine (His) depolarized podocytes and increased their whole cell conductances. Depletion of extracellular Na(+) completely inhibited the depolarization induced by Ala, Gln, Glu, Gly, Leu, and Pro and decreased the depolarization induced by Arg and His, indicating the presence of Na(+)-dependent amino acid transport. Incubation of podocytes with 100 microg/ml puromycin aminonucleoside for 24 h significantly attenuated the effects induced by the various amino acids by approximately 70%. The data indicate the existence of different amino acid transporter systems in podocytes. Alteration of amino acid transport may participate in podocyte injury and disturbed foot process formation.
Gloy
J
J
Department of Medicine, Albert-Ludwigs-University Freiburg, Germany.
Reitinger
S
S
Fischer
K G
KG
Schreiber
R
R
Boucherot
A
A
Kunzelmann
K
K
Mundel
P
P
Pavenstädt
H
H
D000596:Amino Acids / Q000378:metabolism*; D000818:Animals; D001483:Base Sequence; D001693:Biological Transport, Active / Q000187:drug effects; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D002478:Cells, Cultured; D017931:DNA Primers / Q000235:genetics; D007668:Kidney / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D008564:Membrane Potentials; D051379:Mice; D011692:Puromycin Aminonucleoside / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012964:Sodium / Q000378:metabolism / Q000494:pharmacology
D000596:Amino Acids; D002352:Carrier Proteins; D017931:DNA Primers; D012333:RNA, Messenger; D011692:Puromycin Aminonucleoside; D012964:Sodium