Urea production by kidney collecting ducts in vitro: effect of amino acid addition.
Urea production by cortical (CCD) and medullary (OMCD) collecting ducts of the rat kidney was measured in vitro by incubating single microdissected pieces of tubule in the presence of L-[guanido-14C]arginine (0.2 mM). The [14C]urea released from the cells was hydrolysed in presence of urease added to the incubation medium and the 14CO2 formed was trapped in KOH and counted. The effect of various amino acids (AA) on urea production was investigated by adding unlabelled AA (either in combination or singly) at concentrations close to those present in blood plasma. A mixture of 17 AA decreased urea production from [14C]arginine by 46% in CCD and by 58% in OMCD. When lysine and proline were omitted from the mixture, the inhibition was less marked (19% in CCD and 43% in OMCD, respectively). When AA were tested singly, lysine induced the larger inhibition (40% in CCD and 45% in OMCD), than ornithine and glutamine (about 15% each, in CCD and OMCD), whereas proline inhibition (7% in CCD, 10% in OMCD) was not statistically significant. Branched-chain amino acids (BCAA) in combination (leucine, isoleucine and valine) also markedly reduced urea production by CCD and OMCD. Their effect was dose dependent. Solubilization of CCD and OMCD cell membranes with Triton X-100 resulted in a twofold increase in urea production by control samples; the relative inhibition (per cent) induced by BCAA was enhanced, whereas that induced by lysine was decreased. The data suggest that, in living tubules, the inhibition obtained with lysine resulted, for a large part, from competition between lysine and arginine for cell uptake via a common membrane carrier, whereas the inhibition induced by BCAA corresponded to an effect on arginase activity itself.
Levillain
O
O
Laboratoire de Physiologie Cellulaire, C.N.R.S. URA 219, Collège de France, Paris.
Hus-Citharel
A
A
Morel
F
F
D000596:Amino Acids / Q000378:metabolism / Q000494:pharmacology*; D000597:Amino Acids, Branched-Chain / Q000494:pharmacology; D000818:Animals; D001119:Arginase / Q000378:metabolism; D005973:Glutamine / Q000494:pharmacology; D066298:In Vitro Techniques; D007685:Kidney Tubules, Collecting / Q000378:metabolism*; D008297:Male; D009952:Ornithine / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D014508:Urea / Q000378:metabolism*
D000596:Amino Acids; D000597:Amino Acids, Branched-Chain; D005973:Glutamine; D014508:Urea; D009952:Ornithine; D001119:Arginase