Acute effect of cadmium-metallothionein on glucose and amino acid transport across the apical membrane of the rabbit proximal tubule perfused in vitro.
Acute as well as chronic exposure of cadmium (Cd) leads to proximal tubule injury. The exact cellular mechanism of this disorder and whether there is a contribution of cadmium-metallothionein (Cd-MT), a binding protein of Cd, remain unclear. We perfused isolated S2 segments of rabbit nephron, and the deflections of transmural voltage (DeltaV(t)) and apical membrane voltage (DeltaV(a)) on elimination of glucose or alanine from the perfusate were measured for the parameters of activity of Na(+)-glucose and Na(+)-amino acid cotransporters. The effects of Cd-MT or CdCl(2) to either bath or lumen for 10 min on these parameters were examined. We also measured the lumen-to-bath [(14)C]glucose flux. Addition of Cd-MT to lumen suppressed glucose- or alanine-dependent DeltaV(t) and DeltaV(a), as well as baseline V(t) and basolateral membrane voltage (V(b)), at approximately 10 min. [(14)C]glucose flux was inhibited by Cd-MT to lumen. The effects of Cd-MT to bath and CdCl(2) to either lumen or bath were 100-fold less potent than that of Cd-MT to lumen. Luminal Cd-MT immediately suppressed the glucose-dependent DeltaV(a), whereas the baseline V(a) and V(t) were unchanged. The early effect of luminal Cd-MT was simulated by addition of 10(-4) M phloretin. Addition of 10(-4) M ouabain to the bath simulated the later effect of Cd-MT. The protection of SH group by dithiothreitol prevented the early effect of Cd-MT, but not the later effect. We concluded that Cd-MT initially acts directly on Na(+)-glucose and Na(+)-amino acid cotransporters from the lumen by attacking SH group, followed by the later inhibition of Na(+)-K(+)-ATPase after entering the cell from the apical membrane.
Tsuruoka
S
S
Department of Clinical Pharmacology, Jichi Medical School, Tochigi, Japan. tsuru@jichi.ac.jp
Sugimoto
K
K
Muto
S
S
Nomiyama
K
K
Fujimura
A
A
Imai
M
M
D000409:Alanine / Q000378:metabolism*; D000818:Animals; D002104:Cadmium / Q000494:pharmacology*; D019256:Cadmium Chloride / Q000378:metabolism; D004229:Dithiothreitol / Q000494:pharmacology; D004347:Drug Interactions; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005947:Glucose / Q000378:metabolism*; D066298:In Vitro Techniques; D007687:Kidney Tubules, Proximal / Q000187:drug effects* / Q000378:metabolism; D008564:Membrane Potentials / Q000187:drug effects; D008668:Metallothionein / Q000378:metabolism*; D009399:Nephrons / Q000187:drug effects / Q000378:metabolism; D010042:Ouabain / Q000494:pharmacology; D010477:Perfusion; D010693:Phloretin / Q000494:pharmacology; D011485:Protein Binding; D011817:Rabbits; D012964:Sodium / Q000502:physiology; D013997:Time Factors
D004791:Enzyme Inhibitors; D002104:Cadmium; D010042:Ouabain; D008668:Metallothionein; D012964:Sodium; D005947:Glucose; D019256:Cadmium Chloride; D000409:Alanine; D010693:Phloretin; D004229:Dithiothreitol