Laboratory Investigation

Kidney International (1981) 20, 772–779; doi:10.1038/ki.1981.210

Vanadate inhibits urinary acidification by the turtle bladder

Jose A L Arruda1, Sandra Sabatini1, Christof Westenfelder1, With technical assistance of George Dytko1, Gabriel Achanzar1, Jell Hsieh1, Elizabeth M Sack1 and Wayne R Earnest1

1Sections of Nephrology, University of Illinois Abraham Lincoln School of Medicine and the Veterans Administration West Side Hospital, Chicago, Illinois

Correspondence: Dr J A L Arruda, Department of Medicine, Division of Nephrology, University of Illinois Hospital, 840 South Wood Street, Chicago, Illinois 60612, USA

Received 29 December 1980; Revised 7 April 1981.

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Abstract

Vanadate inhibits urinary acidification by the turtle bladder. We studied the effect of vanadate on urinary acidification by the turtle bladder in vitro. Vanadate added to the serosal, but not to the mucosal, solution caused a dose dependent inhibition of hydrogen ion secretion in short circuited hemibladders in the presence and in the absence of ouabain. The failure of mucosal addition of vanadate to inhibit hydrogen ion secretion was due to the fact that vanadate tissue uptake from the serosal solution was 30 times greater than that from the mucosal solution, suggesting that vanadate must enter lhe cell to inhibit hydrogen ion secretion. The effect of vanadate on hydrogen ion secretion could not be explained by alterations in calcium uptake or efflux. The effect of vanadate on hydrogen ion secretion under aerobic conditions could not be explained solely by inhibition of mitochondrial function because the relative magnitude of the inhibitory effect of this compound was the same under aerobic and anaerobic conditions. Vanadate caused a significant inhibition of the proton motive force (PMF) without altering backleak of hydrogen ion or bicarbonate secretion. Under anaerobic conditions vanadate inhibited hydrogen ion secretion and caused a significant increase in lactate production without the expected increase in ATP/ADP ratio. ATP levels under anaerobic conditions were not significantly different between control and vanadate treated hemibladders, indicating that limited ATP availability can not explain the inhibitory effect of vanadate on hydrogen ion secretion. These findings indicate that vanadate has a number of effects that are capable of explaining the inhibition of urinary acidification. Among those effects, the inhibition of hydrogen ion secretion and the PMF by vanadate may be due to direct inhibition of the hydrogen ion pump, interference with ATP utilization by the pump, or other metabolic effects.

Le vanadate inhibe l'acidification urinaire par la vessie de tortue. Nous avons étudié in vitro l'effet du vanadate sur l'acidification urinaire par la vessie de tortue. La présence de vanadate dans le bain séreux, mais non dans le bain muqueux, détermine une inhibition, dépendante de la dose, de la sécrétion d'ions d'hydrogène dans les hémi-vessies en court-circuit, en présence et en l'absence d'ouabaïne. L'absence d'effet de l'addition de vanadate à la face muqueuse est due au fait que la captation tissulaire de vanadate est 30 fois plus grande à partir du bain séreux que du bain muqueux, ce qui suggère que le vanadate doit pénétrer dans la cellule pour inhiber la sécrétion d'ion hydrogène. L'effet du vanadate sur la sécrétion d'ions d'hydrogène ne peut être expliqué par les modifications de la captation ou de l'efflux de calcium. L'effet du vanadate sur la sécrétion d'ions d'hydrogène dans des conditions aérobies ne peut être expliqué seulement pas l'inhibition de la fonction mitochondriale puisque l'importance relative de l'effet inhibiteur est la même dans les conditions aérobies et anaérobies. Le vanadate détermine une inhibition significative de la force motrice des protons (PMF) sans modification de la rétrodiffusion d'ions d'hydrogène ou de la sécrétion de bicarbonate. En condition anaérobie, le vanadate inhibe la sécrétion d'ions d'hydrogène et détermine une augmentation significative de la production de lactate sans l'augmentation attendue du rapport ATP/ADP. Les concentrations d'ATP en condition anaérobie ne sont pas significativement différentes entre les hémi-vessies contrôles et traitées par le vanadate, ce qui indique que la disponibilité d'ATP ne peut expliquer l'effet inhibiteur du vanadate sur la sécrétion d'ions d'hydrogène. Ces constatations indiquent que le vanadate a de nombreux effets qui sont capables de participer à l'inhibition de l'acidification urinaire. Parmi ces effets l'inhibition de la sécrétion d'ions d'hydrogène et de la PMF par le vanadate peut être due à une inhibition directe de la pompe ions d'hydrogène, à une interférence avec l'utilisation d'ATP par la pompe ou d'autres effets métaboliques.

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