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Am J Physiol Renal Physiol 275: F285-F297, 1998;
0363-6127/98 $5.00
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Vol. 275, Issue 2, F285-F297, August 1998

Acute effects of vasopressin V2-receptor antagonist on kidney AQP2 expression and subcellular distribution

Birgitte Mønster Christensen1, David Marples4, Uffe Birk Jensen3, Jørgen Frøkiaer2, David Sheikh-Hamad5, Mark Knepper6, and Søren Nielsen1

1 Department of Cell Biology, Institute of Anatomy, 2 Department of Clinical Physiology, Aarhus University Hospital and Institute of Experimental Clinical Research, and 3 Institute of Human Genetics, University of Aarhus, DK-8000 Aarhus, Denmark; 4 Department of Physiology, University of Leeds; 5 Baylor College of Medicine, Department of Medicine, Nephrology Section, Houston, Texas 77030; and 6 Laboratory of Kidney and Electrolyte Metabolism, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892

The acute effect of treatment with the vasopressin V2-receptor antagonist OPC-31260 (OPC) on aquaporin-2 (AQP2) distribution and expression in rat kidney was examined. Immunofluorescence and semi-quantitative immunoelectron microscopy revealed that 15 and 30 min of OPC treatment resulted in significant reduction in apical plasma membrane labeling of AQP2, with a concomitant increase in labeling of vesicles and multivesicular bodies. In parallel, OPC treatment induced a large increase in urine output [0.6 ± 0.2 vs. 8.3 ± 1.0 ml/h (n = 4)]. Northern blotting using a 32P-labeled AQP2 cDNA probe and a digoxigenin-labeled AQP2 RNA probe revealed a band of ~1.6 kb corresponding to the predicted size of AQP2 mRNA. In control experiments, thirsting increased, whereas water loading decreased AQP2 mRNA levels. Treatment of rats with OPC caused a significant reduction in AQP2 mRNA within 30 min (52 ± 21%, n = 8, P < 0.025) and 60 min (56 ± 7%, n = 4, P < 0.001) of treatment compared with intravenous saline-injected controls. Thus a very rapid reduction in AQP2 mRNA was observed in response to vasopressin-receptor antagonist treatment. The reduction in AQP2 mRNA persisted after 24 h (40 ± 17%, n = 5, P < 0.05) of OPC treatment. There was a parallel increase in diuresis and reduction in urine osmolality. In conclusion, V2-receptor blockade produced a rapid internalization of AQP2 parallel with a rapid increase in urine output. Furthermore, OPC treatment caused a rapid and significant reduction in AQP2 mRNA expression, demonstrating that for rapid regulation of AQP2 expression, modulation of AQP2 mRNA levels is regulated via vasopressin-receptor signaling pathways.

aquaporin-2; collecting duct; nephrogenic diabetes insipidus; vasopressin antagonist


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