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Arginine-vasopressin (AVP) stimulates vasopressin V2 receptors on the surface of renal collecting duct principal cells and thereby production of c AMP, which activates protein kinase A (PKA).Initiation of this signaling results in the redistribution of the water channel aquaporin-2 (AQP2) from intracellular vesicles into the plasma membrane by an exocytosis-like process.Aktories: Allgemeine und spezielle Pharmakologie und Toxikologie (11.

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The cells were fixed with paraformaldehyde, and AQP2 was detected by immunofluorescence microscopy using specific antibodies (H27) and Cy5-coupled antirabbit secondary antibodies (red).Here, we used a cell-based assay and automated immunofluorescence microscopy to screen 17,700 small molecules for inhibitors of the c AMP-dependent redistribution of aquaporin-2.This approach identified 17 inhibitors, including 4-acetyldiphyllin, a selective blocker of vacuolar H-ATPase that increases the p H of intracellular vesicles and causes accumulation of aquaporin-2 in the Golgi compartment.F-actin was visualized using TRITC-conjugated phalloidin (green).Fluorescence signals were detected using an automated screening microscope (10× magnification).

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