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Subapical Localization of the Dopamine D₃ Receptor in Proximal Tubules of the Rat Kidney

Asja Nürnberger, Marcus Räbiger, Andreas Mack, Jorge Diaz, Pierre Sokoloff, Bernd Mühlbauer and Gerd Luippold

Departments of Pharmacology and Toxicology (AN,MR,BM,GL) and Anatomy (AM), University of Tübingen, Tübingen, Germany; Unité de Neurobiologie et Pharmacologie Moléculaire (INSERM U 109), Centre Paul Broca (JD,PS), Paris, France; and Department of Clinical Pharmacology, Central Hospital Bremen (BM), Bremen, Germany

Correspondence to: PD Gerd Luippold, Dept. of Pharmacology and Toxicology, Faculty of Medicine, University of Tübingen, Wilhelmstr. 56, D-72074 Tübingen, Germany. E-mail: gerd.luippold{at}uni-tuebingen.de

The dopamine D₃ receptor (D₃R), intensively studied in neuroscience, also plays an important role in the regulation of renal and cardiovascular function. In contrast to functional findings, less information is available on its localization in the kidney. Neither RT-PCR studies nor radioligand binding assays are suitable to selectively determine the distribution of renal D₃R at the level of cellular or even subcellular structures. We studied the renal D₃R distribution in Sprague–Dawley rats by a polyclonal antiserum directed against an epitope in the third intracytoplasmic loop. D₃R immunoreactivity was detected by indirect immunofluorescence and confocal laser scanning microscopy. D₃R staining was confined to the renal cortex and occurred in proximal convoluted tubules near or in direct connection with the urinary pole of the glomeruli. The fluorescent spots were restricted to the subapical portion of the proximal tubular cells. Double staining with the F-actin marker phalloidin revealed a localization of the D₃R below the brush border region. However, staining by anti-ß₁/ß₂-adaptins, recognizing clathrin-coated compartments, did not correspond to the distribution of the D₃R signal. This is the first description of a D₃R accumulation in a cytoplasmic pool in the kidney, probably corresponding to a recycling mechanism or storage compartment. (J Histochem Cytochem 52:1647–1655, 2004)

Key Words: D₃ receptor • rat kidney • immunofluorescence • confocal laser scanning • microscopy • phalloidin • adaptin

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