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      In vitro Macro-and Microautoradiographic Localization of V 1 and V 2 Receptors in the Rat Kidney Using OPC-21268 and OPC-31260

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          Abstract

          To elucidate the precise localization of vasopressin (VP) Vj and V<sub>2</sub> receptors in the kidney, we utilized in vitro macroautoradiography (macro-ARG) and microautoradiography (micro-ARG) of these receptors in Wistar rat kidneys. This was done by using OPC-21268 and OPC-31260, two newly developed selective Vi (OPC-21268) and V<sub>2</sub> (OPC-31260) receptor antagonists. For macro-ARG, 10-μm kidney sections were incubated with Tris-HCl buffer containing [<sup>3</sup>H]-VP with or without unlabeled ligand (VP, OPC-21268, or OPC-31260) at 20°C for 40 min. These sections were then loaded into X-ray cassettes with Hyperfilm-[<sup>3</sup>H] and exposed in the dark for 2 months. The autoradiograms were quantitatively analyzed by using the research analysis system RAS 1,000; the V<sub>1</sub> and V<sub>2</sub> receptors were quantitated by subtracting the nonspecific binding (incubated with OPC-21268 and OPC-31260, respectively) from the total binding. To assess a more precise localization of the V<sub>1</sub>and V<sub>2</sub> receptors, we also investigated the micro-ARG of the renal V<sub>1</sub> and V<sub>2</sub> receptors by dipping the kidney section slides used for macro-ARG into a photographic emulsion and observing the receptors under light microscopy. [<sup>3</sup>H]-VP binding to the rat kidney was completely displaced by unlabeled excess VP, but not by unlabeled angiotensin II, indicating that [<sup>3</sup>H]-VP binding was specific for VP receptors. Computerized quantification showed that V<sub>2</sub> receptors, visualized by OPC-31260, were the predominant type of VP receptor in the kidney. Conversely, V<sub>1 </sub>receptors, visualized by OPC-21268, were fewer in number. V) receptors were partly localized to the glomerulus, cortical vessels, interstitial cells, and the medullary vessels. The V<sub>2 </sub>receptors localized to the collecting ducts and medullary tubules. Our findings indicated that renal V<sub>1</sub> and V<sub>2</sub> receptors can be detected by in vitro macro- and micro-ARG by using OPC-21268 and OPC-31260.

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          Author and article information

          Journal
          NEF
          Nephron
          10.1159/issn.1660-8151
          Nephron
          S. Karger AG
          1660-8151
          2235-3186
          1997
          1997
          23 December 2008
          : 76
          : 3
          : 331-336
          Affiliations
          a3rd Department of Internal Medicine, bHealth and Medical Center, and cDepartment of Neuroscience, Okayama University, Okayama, Japan
          Article
          190200 Nephron 1997;76:331–336
          10.1159/000190200
          9226235
          © 1997 S. Karger AG, Basel

          Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher. Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug. Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.

          Page count
          Pages: 6
          Categories
          Original Paper

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