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      • Record: found
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      • Article: found

      Changes of the Inducible Heat Shock Protein 70 mRNA Level in Rat Retina After Ischemia and Reperfusion

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          Abstract

          The heat shock protein 70 (HSP-70) mRNA level was evaluated in Long Evans rat retinas after ischemia and after reperfusion following ischemia. Retinal ischemia was induced by ligation of the optic nerve and vessels. Rats were sacrificed after 90 min of ischemia or 120 min of reperfusion following ischemia. Retinas were dissected. Total mRNA was extracted and inducible HSP-70 (iHSP-70) gene expression was analyzed by quantification of transcripts using an RT-PCR assay. Results were expressed in arbitrary units as a ratio of the optical density of iHSP-70/β-actin electrophoretic bands. iHSP-70 gene expression was 0.220 ± 0.027 (n = 5), 0.502 ± 0.045 (n = 5) and 0.468 ± 0.032 (n = 5) for the sham-operated, ischemia only and ischemia and reperfusion groups, respectively. There was a statistically significant difference between the control and ischemia groups, and between the control and ischemia and reperfusion groups (p < 0.001), suggesting a rapid HSP-70 mRNA expression of the retina due to an ischemic injury.

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          Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction.

          P. Chomczynski, N Sacchi (1987)
          A new method of total RNA isolation by a single extraction with an acid guanidinium thiocyanate-phenol-chloroform mixture is described. The method provides a pure preparation of undegraded RNA in high yield and can be completed within 4 h. It is particularly useful for processing large numbers of samples and for isolation of RNA from minute quantities of cells or tissue samples.
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            Quantitation of ischemic damage in the rat retina.

            W. F. Hughes (1991)
            In order to determine thresholds for irreversible cellular injury in the rat retina, timed acute no-flow ischemic episodes of 30-180 min duration were produced by elevation of intraocular pressure (IOP) above systolic pressure. Quantitation of irreversible degeneration and cell loss following a 2-week post-ischemic interval was performed by computer-assisted measurements from histologic sections. Alterations of thickness of retinal layers and linear cell density were determined for ischemia of selected durations (30, 60, 80, 90, 120 and 180 min). Different thresholds were evident for inner and outer retinal damage. Neurons of the inner nuclear layers showed extensive loss with episodes at 60 min. Decrease in the thickness of the inner plexiform layer provided the best index of this inner nuclear damage. The outer retina was more resistant, with photoreceptors showing extensive damage only after 90 min in conjunction with pigment epithelial metaplasia and degeneration. Two-hour episodes produced full-thickness degeneration with loss of pigment epithelium and sparing of the peripheral retina. Greater sensitivity of the inner retina suggested problems with restoration of the retinal circulation. Horseradish peroxidase infusions did reveal central microcirculatory defects in retinal wholemounts of some specimens with episodes longer than 60 min. Refinements of the methods resulted in outcomes sufficiently reproducible for quantitative assessment of acute ischemic injury. The rat retina provides an economical basic tissue model of acute ischemic injury affecting neurons, glia, and microvasculature. Quantitation of this injury promises great utility in testing agents with potentially protective effects on acute ischemic injury.
            Article 0 comments Cited 44 times – based on 0 reviews     Review now
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              Author and article information

              Journal
              Journal ID (publisher-id): ORE
              Journal ID (nlm-ta): Ophthalmic Res
              Journal ID (doi): 10.1159/issn.0030-3747
              Title: Ophthalmic Research
              Publisher: S. Karger AG
              ISSN (Print): 0030-3747
              ISSN (Electronic): 1423-0259
              Publication date Subscription-year: 1998
              Publication date (Print): October 1998
              Publication date (Electronic): 10 July 1998
              Volume: 30
              Issue: 5
              Pages: 291-294
              Affiliations
              a Department of Ophthalmology, University Hospital, and Laboratories of b Molecular Genetics and c Cardiovascular Physiology, University of Burgundy, Dijon, France
              Article
              Publisher ID: 55487 Other ID: Ophthalmic Res 1998;30:291–294
              DOI: 10.1159/000055487
              PubMed ID: 9704332
              SO-VID: 7856c076-b965-453e-9187-e1a764295597
              Copyright © © 1998 S. Karger AG, Basel
              License:

              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.

              History
              Page count
              Pages: 4
              Categories
              Subject: Original Paper

              ScienceOpen disciplines: Vision sciences,Ophthalmology & Optometry,Pathology
              Keywords: Retina,Ischemia-reperfusion,Heat shock protein
              Data availability:
              ScienceOpen disciplines: Vision sciences, Ophthalmology & Optometry, Pathology
              Keywords: Retina, Ischemia-reperfusion, Heat shock protein

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