Neuroendocrinology
Original Paper
Colocalization of Peptide Histidine Isoleucine Amine and Corticotropin-Releasing Factor Immunoreactivity in Neurons of the Rat Hypothalamus: A Surprising ArtefactBerkenbosch F.a · Linton E.A.b · Tilders F.J.H.aaDepartment of Pharmacology, Medical Faculty, Free University, Amsterdam, The Netherlands; bDepartment of Chemical Pathology, St. Bartholomew’s Centre of Clinical Research, London, UK
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Article / Publication Details
Received: January 30, 1986
Accepted: June 11, 1986
Published online: April 01, 2008
Issue release date: 1986
Number of Print Pages: 9
Number of Figures: 0
Number of Tables: 0
ISSN: 0028-3835 (Print)
eISSN: 1423-0194 (Online)
For additional information: https://www.karger.com/NEN
Abstract
Indirect immunocytochemistry of corticotropin-releasing factor (CRF) and peptide histidine isoleucine amide (PHI) was performed by the use of antibodies raised to CRF and PHI. The staining intensity was quantitated by using an automated microfluorimeter. CRF and PHI immunoreactive fibres showed a similar pattern of distribution in the zona externa of the median eminence of the rat hypothalamus. Administration of colchicine (50 µg i.c.v.) resulted in the appearance of PHI and CRF immunoreactive cell bodies in the parvocellular part of the paraventricular nucleus. The PHI immunoreactive cell bodies were of low intensity and less abundant than those stained with the CRF antisera. Microfluorimetric measurements of the immunostaining in the median eminence showed parallel changes of PHI and CRF immunostaining after adrenalectomy, administration of reserpine and/or pargyline. In order to evaluate whether these data demonstrate that PHI and CRF are colocalized in hypothalamic neurons, we studied the specificity of PHI immunostaining by the use of a nonbiological gelatin model. Although CRF and PHI do not show structural homologies, the PHI antisera caused staining of PHI containing gels (range: 0.001–1 µM) but also of rat CRF (rCRF)-containing gels (range: 10–300 µM). In addition, preincubation of one of the PHI antisera with PHI or rCRF both caused a concentration-dependent quenching of the immunostaining in PHI- and CRF-containing gels and in preparations of the median eminence. Again, higher concentrations of rCRF (100 µM) than PHI (0.1 µM) were needed to show immunoinhibition, suggesting that the PHI antiserum has much lower avidity for native and fixed rCRF than for native and fixed PHI. We conclude that PHI immunostaining in the rat median eminence as found under the conditions used, is due, at least partially, to cross-reaction of the PHI antisera with rCRF.
© 1986 S. Karger AG, Basel
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Article / Publication Details
Received: January 30, 1986
Accepted: June 11, 1986
Published online: April 01, 2008
Issue release date: 1986
Number of Print Pages: 9
Number of Figures: 0
Number of Tables: 0
ISSN: 0028-3835 (Print)
eISSN: 1423-0194 (Online)
For additional information: https://www.karger.com/NEN
Copyright / Drug Dosage / Disclaimer
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.
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