A model for combined morphological and functional investigations on the isolated mediobasal rat hypothalamus

Details

Serval ID
serval:BIB_06A92CFE2B64
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
A model for combined morphological and functional investigations on the isolated mediobasal rat hypothalamus
Journal
Neuroendocrinology
Author(s)
Boujon  C. E., Bestetti  G. E., Reymond  M. J., Rossi  G. L.
ISSN
0028-3835
Publication state
Published
Issued date
04/1987
Peer-reviewed
Oui
Volume
45
Number
4
Pages
311-7
Notes
In Vitro
Journal Article
Research Support, Non-U.S. Gov't --- Old month value: Apr
Abstract
We have developed a model for combined morphological and functional in vitro studies of the isolated mediobasal hypothalamus (MBH) by considering two prerequisites: (1) the tissue must be well preserved, free of morphological artefacts and functionally unimpaired until the end of the in vitro incubation, and (2) the tissue must be processed for morphology in optimal conditions. To test our model we have studied some aspects of the luteinizing hormone-releasing hormone (LHRH) system in 4-month-old male Sprague-Dawley rats. After decapitation the MBH was isolated and put in a flask containing 0.5 ml Hepes-buffered Locke's medium gassed by 5 ml/min of O2/CO2 (95%/5%) and shaken in a water bath at 37 degrees C. After a 10-min washing, the medium was changed twice at an interval of 20 min. After the in vitro incubation the tissue was satisfactorily preserved as judged by light- and electron-microscopic analysis. LHRH, somatostatin and thyrotropin-releasing hormone could be demonstrated by alkaline phosphatase or peroxidase-antiperoxidase immunohistochemistry on semithin sections and by immunogold technique on thin sections. The LHRH secretion was close to basal values after 30 min of incubation (22.1 +/- 4.8 pg/MBH) and then remained constant for another period of 20 min (17.6 +/- 2.6 pg/MBH). During the second 20 min of incubation LHRH secretion increased in presence of 61.6 mM K+ (110.7 +/- 8.7 pg/MBH). Thus the isolated hypothalamus was excitable until the end of the in vitro incubation. We conclude that this model can be successfully used for combined morphological and functional studies.
Keywords
Animals Gonadotropin-Releasing Hormone/*metabolism Histocytochemistry Hypothalamus, Middle/analysis/*anatomy & histology/ultrastructure Immunoenzyme Techniques Male Microscopy, Electron Radioimmunoassay Rats Rats, Inbred Strains Somatostatin/analysis Thyrotropin-Releasing Hormone/analysis
Pubmed
Web of science
Create date
11/02/2008 14:11
Last modification date
20/08/2019 12:28
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