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Immunohistochemical localization of a GHB receptor‐like protein isolated from rat brain
Author(s) -
Kemmel Véronique,
Miehe Monique,
Roussel Guy,
Taleb Omar,
NailBoucherie Karine,
Marchand Christiane,
Stutz Christine,
Andriamampandry Christian,
Aunis Dominique,
Maitre Michel
Publication year - 2006
Publication title -
journal of comparative neurology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.855
H-Index - 209
eISSN - 1096-9861
pISSN - 0021-9967
DOI - 10.1002/cne.21072
Subject(s) - biology , receptor , colocalization , neurotransmitter receptor , microbiology and biotechnology , compartmentalization (fire protection) , postsynaptic potential , neurotransmitter , polyclonal antibodies , biochemistry , neuroscience , antibody , enzyme , immunology
Gamma‐hydroxybutyrate (GHB) is a substance derived from the metabolism of GABA and is heterogeneously distributed in various regions of the brain. This compound possesses a neuromodulatory role on several types of synapses, particularly those using GABA as a neurotransmitter. At physiological concentrations, this effect of GHB is mediated via specific receptors that induce neuronal hyperpolarization and bind radioactive GHB with a specific distribution, ontogenesis, kinetics, and pharmacology. A membrane protein that possesses six to seven transmembrane domains and which binds and is activated by micromolar amounts of GHB was recently cloned from rat brain hippocampus. In order to study the regional and cellular distribution of this receptor in rat brain, we selected several specific peptides belonging to the extracellular domains of the receptor to be used as specific immunogens to raise polyclonal antibodies in the rabbit. Among the antisera obtained, one of them gave particularly good results in terms of specificity and reactivity at high dilution. Immunohistochemical analyses, both at the confocal and electron microscopic level, showed receptor protein distribution closely resembling the distribution of GHB high‐affinity binding sites, except for cerebellum, where GHB receptor(s) of lower affinity exist(s). In all regions studied the GHB receptor‐like protein labeling appears to be distributed specifically in neurons and not in glial cells. At the cellular level the antibody specifically labels dendrites, and no immunoreactivity was detected in presynaptic endings or in axons. Accordingly, electron microscopy reveals strong labeling of postsynaptic densities and of neuronal cytosol. J. Comp. Neurol. 498:508–524, 2006. © 2006 Wiley‐Liss, Inc.