Localization of GABA(B) (R1) receptors in the rat hippocampus by immunocytochemistry and high resolution autoradiography, with specific reference to its localization in identified hippocampal interneuron subpopulations.
Immunocytochemical and autoradiographic methods were used to localize the GABA(B) receptor in the normal rat hippocampus. GABA(B) receptor 1-like immunoreactivity (GBR1-LI) was most intense in presumed GABAergic interneurons of all hippocampal subregions. It was also present throughout the hippocampal neuropil, where it was most intense in the dendritic strata of the dentate gyrus, which are innervated by the perforant pathway and inhibitory dentate hilar cells, and in strata oriens and radiatum of area CA3. The dendritic regions of area CA1 exhibited less GBR1-LI than area CA3. GBR1-LI was detectable in the somata of CA1 pyramidal cells, but was minimal or undetectable within the somata of dentate granule cells and CA3 pyramidal cells. GBR1-LI was similarly minimal in the dentate hilar neuropil, and in stratum lucidum, the two regions that contain granule cell axons and terminals. Nor was GBR1-LI detectable in the inhibitory basket cell fiber systems that surround hippocampal principal cell somata. Fluorescence co-localization studies indicated that significant proportions of interneurons expressing somatostatin, neuropeptide Y, cholecystokinin, calbindin, or calretinin also expressed GBR1-LI constitutively. Conversely, parvalbumin-positive GABAergic basket cells of the dentate gyrus and hippocampus, which form GABA(A) receptor-mediated inhibitory axo-somatic synapses, rarely contained detectable GBR1-LI. High resolution autoradiography with the GABA(B) receptor antagonist CGP 62349 revealed a close correspondence between receptor ligand binding and GBR1-LI, with several notable exceptions. Ligand binding closely matched GBR1-LI throughout the hippocampal, cortical, thalamic, and cerebellar neuropil. However, the hippocampal interneuron somata and dendrites that exhibited the most intense GBR1-LI, and the GBR1-positive somata of CA1 pyramidal cells, did not exhibit a similar density of [3H]-CGP 62349 binding. These data clarify the relationship between immunocytochemically identified receptor protein and potentially functional receptors, indicating that GBR1-LI reflects both non-functional cytoplasmic GBR1 and the ligand-bindable form of the protein, both before dimerization with GBR2 and after translocation to functional sites within cells. The staining and binding patterns further suggest that GBR1 is constitutively expressed in specific neuronal populations, and may exist in higher concentration in the axons of inhibitory hippocampal pathways that innervate dendritic zones, than in axo-somatic inhibitory terminals. Whether GBR1 is inducible in cells that contain GBR1 mRNA, but no detectable constitutive protein, remains to be determined in experimental studies.
Sloviter
R S
RS
Department of Pharmacology, University of Arizona College of Medicine, Tucson 85724-5050, USA. sloviter@u.arizona.edu
Ali-Akbarian
L
L
Elliott
R C
RC
Bowery
B J
BJ
Bowery
N G
NG
D000818:Animals; D001345:Autoradiography; D001565:Benzoates / Q000494:pharmacology; D018756:GABA Antagonists / Q000494:pharmacology; D006624:Hippocampus / Q000737:chemistry* / Q000187:drug effects; D007150:Immunohistochemistry; D007395:Interneurons / Q000737:chemistry* / Q000187:drug effects; D008297:Male; D009943:Organophosphorus Compounds / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D018079:Receptors, GABA / Q000032:analysis* / Q000187:drug effects; D011963:Receptors, GABA-A; D018080:Receptors, GABA-B
D001565:Benzoates; C114540:CGP 62349; D018756:GABA Antagonists; C491397:Gabrr2 protein, rat; D009943:Organophosphorus Compounds; D018079:Receptors, GABA; D011963:Receptors, GABA-A; D018080:Receptors, GABA-B