The characterization and localization of the glutamate receptor subunit GluR1 in the rat brain.
The cloning of cDNAs that encode functional glutamate receptors makes it possible to produce antibodies that can be used as high-affinity probes for the localization and characterization of these receptors in the mammalian brain. We have made antibodies to different regions of the first cloned member of this family, GluR1, using bacterially overproduced antigen. On Western blots, these antisera detect glycoprotein(s) of 105 kDa present in crude membranes of the hippocampus and cerebellum. The 105-kDa band is associated with postsynaptic densities, and it is observed in cultured cells upon transfection with the GluR1 cDNA. Although glutamate receptors are thought to be the most prevalent excitatory ligand-gated ion channel in the mammalian brain, immunohistochemistry reveals that the receptors recognized by these antisera are localized predominantly in neurons of the cerebellum and some structures of the limbic system, including the hippocampus, the central nucleus of the amygdala, and portions of the septum. This pattern of expression is, in general, consistent with the distribution of GluR1 mRNA as determined by in situ hybridization histochemistry. Our results suggest that glutamate excitatory circuits recognized by these antisera are predominantly found in regions of the limbic system that are reciprocally interconnected.
Rogers
S W
SW
Molecular Neurobiology Laboratory, Salk Institute for Biological Studies, La Jolla, California 92037.
Hughes
T E
TE
Hollmann
M
M
Gasic
G P
GP
Deneris
E S
ES
Heinemann
S
S
D000818:Animals; D015153:Blotting, Western; D001923:Brain Chemistry; D002458:Cell Fractionation; D002460:Cell Line; D002462:Cell Membrane / Q000737:chemistry / Q000648:ultrastructure; D002531:Cerebellum / Q000737:chemistry; D004247:DNA / Q000235:genetics; D006031:Glycosylation; D006624:Hippocampus / Q000737:chemistry; D007150:Immunohistochemistry; D008970:Molecular Weight; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D011919:Rats, Inbred Strains; D017470:Receptors, Glutamate; D017981:Receptors, Neurotransmitter / Q000032:analysis* / Q000235:genetics; D013569:Synapses / Q000737:chemistry; D014018:Tissue Distribution; D014162:Transfection
D012333:RNA, Messenger; D017470:Receptors, Glutamate; D017981:Receptors, Neurotransmitter; D004247:DNA