Freeze-fracture immunocytochemical study of the expression of native and recombinant GABAA receptors.
To assess the density and distribution of native and recombinant GABAA receptors we used label-fracture and fracture-flip technologies combined with immunocytochemistry using monoclonal and polyclonal Abs directed against the extracellular domain of the GABAA receptor protein located in the freeze-fracture replicas. In cortical neurons there is a high density of GABAA receptors on both soma and dendrites with some areas were the density of receptors is higher, but there are no well defined clusters. In cerebellar granule cells most of the receptors are distributed in round clusters both in neurites and soma. In astroglial cells the receptor density is lower than in neurons and only occasionally they appear in clusters. In cells transfected with cDNAs encoding for various molecular forms of GABAA receptor subunits, the receptor density is moderate when cDNAs for alpha, beta and gamma subunits are cotransfected; however, on cells cotransfected with cDNAs for beta and gamma subunits the receptor density is significantly lower. Recombinant receptors appear randomly distributed and occasionally they aggregate in small groups.
Structural Biology Section, National Cancer Institute, Frederick Cancer Research and Development Center, MD 21702.
H J
HJ
Caruncho
G
G
Puia
E
E
Slobodyansky
P P
PP
da Silva
E
E
Costa
D000595:Amino Acid Sequence; D000818:Animals; D000831:Animals, Newborn; D000906:Antibodies; D001253:Astrocytes / Q000648:ultrastructure; D002460:Cell Line; D002478:Cells, Cultured; D002531:Cerebellum / Q000648:ultrastructure*; D002540:Cerebral Cortex / Q000648:ultrastructure*; D005614:Freeze Fracturing; D006801:Humans; D007668:Kidney; D046911:Macromolecular Substances; D016253:Microscopy, Immunoelectron; D008969:Molecular Sequence Data; D009474:Neurons / Q000648:ultrastructure*; D009842:Oligopeptides / Q000138:chemical synthesis / Q000276:immunology; D051381:Rats; D011963:Receptors, GABA-A / Q000032:analysis* / Q000378:metabolism; D011994:Recombinant Proteins / Q000032:analysis* / Q000378:metabolism; D014162:Transfection; D014407:Tumor Cells, Cultured
D000906:Antibodies; D046911:Macromolecular Substances; D009842:Oligopeptides; D011963:Receptors, GABA-A; D011994:Recombinant Proteins