GABAA receptor beta subunit heterogeneity: functional expression of cloned cDNAs.

Abstract

Cloned cDNAs encoding two new beta subunits of the rat and bovine GABAA receptor have been isolated using a degenerate oligonucleotide probe based on a highly conserved peptide sequence in the second transmembrane domain of GABAA receptor subunits. The beta 2 and beta 3 subunits share approximately 72% sequence identity with the previously characterized beta 1 polypeptide. Northern analysis showed that both beta 2 and beta 3 mRNAs are more abundant in the brain than beta 1 mRNA. All three beta subunit encoding cDNAs were also identified in a library constructed from adrenal medulla RNA. Each beta subunit, when co‐expressed in Xenopus oocytes with an alpha subunit, forms functional GABAA receptors. These results, together with the known alpha subunit heterogeneity, suggest that a variety of related but functionally distinct GABAA receptor subtypes are generated by different subunit combinations.

Identification of Residues within GABAAReceptor α Subunits That Mediate Specific Assembly with Receptor β Subunits

“…8). These values are in accordance with a typical ␣1␤3 receptor profile (Yemer et al, 1989) and suggest that the mutation HY does not interfere per se with the ability of GABA to bind to the receptor and activate the ion channel. It therefore appears likely that the reduced responsiveness to GABA is a result of limited numbers of functional cell surface ␣␤ heteromers.…”

Autocrine/Paracrine Activation of the GABAAReceptor Inhibits the Proliferation of Neurogenic Polysialylated Neural Cell Adhesion Molecule-Positive (PSA-NCAM+) Precursor Cells from Postnatal Striatum

“…Cytosine arabinoside (10 M) was used as an internal control inhibiting proliferation in all phenotypes. Although agonists and antagonists of GABA A R did not modify the percentages of O4 ϩ /BrdU ϩ and GFAP ϩ /BrdU ϩ cells (data not shown), treatment with GABA A R agonists (GABA at 100 M and musci- (Khrestchatisky et al, 1989) GABA A R ␣2 For 5Ј-AGG TTG GTG CTG GCT AAC ATCC-3Ј Rev 5Ј-AAC AGA GTC AGA AGC ATT GTA AGT CC-3Ј 549 (Khrestchatisky et al, 1991) GABA A R ␣3 For 5Ј-CAA CAT AGT GGG AAC ACC TAT CC-3Ј Rev 5Ј-GGG AGC TCT GGG GTT TGG GAT TT-3Ј 350 (Criswell et al, 1997) GABA A R ␣4 For 5Ј-CAA AAC CTC CTC CAG AAG TTC CA-3Ј Rev 5Ј-ATG TTA AAT AAT GCC CCA AAT GTG ACT-3Ј 532 (Wisden et al, 1991) GABA A R ␣5 For 5Ј-TGA CCA AAA CCC TCC TTG TCT TCT-3Ј Rev 5Ј-ACC GCA GCC TTT CAT CTT TCC-3Ј 300 (Khrestchatisky et al, 1989) GABA A R ␤1 For 5Ј-GTT TGG GGC TTC TCT CTC TTT TCC T-3Ј Rev 5Ј-AGT TAC TGC TCC CTC TCC TCC ATT-3Ј 578 (Ymer et al, 1989a) GABA A R ␤2 For 5Ј-CAG GTT CTT ATC CCA GAT TGT CCC-3Ј Rev 5Ј-GGT CCA TCT TGT TGA CAT CCA GG-3Ј 408 (Ymer et al, 1989b) GABA A R ␤3 For 5Ј-CTT TTC GGC ATC TTC TCG GC-3Ј Rev 5Ј-TCC ACG CCA GTA ACA GCC TTG-3Ј 587 (Ymer et al, 1989b) GABA A R ␥1 For 5Ј-TAG TAA CAA TAA AGG AAA AAC CAC CAG A-3Ј Rev 5Ј-CCA GAT TGA ACA AGG CAA AAG CT-3Ј 296 GABA A R ␥2 For 5Ј-TGG TGA CTA TGT GGT TAT TGC CGT G-3Ј Rev 5Ј-AGG TGG GTG GCA TTG TTC ATT T-3Ј 423 (Khrestchatisky et al, 1989) GABA A R ␥3 For 5Ј-GAA ATC ATG GCG GCT CTA GTT-3Ј Rev 5Ј-CTC CAT CAG TGC GGC AAA GAC AAA-3Ј 336 (Stewart et al, 2002) GABA A R␦ For 5Ј-GAC TAC GTG GGC TCC AAC CTG GA-3Ј Rev 5Ј-ACT GTG GAG GTG ATG CGG ATG CT-3Ј 398 (Zhao and Joho, 1990) GAD 65 For 5Ј-CCA TTA CCC CAA TGA GCT TCT-3Ј Rev 5Ј-CCC CAA GCA GCA TCC ACG T-3Ј 698 (Dkhissi et al, 2001) GAD 67 For 5Ј-AAT TGC ACC CGT GTT TGT TCT TAT G-3Ј Rev 5Ј-AGC GCA GCC CCA GCC TTC TTT A-3Ј 252 (Stewart et al, 2002) For, Forward; Rev, reverse.…”

Identification, Molecular Cloning, and Characterization of a Novel GABAA Receptor-associated Protein, GRIF-1

“…The coding sequence of the human ␤2-IL (aa 301-426; Ref. 16) was amplified by the polymerase chain reaction (PCR) utilizing primers that contained EcoRI and BamHI restriction enzyme sites. The product was subcloned into the EcoRI and BamHI sites of the bait vector, pAS2-1 (Matchmaker Two-Hybrid System 2, CLONTECH, Palo Alto, CA).…”