Genes expressed in the brain define three distinct neuronal nicotinic acetylcholine receptors.

Nef, Patrick; Oneyser, Carole; Alliod, Christine; Couturier, Sabine; Ballivet, Marc

doi:10.1002/j.1460-2075.1988.tb02852.x

Cited by 227 publications

(82 citation statements)

References 57 publications

(24 reference statements)

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…To establish that the cerebellum contained the ␤2, ␤3, and ␤4 subunits as part of a receptor that binds acetylcholine in a nicotine-dependent manner, we performed immunodepletion experiments with antibodies against the three ␤ subunits and with an anti-␣4 subunit antibody. The ␣4 subunit was considered a likely partner in view of its general codistribution with the ␤2 subunit (Nef et al, 1988). Also, our transfection experiments indicated that the ␣4 subunit was the only ␣ subunit that could form a receptor with all three subunits (data not shown).…”

Section: Discussionmentioning

confidence: 79%

“…Because the ␣4 and ␤2 subunits are both broadly distributed in the mammalian brain and have been detected in Northern blots of chick cerebellum (Nef et al, 1988), we examined whether these two subunits were coassembled with the ␤3 and ␤4 subunits. Because the colocalization of the ␤3 and ␤4 subunits in rat cerebellum corresponds well with the immunocytochemical distribution of the ␤2 subunit (Hill, 1993), we used a mAb directed against the ␤2 subunit to immunoprecipitate receptor from detergent extracts of synaptosomes made from rat cerebellum.…”

Section: Coassembly Of the ␤3 And ␤4 Subunits In Cerebellummentioning

confidence: 99%

See 1 more Smart Citation

Formation of Oligomers Containing the β3 and β4 Subunits of the Rat Nicotinic Receptor

Forsayeth

Kobrin

1997

J. Neurosci.

View full text Add to dashboard Cite

The role of the ␤3 and ␤4 subunits of the nicotinic acetylcholine receptor in brain is still unclear. We investigated nicotinic receptor structure with antibodies directed against unique regions of the ␤3 and ␤4 subunits of the rat nicotinic acetylcholine receptor. Anti-␤4 detected a single band of 66 kDa in most regions of the brain that was strongest in striatum and cerebellum. The 60 kDa ␤3 subunit was detected primarily in striatum and cerebellum, and faintly in hippocampus. Immunoprecipitation experiments established that the two subunits were coassembled in the cerebellum along with the ␤2 subunit. Antibodies against the ␣4, ␤2, ␤3, and ␤4 subunits immunoprecipitated ϳ75% of the bungarotoxin-insensitive nicotinic receptor from cerebellar extracts as determined by nicotine-dependent acetylcholine binding. Transfection of COS cells with cDNAs for these four subunits induced expression of a high affinity nicotinic receptor. Omission of only a single subunit from the transfection affected either the B max or the apparent K D of the receptor. Our data suggest that the ␤3 subunit functions as a structural entity that links a relatively unstable ␣4␤2 heterodimer to a more stable ␣4␤4 heterodimer. The agonistbinding site formed by ␣4␤2 has a much greater affinity than does that formed by ␣4␤4. In this respect, nicotinic receptors that contain the ␤3 subunit are structurally homologous to the muscle nicotinic receptor.Key words: nicotine; acetylcholine; receptor; antibody; ␤3; ␤4; subunit Nicotinic acetylcholine receptors (nAChR) expressed in the CNS are members of a superfamily of ligand-gated ion channels that also include the muscle-type nAChR, GABA A , glycine, and 5-HT 3 receptors (Barnard et al., 1987;Egebjerg et al., 1991;Maricq et al., 1991;Moriyoshi et al., 1991;Monyer et al., 1992). Nine different rat neuronal nAChR subunit cDNAs (␣2, ␣3, ␣4, ␣5, ␣6, ␣7, ␤2, ␤3, and ␤4) have been cloned (for review, see Sargent, 1993). Most of the receptors in the family are thought to contain two or more different subunits, but the precise combinations of these subunits that exist in vivo are largely unknown. Previous work has indicated that the major high affinity nicotinic receptor in rat brain is composed of ␣4 and ␤2 subunits (Nakayama et al., 1991;Flores et al., 1992;Marks et al., 1992). More recent evidence indicates that deletion of the ␤2 subunit gene results in the loss of high affinity nicotine-binding sites from mouse brain (Picciotto et al., 1995). Nicotinic receptors have also been reconstituted in Xenopus oocytes. Thus, the ␤2 and ␤4 subunits can combine with various ␣ subunits to form functional receptors, permitting the formation of many types of receptor with unique pharmacological characteristics (Luetje and Patrick, 1991). Much less is known about the ␤3 subunit. It does not express any channel activity in oocytes in combination with any other single subunit , nor has it been demonstrated at the protein level in the CNS. Both the ␤3 and ␤4 subunits seem, by in situ hybridization, to have a more restric...

show abstract

Section: Discussionmentioning

confidence: 79%

Section: Coassembly Of the ␤3 And ␤4 Subunits In Cerebellummentioning

confidence: 99%

Formation of Oligomers Containing the β3 and β4 Subunits of the Rat Nicotinic Receptor

Forsayeth

Kobrin

1997

J. Neurosci.

View full text Add to dashboard Cite

show abstract

“…Northern blots were hybridized for 16 h at 42°C in 20% formamide, 6x SET, 10 x Denhardt's solution, 0.1% sodium pyrophosphate, 1% SDS, 100 kg/ml denatured herring sperm DNA and = lo6 cpm/ml radiolabeled probe. Final washes were in 1 x SET, 0.1 olo SDS at 55"C. , vertebrate neuronal genes [6,12], vertebrate muscle genes [6,29,30,32].…”

Section: Northern Analysismentioning

confidence: 99%

“…Within the highly conserved aminoterminal half comprising approximately amino acids l-330 [26] the sequence identity with ALS and ARD protein increases to 75% and 52'70, respectively. Homology values of this region between Da2 and subunits of vertebrate neuronal nAChRs range from =52% (for PZsubunits [5,6]) to =59% (for (~2-subunits [6,8]). The homology between Da2 and muscle receptor proteins is significantly lower.…”

Section: Structure Of the Dcx2 Proteinmentioning

confidence: 99%

Structure and developmental expression of the Dα2 gene encoding a novel nicotinic acetylcholine receptor protein of Drosophila melanogaster

et al. 1990

View full text Add to dashboard Cite

Nicotinic acetylcholine receptors (nAChRs) represent a heterogeneous group of excitatory neurotransmitter receptors in the insect brain. We have characterized the Da2 gene of Drosophila melanoguster, a new member of the nAChR gene family. The protein coding region is interrupted by six introns. The positions of three of these introns are shared with all other nAChR genes. The deduced Da2 protein shows the structural features of ligand-binding nAChR a-subunits. Cytogenetically, the Da2 gene maps at position 96A of the 3rd chromosome, close to the ALS gene which also encodes an a-like nAChR subunit. Da2 transcripts are predominantly expressed in late embryos and in fly heads.

show abstract

“…The α2 subunit was among the first non-muscle nAChR subunits to be cloned and heterologously expressed in a functional subtype, with the β2 subunit [3][4][5] . However, studies into α2 subunit containing (α2*) nAChRs were soon overtaken in number by those into other subtypes.…”

Section: Introductionmentioning

confidence: 99%

Pharmacological and immunochemical characterization of α2* nicotinic acetylcholine receptors (nAChRs) in mouse brain

et al. 2009

View full text Add to dashboard Cite

Aim: α2 nAChR subunit mRNA expression in mice is most intense in the olfactory bulbs and interpeduncular nucleus. We aimed to investigate the properties of α2* nAChRs in these mouse brain regions. Methods: α2 nAChR subunit-null mutant mice were engineered. Pharmacological and immunoprecipitation studies were used to determine the composition of α2 subunit-containing (α2*) nAChRs in these two regions. Results: [ 125 I]Epibatidine (200 pmol/L) autoradiography and saturation binding demonstrated that α2 deletion reduces nAChR expression in both olfactory bulbs and interpeduncular nucleus (by 4.8±1.7 and 92±26 fmol·mg -1 protein, respectively). Pharmacological characterization using the β2-selective drug A85380 to inhibit [ 125 I]epibatidine binding proved inconclusive, so immunoprecipitation methods were used to further characterize α2* nAChRs. Protocols were established to immunoprecipitate β2 and β4 nAChRs. Immunoprecipitation specificity was ascertained using tissue from β2-and β4-null mutant mice, and efficacy was good (>90% of β2* and >80% of β4* nAChRs were routinely recovered). Conclusion:Immunoprecipitation experiments indicated that interpeduncular nucleus α2* nAChRs predominantly contain β2 subunits, while those in olfactory bulbs contain mainly β4 subunits. In addition, the immunoprecipitation evidence indicated that both nuclei, but especially the interpeduncular nucleus, express nAChR complexes containing both β2 and β4 subunits.

show abstract

Genes expressed in the brain define three distinct neuronal nicotinic acetylcholine receptors.

Cited by 227 publications

References 57 publications

Formation of Oligomers Containing the β3 and β4 Subunits of the Rat Nicotinic Receptor

Formation of Oligomers Containing the β3 and β4 Subunits of the Rat Nicotinic Receptor

Structure and developmental expression of the Dα2 gene encoding a novel nicotinic acetylcholine receptor protein of Drosophila melanogaster

Pharmacological and immunochemical characterization of α2* nicotinic acetylcholine receptors (nAChRs) in mouse brain

Contact Info

Product

Resources

About