Exon–Intron Structure of the Human Neuronal Nicotinic Acetylcholine Receptor α4 Subunit (CHRNA4)

Steinlein, Ortrud K.; Weiland, Sigrid; Stoodt, Jens; Propping, Peter

doi:10.1006/geno.1996.0119

Cited by 48 publications

(24 citation statements)

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“…cDNA encoding a human ␣4 subunit (Steinlein et al, 1996; see GenBank accession NM_000744 for an update) was excised from the pSPoD vector as a HindIII-SalI fragment encompassing 54 bp of sequences in the vector polycloning site in the 5Ј direction from the 1937 bp of ␣4 cDNA and 204 bp corresponding to the vector polycloning site and some vector sequences in the 3Ј direction from the insert. The ends of the fragment were blunt-ended using Klenow enzyme and ligated into pcDNA3.1/zeo (Invitrogen) cut at the EcoRV site to generate the pcDNA3.1/zeo-h␣4 construct.…”

Section: Methodsmentioning

confidence: 99%

Characterization of Human α4β2-Nicotinic Acetylcholine Receptors Stably and Heterologously Expressed in Native Nicotinic Receptor-Null SH-EP1 Human Epithelial Cells

et al. 2003

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Naturally expressed nicotinic acetylcholine receptors composed of ␣4 and ␤2 subunits (␣4␤2-nAChR) are the predominant form of high affinity nicotine binding site in the brain implicated in nicotine reward, mediation of nicotinic cholinergic transmission, modulation of signaling through other chemical messages, and a number of neuropsychiatric disorders. To develop a model system for studies of human ␣4␤2-nAChR allowing protein chemical, functional, pharmacological, and regulation of expression studies, human ␣4 and ␤2 subunits were stably introduced into the native nAChR-null human epithelial cell line SH-EP1. Heterologously expressed ␣4␤2-nAChR engage in high-affinity, specific binding of Rbϩ efflux assays indicate full efficacy of epibatidine, nicotine, and acetylcholine; partial efficacy for 1,1-dimethyl-4-phenyl-piperazinium, cytisine, and suberyldicholine; competitive antagonism by dihydro-␤-erythroidine, decamethonium, and methyllycaconitine; noncompetitive antagonism by mecamylamine and eserine; and mixed antagonism by pancuronium, hexamethonium, and d-tubocurarine. These results demonstrate utility of transfected SH-EP1 cells as models for studies of human ␣4␤2-nAChR, and they also reveal complex relationships between apparent affinities of drugs for radioligand binding and functional sites on human ␣4␤2-nAChR.

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Section: Methodsmentioning

confidence: 99%

Characterization of Human α4β2-Nicotinic Acetylcholine Receptors Stably and Heterologously Expressed in Native Nicotinic Receptor-Null SH-EP1 Human Epithelial Cells

et al. 2003

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“…Only the structure of CHRNA3 was previously reported, although its exact genomic size had not been evaluated (Rempel et al 1998). The exon-intron structures of CHRNA5 and CHRNB4 were identified and shown to be characterized by the presence of six exons and five introns, as observed for other nAChR coding genes (e.g., CHRNA4 and CHRNB2) (Steinlein et al 1996;Rempel et al 1998). Moreover, partial sequences of the 5Ј and 3Ј ends of each intron were determined for both genes.…”

Section: Discussionmentioning

confidence: 75%

“…Although cDNA sequences for nAChR subunits have been analyzed in humans and in a number of different species, the genomic structure has been described only for CHRNA3, CHRNA4, CHRNA7, CHRNA10, and CHRNB2, coding for the α3, α4, α7, α10, and 2 subunits, respectively (Steinlein et al 1996;Gault et al 1998;Rempel et al 1998;Lustig et al 2001). The overall genomic organization of these genes is conserved and characterized by the presence of six exons, except for CHRNA7, which, both in humans and chick, consists of ten exons as the result of partial gene duplication.…”

Section: Introductionmentioning

confidence: 99%

Characterization of the genomic structure of the human neuronal nicotinic acetylcholine receptor CHRNA5/A3/B4 gene cluster and identification of novel intragenic polymorphisms

et al. 2001

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Genes coding for the α5, α3, and 4 subunits (CHRNA5, CHRNA3, and CHRNB4) of the neuronal nicotinic acetylcholine receptors (nAChRs) are clustered on chromosome 15q24. Linkage of this chromosomal region to autosomal dominant nocturnal frontal lobe epilepsy (ADNFLE), an idiopathic partial epilepsy, was reported in one family. Moreover, mutations in other neuronal nAChR subunit genes coding for the α4 (CHRNA4) and the 2 (CHRNB2) subunits were associated with ADNFLE. Apart from the exon-intron structure of CHRNA3, the genomic organization of this gene cluster was unknown, making comprehensive mutational analyses impossible. The genomic structure of CHRNA5 and CHRNB4 is here reported. Moreover, two hitherto unknown introns were identified within the 3Ј untranslated region of CHRNA3, causing a partial tail-to-tail overlap with CHRNA5. Four novel intragenic polymorphisms were identified and characterized in the cluster.

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“…Among others, chromosome 20p11.21 harbors genes coding for Type 2 cystatins (CSTs), extracellular secreted polypeptides that are broadly distributed and found in most body fluids (Dickinson, Thiesse, & Hicks, 2002). Interestingly, there is a resemblance between cystatins and the a subunits of nAChRs, as both contain four cysteine residues forming two disulfide bonds, which in nAChRs play a critical role in agonist binding (Steinlein, Weiland, Stoodt, & Propping, 1996). Chromosome 20p11.21 also contains GGTLC1 (gamma-glutamyltransferase light chain), a transpeptidase crucial in the metabolism of glutathione.…”

Section: Discussionmentioning

confidence: 99%

Chromosome 20 Shows Linkage With DSM-IV Nicotine Dependence in Finnish Adult Smokers

Keskitalo-Vuokko

Hällfors

Broms

et al. 2011

Nicotine & Tobacco Research

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Introduction: Chromosome 20 has previously been associated with nicotine dependence (ND) and smoking cessation. Our aim was to replicate and extend these findings.Methods: First, a total of 759 subjects belonging to 206 Finnish families were genotyped with 18 microsatellite markers residing on chromosome 20, in order to replicate previous linkage findings. Then, the replication data were combined to an existing wholegenome linkage data resulting in a total of 1,302 genotyped subjects from 357 families. ND diagnosed by DSM-IV criteria, the Fagerström Test for Nicotine Dependence (FTND) score, and the lifetime maximum number of cigarettes smoked within a 24-hr period (MaxCigs24) were used as phenotypes in the nonparametric linkage analyses. Results:We replicated previously reported linkage to DSM-IV ND, with a maximum logarithm of odd (LOD) score of 3.8 on 20p11, with females contributing more (maximum LOD [MLOD] score 3.4 on 20q11) than males (MLOD score 2.6 on 20p11). With the combined sample, a suggestive LOD score of 2.3 was observed for DSM-IV ND on 20p11. Sex-specific analyses revealed that the signal was driven by females with a maximum LOD score of 3.3 (on 20q11) versus LOD score of 1.3 in males (on 20q13) in the combined sample. No significant linkage signals were obtained for FTND or MaxCigs24.

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Exon–Intron Structure of the Human Neuronal Nicotinic Acetylcholine Receptor α4 Subunit (CHRNA4)

Cited by 48 publications

References 0 publications

Characterization of Human α4β2-Nicotinic Acetylcholine Receptors Stably and Heterologously Expressed in Native Nicotinic Receptor-Null SH-EP1 Human Epithelial Cells

Characterization of Human α4β2-Nicotinic Acetylcholine Receptors Stably and Heterologously Expressed in Native Nicotinic Receptor-Null SH-EP1 Human Epithelial Cells

Characterization of the genomic structure of the human neuronal nicotinic acetylcholine receptor CHRNA5/A3/B4 gene cluster and identification of novel intragenic polymorphisms

Chromosome 20 Shows Linkage With DSM-IV Nicotine Dependence in Finnish Adult Smokers

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