The functional diversity of the neuronal nicotinic acetylcholine receptors is increased by a novel subunit: β4

doi:10.1016/0896-6273(89)90207-9

Neuron

Volume 3, Issue 4, October 1989, Pages 487-496

https://doi.org/10.1016/0896-6273(89)90207-9 Get rights and content

Abstract

A new nicotinic acetylcholine receptor (nAChR) subunit, β4 was identified by screening a rat genomic library. In situ hybridization histochemistry revealed expression of the β4 gene in the medial habenula of adult rat brains. The primary structure of this subunit was deduced from a cDNA clone isolated from a PC12 cDNA library. Functional nAChRs were detected in Xenopus oocytes injected in pairwise combinations with in vitro synthesized RNAs encoding β4 and either the α2, 0, or α4 subunit. Unlike the a3β2 receptor, the α3β4 receptor is not blocked by bungarotoxin 3.1, indicating that the β subunit can affect the sensitivity of neuronal nAChRs to this toxin. These results extend the functional diversity of nicotinic receptors in the nervous system.

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Citation Excerpt :
Here, we show that the density of [3H]epibatidine binding to high nicotine-affinity CHRNs and levels of mRNA for CHRNB2 are not altered in BA 24 from subjects with MDD or BD. The absence of detectable CHRNB4 in BA 24 is consistent with the lower cortical expression of CHRNB4 compared to CHRNB2 reported in rats (Duvoisin et al., 1989; Hill et al., 1993) supporting our use of CHRNB2 as a surrogate measure of high-nicotine affinity CHRN. These data argue that the changes in CHRM2 in the ACC from patients with mood disorders are not associated with changes in levels of CHRNs caused by changed prevailing levels of acetylcholine.
We have previously reported reduced expression of the cholinergic autoreceptor CHRM2 in Brodmann's Area (BA) 24 of the anterior cingulate cortex from subjects with major depressive disorder (MDD) and bipolar disorder (BD), consistent with a hypercholinergic state. This led us to investigate whether levels of the high affinity nicotinic acetylcholine receptors are also altered in BA 24.
We measured the binding levels of a high-affinity nicotinic receptor-selective radioligand, [³H]epibatidine, in BA 24 from subjects with MDD (n = 20), BD (n = 18) and age- and sex-matched controls (n = 20). We used qPCR to measure mRNA expression of the high affinity nicotinic acetylcholine receptor subunit CHRNB2 in these subjects.
[³H]Epibatidine binding density and CHRNB2 mRNA expression were not significantly altered in either MDD or BD compared to control levels. While validating reference genes for our qPCR experiments, we found that the mRNA levels of 3 putative reference genes, TFB1M, PPIA and SNCA, were increased in MDD but not BD compared to controls. Further investigations in other cortical regions showed that these changes were specific to BA24.
Cohort size and available patient data were limited due to standard constraints associated with post-mortem studies.
Our data suggest that decreased CHRM2 in BA24 in mood disorders is not associated with a corresponding change in high affinity nicotinic acetylcholine receptor expression. Our findings of increased TFB1M, PPIA and SNCA expression in MDD point to a broader derangement of several homeostatic pathways in MDD that are distinct from BD.
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Citation Excerpt :
The first clones to be isolated from neuronal tissues included three such alpha subunits (α2, α3, and α4) as well as three subunits lacking the vicinal cysteines, two of which would form functional nAChRs when co-expressed with the muscle genes α1, γ, and ε, leading these subunits to be designated as neuronal beta subunits (β2, β3, and β4), although β3 did not co-assemble with the muscle subunits, and its function remained a mystery for a number of years, as was also the case for the α5 and α6 subunits, which were cloned later (Heinemann et al., 1990). It was subsequently shown that the α2, α3, or α4 subunits could form functional nAChRs in the Xenopus oocyte heterologous expression system if co-expressed with β2 (Boulter et al., 1987; Deneris et al., 1988; Wada et al., 1988) or β4 subunits (Duvoisin et al., 1989). In situ hybridization showed that the α2, α3, and α4 subunits had distinct, largely non-overlapping patterns of expression in the rat brain, with α4 expressed more widely than the other two subunits, and also that β2 was widely expressed in a pattern that overlapped the expression of all three α subunits (Wada et al., 1989).
Postsynaptic nAChRs in the peripheral nervous system are critical for neuromuscular and autonomic neurotransmission. Pre- and peri-synaptic nAChRs in the brain modulate neurotransmission and are responsible for the addictive effects of nicotine. Subtypes of nAChRs in lymphocytes and non-synaptic locations may modulate inflammation and other cellular functions. All AChRs that function as ligand-gated ion channels are formed from five homologous subunits organized to form a central cation channel whose opening is regulated by ACh bound at extracellular subunit interfaces. nAChR subtype subunit composition can range from α7 homomers to α4β2α6β2β3 heteromers. Subtypes differ in affinities for ACh and other agonists like nicotine and in efficiencies with which their channels are opened and desensitized. Subtypes also differ in affinities for antagonists and for positive and negative allosteric modulators. Some agonists are “silent” with respect to channel opening, and AChRs may be able to signal metabotropic pathways by releasing G-proteins independent of channel opening. Electrophysiological studies that can resolve single-channel openings and molecular genetic approaches have allowed characterization of the structures of ligand binding sites, the cation channel, and the linkages between them, as well as the organization of AChR subunits and their contributions to function. Crystallography and cryo-electron-microscopy are providing increasing insights into the structures and functions of AChRs. However, much remains to be learned about both AChR structure and function, the in vivo functional roles of some AChR subtypes, and the development of better pharmacological tools directed at AChRs to treat addiction, pain, inflammation, and other medically important issues.
This article is part of the special issue on ‘Contemporary Advances in Nicotine Neuropharmacology’.
The role of acetylcholine and its receptors in retinal processing
2016, The Curated Reference Collection in Neuroscience and Biobehavioral Psychology
Acetylcholine (ACh) is synthesized in the retina by two populations of amacrine cells. The effects of ACh are mediated by muscarinic (mAChR) and nicotinic acetylcholine receptors (nAChRs). The three β and nine α nAChR subunits and 5 mAChR subtypes identified in the vertebrate retina display distinctive physiological and pharmacological characteristics. Amacrine and ganglion cells in the retina express one or more AChR subtypes and there is evidence for expression by subpopulations of bipolar cells as well. ACh can affect the response properties of ganglion cells through receptors expressed by the ganglion cells themselves as well receptors on upstream cells.
Differential expression of the beta4 neuronal nicotinic receptor subunit affects tolerance development and nicotinic binding sites following chronic nicotine treatment
2015, Pharmacology Biochemistry and Behavior
The role of neuronal nicotinic acetylcholine receptors (nAChR) containing the β4 subunit in tolerance development and nicotinic binding site levels following chronic nicotine treatment was investigated. Mice differing in expression of the β4-nAChR subunit [wild-type (β4⁺⁺), heterozygote (β4⁺⁻) and null mutant (β4⁻⁻)] were chronically treated for 10 days with nicotine (0, 0.5, 1.0, 2.0 or 4.0 mg/kg/h) by constant intravenous infusion. Chronic nicotine treatment elicited dose-dependent tolerance development. β4⁻⁻ mice developed significantly more tolerance than either β4⁺⁺ or β4⁺⁻ mice which was most evident following treatment with 4.0 mg/kg/h nicotine. Subsets of [¹²⁵I]-epibatidine binding were measured in several brain regions. Deletion of the β4 subunit had little effect on initial levels of cytisine-sensitive [¹²⁵I]-epibatidine binding (primarily α4β2-nAChR sites) or their response (generally increased binding) to chronic nicotine treatment. In contrast, β4 gene-dose-dependent decreases in expression 5IA-85380 resistant [¹²⁵I]-epibatidine binding sites (primarily β4*-nAChR) were observed. While these β4*-nAChR sites were generally resistant to regulation by chronic nicotine treatment, significant increases in binding were noted for habenula and hindbrain. Comparison of previously published tolerance development in β2⁻⁻ mice (less tolerance) to that of β4⁻⁻ mice (more tolerance) supports a differential role for these receptor subtypes in regulating tolerance following chronic nicotine treatment.
The Role of Nicotine in Schizophrenia
2015, International Review of Neurobiology
Citation Excerpt :
Partial loss of α3 results in decreased susceptibility to nicotine-induced seizures, but normal hypolocomotor response to nicotine (Salas, Cook, Bassetto, & De Biasi, 2004). Expression of β4 is limited to the medial habenula, olfactory bulbs, and a handful other areas (Duvoisin, Deneris, Patrick, & Heinemann, 1989; Salas, Pieri, Fung, Dani, & De Biasi, 2003). While studies of the CHRNA5–CHRNA3–CHRNB4 region have mostly focused on CHRNA5 or CHRNA3, associations have been shown between polymorphisms in CHRNB4 and some aspects of smoking (Schlaepfer et al., 2008).
Schizophrenia is associated with by severe disruptions in thought, cognition, emotion, and behavior. Patients show a marked increase in rates of smoking and nicotine dependence relative to nonaffected individuals, a finding commonly ascribed to the potential ameliorative effects of nicotine on symptom severity and cognitive impairment. Indeed, many studies have demonstrated improvement in patients following the administration of nicotine. Such findings have led to an increased emphasis on the development of therapeutic agents to target the nicotinic system as well as increasing the impetus to understand the genetic basis for nicotinic dysfunction in schizophrenia. The goal of this review article is to provide a critical summary of evidence for the role of the nicotinic system in schizophrenia. The first part will review the role of nicotine in normalization of primary dysfunctions and endophenotypical changes found in schizophrenia. The second part will provide a summary of genetic evidence linking polymorphisms in nicotinic receptor genes to smoking and schizophrenia. The third part will summarize attempts to treat schizophrenia using agents specifically targeting nicotinic and nicotinic receptor subtypes. Although currently available antipsychotic treatments are generally able to manage some aspects of schizophrenia (e.g., positive symptoms) they fail to address several other critically effected aspects of the disease. As such, the search for novel mechanisms to treat this disease is necessary.
Impact of human D398N single nucleotide polymorphism on intracellular calcium response mediated by α3β4α5 nicotinic acetylcholine receptors
2012, Neuropharmacology
The human CHRNA5 D398N polymorphism (rs16969968) causes an aspartic acid to asparagine change in the nicotinic acetylcholine receptor (nAChR) α5 subunit gene. The N398 variant of CHRNA5 is linked to increased risk for nicotine dependence. In this study, we explored the effect of the CHRNA5 D398N polymorphism on the properties of human α3β4* nicotinic acetylcholine receptors in human embryonic kidney (HEK) cells.
Addition of either D398 or N398 variant of α5 subunit in the α3β4* receptor did not affect total [¹²⁵I]-epibatidine binding or surface expression of the receptor. However, addition of α5^D398 into α3β4* receptor decreased the maximal response to agonist without significantly affecting EC₅₀ in aequorin intracellular calcium assay. α3β4α5^N398 nAChRs showed further decreased maximal response. The differences in agonist efficacy between the receptor subtypes were found to be dependent upon the concentration of external calcium but independent of external sodium. Moreover, activation of α3β4α5 nAChRs led to significantly greater intracellular calcium release from IP₃ stores relative to α3β4 nAChRs although no effect of the α5 polymorphism was observed. Finally, inclusion of the α5 variant caused a small shift to the left in IC₅₀ for some of the antagonists tested, depending upon α5 variant but did not affect sensitivity of α3β4* receptors to desensitization in response to incubation with nicotine.
In conclusion, addition of either variant of α5 into an α3β4α5 receptor similarly effects receptor pharmacology and function. However, the N398 variant exhibits a reduced response to agonists when extracellular calcium is high and it may lead to distinct downstream cellular signaling.

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^∗: Present address: Division of Neurosciences, Baylor College of Medicine, Houston, Texas, 77030.

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Neuron

ArticleThe functional diversity of the neuronal nicotinic acetylcholine receptors is increased by a novel subunit: β4

Abstract

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Isolation of a cDNA clone coding for a possible neural nicotinic acetylcholine receptor alpha-subunit

Nature

Functional expression of two neural nicotinic acetylcholine receptors from cDNA clones identifies a gene family

A method for isolation of intact, translationally active ribonucleic acid

DNA

The acetylcholine receptor: an “allosteric” membrane protein

Harvey Lect.

Kappa-bungarotoxin: a probe for the neuronal nicotinic receptor in the avian ciliary ganglion

Brain Res.

Translation of eukaryotic messenger RNA in Xenopus oocytes

A comprehensive set of sequence analysis programs for the VAX

Nucl. Acids Res.

Mapping of brain areas expressing RNA homologous to two different acetylcholine receptor a-subunit cDNAs

Affinity labeling of neuronal acetylcholine receptor subunits with an a-neurotoxin that blocks receptor function

J. Neurosci.

Article
The functional diversity of the neuronal nicotinic acetylcholine receptors is increased by a novel subunit: β4