The distribution of mRNA encoded by a new member of the neuronal nicotinic acetylcholine receptor gene family (α5) in the rat central nervous system

doi:10.1016/0006-8993(90)90248-A

Brain Research

Volume 526, Issue 1, 27 August 1990, Pages 45-53

https://doi.org/10.1016/0006-8993(90)90248-A Get rights and content

Abstract

The cellular localization of transcripts for a new putative agonist-binding subunit of the neuronal nicotinic acetylcholine receptor (nAChR), α5, was examined using in situ hybridization in the rat central nervous system (CNS). α5 subunit mRNA was localized to a small number of regions when compared with two of the other known agonist-binding subunits, α3 and α4. α5 mRNA is expressed at relatively high levels in neurons of the subiculum (pyramidal layer), presubiculum and parasubiculum (layers IV and VI), which are components of the hippocampal formation, in the substantia nigra pars compacta and ventral tegmental area, in the interpeduncular nucleus, and in the dorsal motor nucleus of the vagus nerve. Moderate hybridization signals were detected in neurons of the isocortex (layer VIb), anterior olfactory nucleus, trigeminal ganglion, superior olivary complex, nucleus of the solitary tract, and area postrema. No hybridization above background levels was seen in the amygdala, septum, thalamus, hypothalamus, or cerebellum. These results suggest that the α5 subunit differs from other known agonist-binding subunits in its distribution.

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Chrna5 and lynx prototoxins identify acetylcholine super-responder subplate neurons
2023, iScience
Attention depends on cholinergic excitation of prefrontal neurons but is sensitive to perturbation of α5-containing nicotinic receptors encoded by Chrna5. However, Chrna5-expressing (Chrna5+) neurons remain enigmatic, despite their potential as a target to improve attention. Here, we generate complex transgenic mice to probe Chrna5+ neurons and their sensitivity to endogenous acetylcholine. Through opto-physiological experiments, we discover that Chrna5+ neurons contain a distinct population of acetylcholine super-responders. Leveraging single-cell transcriptomics, we discover molecular markers conferring subplate identity on this subset. We determine that Chrna5+ super-responders express a unique complement of GPI-anchored lynx prototoxin genes (Lypd1, Ly6g6e, and Lypd6b), predicting distinct nicotinic receptor regulation. To manipulate lynx regulation of endogenous nicotinic responses, we developed a pharmacological strategy guided by transcriptomic predictions. Overall, we reveal Chrna5-Cre mice as a transgenic tool to target the diversity of subplate neurons in adulthood, yielding new molecular strategies to manipulate their cholinergic activation relevant to attention disorders.
Mutation of the α5 nicotinic acetylcholine receptor subunit increases ethanol and nicotine consumption in adolescence and impacts adult drug consumption
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Alcohol and nicotine are commonly used during adolescence, establishing long-lasting neuroplastic alterations that influence subsequent drug use and abuse. Drinking- and smoking-related traits have been extensively associated with variation in CHRNA5 – the gene that encodes the α5 subunit of neuronal nicotinic acetylcholine receptors (nAChRs). The single nucleotide polymorphism (SNP) rs16969968 in CHRNA5 encodes an amino acid substitution (D398N) that alters the function and pharmacokinetics of α5-containing nAChR. When expressed in rodents, this variant results in increased ethanol and nicotine operant self-administration. How disruption of α5-containing nAChRs influences adolescent ethanol and nicotine intake, and how it modulates interactions between these drugs has not been previously explored. In the present study, we examined volitional ethanol and nicotine consumption in adolescent mice (post-natal day 30–43) of both sexes with mutated (SNP) or lacking (KO) the α5 nAChR subunit. The effect of adolescent alcohol or nicotine exposure on home cage consumption of the opposite drug in adulthood and its modulation by Chrna5 mutation and sex were examined. During adolescence, we found that α5 nAChR disruption increases nicotine intake in mice of both sexes, but the effect on alcohol intake was only observed in females. The sex-specific increase in alcohol consumption in α5 SNP and KO was replicated in adulthood. The effect of adolescent alcohol or nicotine exposure on subsequent intake of the opposite drug in adulthood is modulated by sex and Chrna5 mutation. These observations suggest sex differences in the genetic architecture of alcohol dependence, and modulators of alcohol and nicotine interactions.
Linking the CHRNA5 SNP to drug abuse liability: From circuitry to cellular mechanisms
2021, Neuropharmacology
Genetics are known to be a significant risk factor for drug abuse. In human populations, the single nucleotide polymorphism (SNP) D398N in the gene CHRNA5 has been associated with addiction to nicotine, opioids, cocaine, and alcohol. In this paper, we review findings from studies in humans, rodent models, and cell lines and provide evidence that collectively suggests that the Chrna5 SNP broadly influences the response to drugs of abuse in a manner that is not substance-specific. This finding has important implications for our understanding of the role of the cholinergic system in reward and addiction vulnerability.
This article is part of the special issue on ‘Vulnerabilities to Substance Abuse.’
Alterations in nicotinic receptor alpha5 subunit gene differentially impact early and later stages of cocaine addiction: a translational study in transgenic rats and patients
2021, Progress in Neurobiology
Citation Excerpt :
The VTA, dorsal prefrontal cortex and NAcc shell are crucial to cocaine-induced reinstatement while the central nucleus of the amygdala and bed nucleus of the stria terminalis are necessary for stress-induced reinstatement (Kalivas and McFarland, 2003). In this line, α5 is expressed at relatively high levels in the VTA and NAcc, but absent in the amygdala (Wada et al., 1990). The lack of effect of Chrna5 deletion on cue- and stress-induced relapse seems in accordance with the physiological absence of α5 containing nAChRs in brain areas specifically involved in these modalities of relapse.
Cocaine addiction is a chronic and relapsing disorder with an important genetic component. Human candidate gene association studies showed that the single nucleotide polymorphism (SNP) rs16969968 in the α5 subunit (α5SNP) of nicotinic acetylcholine receptors (nAChRs), previously associated with increased tobacco dependence, was linked to a lower prevalence of cocaine use disorder (CUD). Three additional SNPs in the α5 subunit, previously shown to modify α5 mRNA levels, were also associated with CUD, suggesting an important role of the subunit in this pathology. To investigate the link between this subunit and CUD, we submitted rats knockout for the α5 subunit gene (α5KO), or carrying the α5SNP, to cocaine self-administration (SA) and showed that the acquisition of cocaine-SA was impaired in α5SNP rats while α5KO rats exhibited enhanced cocaine-induced relapse associated with altered neuronal activity in the nucleus accumbens. In addition, we observed in a human cohort of patients with CUD that the α5SNP was associated with a slower transition from first cocaine use to CUD. We also identified a novel SNP in the β4 nAChR subunit, part of the same gene cluster in the human genome and potentially altering CHRNA5 expression, associated with shorter time to relapse to cocaine use in patients.
In conclusion, the α5SNP is protective against CUD by influencing early stages of cocaine exposure while CHRNA5 expression levels may represent a biomarker for the risk to relapse to cocaine use. Drugs modulating α5 containing nAChR activity may thus represent a novel therapeutic strategy against CUD.
Genetic susceptibility to nicotine addiction: Advances and shortcomings in our understanding of the CHRNA5/A3/B4 gene cluster contribution
2020, Neuropharmacology
Citation Excerpt :
Taken together, these preclinical results fit well with the human situation where the α5SNP is associated with an increased risk for NA, lower ratings of aversive effect, and delayed smoking cessation (Bierut et al., 2008; Chen et al., 2015; Jensen et al., 2015; Saccone et al., 2007). The α5 nAChR subunit is expressed in discrete regions of the rodent central nervous system (CNS), including the striatum where it regulates DA transmission (Exley et al., 2012), the cerebral cortex, cerebellum, thalamus, hippocampus, substantia nigra (SN), VTA and mHb, with the highest concentration found in the IPN (Forget et al., 2018; Hsu et al., 2013; Marks et al., 1992; Sheffield et al., 2000; Wada et al., 1990). According to the databases listed in part 2.1.1, rodent Chrna5 expression partly follows human CHRNA5 expression, with an overall higher expression in the hindbrain (notably the cerebellum) than in the forebrain, but with constantly detectable (although lower) expression in the cortex and in the thalamus.
Over the last decade, robust human genetic findings have been instrumental in elucidating the heritable basis of nicotine addiction (NA). They highlight coding and synonymous polymorphisms in a cluster on chromosome 15, encompassing the CHRNA5, CHRNA3 and CHRNB4 genes, coding for three subunits of the nicotinic acetylcholine receptor (nAChR). They have inspired an important number of preclinical studies, and will hopefully lead to the definition of novel drug targets for treating NA. Here, we review these candidate gene and genome-wide association studies (GWAS) and their direct implication in human brain function and NA-related phenotypes. We continue with a description of preclinical work in transgenic rodents that has led to a mechanistic understanding of several of the genetic hits. We also highlight important issues with regards to CHRNA3 and CHRNB4 where we are still lacking a dissection of their role in NA, including even in preclinical models. We further emphasize the use of human induced pluripotent stem cell-derived models for the analysis of synonymous and intronic variants on a human genomic background. Finally, we indicate potential avenues to further our understanding of the role of this human genetic variation.
This article is part of the special issue on ‘Contemporary Advances in Nicotine Neuropharmacology’.
Cell-Genotype Specific Effects of Mecp2 Mutation on Spontaneous and Nicotinic Acetylcholine Receptor-Evoked Currents in Medial Prefrontal Cortical Pyramidal Neurons in Female Rett Model Mice
2019, Neuroscience
Rett syndrome (RTT) is a neurodevelopmental disorder caused by mutation in the X-linked MECP2 gene. Random X-inactivation produces a mosaic of mutant (MT) and wild-type (WT) neurons in female Mecp2 +/− (het) mice. Many RTT symptoms are alleviated by increasing activity in medial prefrontal cortex (mPFC) in RTT model mice (Howell et al., 2017). Using a GFP-MeCP2 fusion protein to distinguish WT from MT pyramidal neurons in mPFC we found cell autonomous (cell genotype specific) and non-autonomous effects of MeCP2 deficiency on spontaneous excitatory/inhibitory balance, nicotinic acetylcholine receptor (nAChR) currents and evoked activity. MT Layer 5 and 6 (L5, L6) neurons of male nulls, and MT L6 of het mice had reduced spontaneous excitatory synaptic input compared to WT in wild-type male (WTm), female (WTf) and het mice. Inhibitory synaptic charge in MT L6 equaled WT in 2–4-month hets. At 6–7 months inhibitory charge in WT in het slices was increased compared to both MT in het and WT in WTf; however, in hets the excitatory/inhibitory charge ratio was still greater in WT compared to MT. nAChR currents were reduced in L6 of nulls and MT L6 in het slices compared to WT neurons of het, WTm and WTf. At 2–4 months, ACh perfusion increased frequency of inhibitory currents to L6 neurons equally in all genotypes but increased excitatory inputs to MT and WT in hets less than WT in WTfs. Unexpectedly ACh perfusion evoked greater sustained IPSC and EPSC input to L5 neurons of nulls compared to WTm.

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^*: Present address: Laboratory of Neurochemistry, NINDS, NIH, Bldg. 36, Room 4D20, Bethesda, MD 20892, U.S.A.

^**: Present address: Department of Pharmacology, Washington University School of Medicine, St. Louis, MO 63110, U.S.A.

^***: Present address: Baylor College of Medicine, Division of Nueroscience, One Baylor Plaza, Houston, TX 77030, U.S.A.

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The distribution of mRNA encoded by a new member of the neuronal nicotinic acetylcholine receptor gene family (α5) in the rat central nervous system

Abstract

Dev. Biol.

Neuron

J. Biol. Chem.

Cell

Brain Research

J. Biol. Chem.

Brain Research

Brain Res. Bull.

The nucleus paragigantocellularis lateralis in the rat

Anat. Embryol.

The Brain Stem of the Cat. A Cytoarchitectonic Atlas with Stereotaxic Coordinates

Viscerotropic representation of the upper alimentary tract in the medulla oblongata in the rat: the nucleus ambiguus

J. Comp. Neurol.

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

Nature

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

Nicotinic binding in rat brain: autoradiographic comparison of [3H]acetylcholine, [3H]nicotine, and [125I]-α-bungarotoxin

J. Neurosci.

The strychnine-binding subunit of the glycine receptor shows homology with nicotinic acetylcholine receptors

Nature

Cytoarchitecture, fiber connections, and some histochemical aspects of the interpeduncular nucleus in the rat

J. Comp. Neurol.

The distribution of mRNA encoded by a new member of the neuronal nicotinic acetylcholine receptor gene family (α₅) in the rat central nervous system

Nicotinic binding in rat brain: autoradiographic comparison of [³H]acetylcholine, [³H]nicotine, and [¹²⁵I]-α-bungarotoxin