title: miRNAome analysis of the mammalian neuronal nicotinic acetylcholine receptor gene family.
abstract: Nicotine binds to and activates a family of ligand-gated ion channels, neuronal nicotinic acetylcholine receptors (nAChRs). Chronic nicotine exposure alters the expression of various nAChR subtypes, which likely contributes to nicotine dependence; however, the underlying mechanisms regulating these changes remain unclear. A growing body of evidence indicates that microRNAs (miRNAs) may be involved in nAChR regulation. Using bioinformatics, miRNA library screening, site-directed mutagenesis, and gene expression analysis, we have identified a limited number of miRNAs that functionally interact with the 3'-untranslated regions (3' UTRs) of mammalian neuronal nAChR subunit genes. In silico analyses revealed specific, evolutionarily conserved sites within the 3' UTRs through which the miRNAs regulate gene expression. Mutating these sites disrupted miRNA regulation confirming the in silico predictions. In addition, the miRNAs that target nAChR 3' UTRs are expressed in mouse brain and are regulated by chronic nicotine exposure. Furthermore, we show that expression of one of these miRNAs, miR-542-3p, is modulated by nicotine within the mesocorticolimbic reward pathway. Importantly, overexpression of miR-542-3p led to a decrease in the protein levels of its target, the nAChR β2 subunit. Bioinformatic analysis suggests that a number of the miRNAs play a general role in regulating cholinergic signaling. Our results provide evidence for a novel mode of nicotine-mediated regulation of the mammalian nAChR gene family.
authors.affiliation: Brudnick Neuropsychiatric Research Institute, Department of Psychiatry, University of Massachusetts Medical School, Worcester, Massachusetts 01604, USA Brudnick Neuropsychiatric Research Institute, Department of Psychiatry, University of Massachusetts Medical School, Worcester, Massachusetts 01604, USA.
authors.forename: Eric M
authors.initials: EM
authors.lastname: Hogan
authors.affiliation: Brudnick Neuropsychiatric Research Institute, Department of Psychiatry, University of Massachusetts Medical School, Worcester, Massachusetts 01604, USA UMMS MD/PhD Program, University of Massachusetts Medical School, Worcester, Massachusetts 01604, USA.
authors.forename: Alison P
authors.initials: AP
authors.lastname: Casserly
authors.affiliation: Brudnick Neuropsychiatric Research Institute, Department of Psychiatry, University of Massachusetts Medical School, Worcester, Massachusetts 01604, USA Brudnick Neuropsychiatric Research Institute, Department of Psychiatry, University of Massachusetts Medical School, Worcester, Massachusetts 01604, USA.
authors.forename: Michael D
authors.initials: MD
authors.lastname: Scofield
authors.affiliation: Brudnick Neuropsychiatric Research Institute, Department of Psychiatry, University of Massachusetts Medical School, Worcester, Massachusetts 01604, USA.
authors.forename: Zhongming
authors.initials: Z
authors.lastname: Mou
authors.affiliation: Brudnick Neuropsychiatric Research Institute, Department of Psychiatry, University of Massachusetts Medical School, Worcester, Massachusetts 01604, USA.
authors.forename: Rubing
authors.initials: R
authors.lastname: Zhao-Shea
authors.affiliation: Brudnick Neuropsychiatric Research Institute, Department of Psychiatry, University of Massachusetts Medical School, Worcester, Massachusetts 01604, USA.
authors.forename: Chris W
authors.initials: CW
authors.lastname: Johnson
authors.affiliation: Brudnick Neuropsychiatric Research Institute, Department of Psychiatry, University of Massachusetts Medical School, Worcester, Massachusetts 01604, USA Brudnick Neuropsychiatric Research Institute, Department of Psychiatry, University of Massachusetts Medical School, Worcester, Massachusetts 01604, USA UMMS MD/PhD Program, University of Massachusetts Medical School, Worcester, Massachusetts 01604, USA.
authors.forename: Andrew R
authors.initials: AR
authors.lastname: Tapper
authors.affiliation: Brudnick Neuropsychiatric Research Institute, Department of Psychiatry, University of Massachusetts Medical School, Worcester, Massachusetts 01604, USA Brudnick Neuropsychiatric Research Institute, Department of Psychiatry, University of Massachusetts Medical School, Worcester, Massachusetts 01604, USA UMMS MD/PhD Program, University of Massachusetts Medical School, Worcester, Massachusetts 01604, USA paul.gardner@umassmed.edu.
authors.forename: Paul D
authors.initials: PD
authors.lastname: Gardner
mesh_terms: D020413:3' Untranslated Regions; D000818:Animals; D001921:Brain / Q000378:metabolism; D005786:Gene Expression Regulation / Q000235:genetics; D007473:Ion Channels / Q000096:biosynthesis* / Q000235:genetics; D008024:Ligands; D008322:Mammals; D051379:Mice; D035683:MicroRNAs / Q000096:biosynthesis* / Q000235:genetics; D016297:Mutagenesis, Site-Directed; D009538:Nicotine / Q000378:metabolism*; D009928:Organ Specificity; D011978:Receptors, Nicotinic / Q000096:biosynthesis / Q000235:genetics*; D015398:Signal Transduction / Q000235:genetics
chemical_list: D020413:3' Untranslated Regions; D007473:Ion Channels; D008024:Ligands; D035683:MicroRNAs; D011978:Receptors, Nicotinic; D009538:Nicotine
keywords: gene expression; miR-542-3p; miRNA analysis; nicotinic acetylcholine receptor