title: Pharmacological Characterisation of Nicotinic Acetylcholine Receptors Expressed in Human iPSC-Derived Neurons.
abstract: Neurons derived from human induced pluripotent stem cells (iPSCs) represent a potentially valuable tool for the characterisation of neuronal receptors and ion channels. Previous studies on iPSC-derived neuronal cells have reported the functional characterisation of a variety of receptors and ion channels, including glutamate receptors, γ-aminobutyric acid (GABA) receptors and several voltage-gated ion channels. In the present study we have examined the expression and functional properties of nicotinic acetylcholine receptors (nAChRs) in human iPSC-derived neurons. Gene expression analysis indicated the presence of transcripts encoding several nAChR subunits, with highest levels detected for α3-α7, β1, β2 and β4 subunits (encoded by CHRNA3-CHRNA7, CHRNB1, CHRNB2 and CHRNB4 genes). In addition, similarly high transcript levels were detected for the truncated dupα7 subunit transcript, encoded by the partially duplicated gene CHRFAM7A, which has been associated with psychiatric disorders such as schizophrenia. The functional properties of these nAChRs have been examined by calcium fluorescence and by patch-clamp recordings. The data obtained suggest that the majority of functional nAChRs expressed in these cells have pharmacological properties typical of α7 receptors. Large responses were induced by a selective α7 agonist (compound B), in the presence of the α7-selective positive allosteric modulator (PAM) PNU-120596, which were blocked by the α7-selective antagonist methyllycaconitine (MLA). In addition, a small proportion of the neurons express nAChRs with properties typical of heteromeric (non-α7 containing) nAChR subtypes. These cells therefore represent a great tool to advance our understanding of the properties of native human nAChRs, α7 in particular.
authors.affiliation: Department of Neuroscience, Physiology & Pharmacology, University College London, London, United Kingdom.
authors.forename: Anna
authors.initials: A
authors.lastname: Chatzidaki
authors.affiliation: Lilly Research Centre, Eli Lilly and Company, Windlesham, Surrey, United Kingdom.
authors.forename: Antoine
authors.initials: A
authors.lastname: Fouillet
authors.affiliation: Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, Indiana 46285, United States of America.
authors.forename: Jingling
authors.initials: J
authors.lastname: Li
authors.affiliation: Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, Indiana 46285, United States of America.
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Dage
authors.affiliation: Department of Neuroscience, Physiology & Pharmacology, University College London, London, United Kingdom.
authors.forename: Neil S
authors.initials: NS
authors.lastname: Millar
authors.affiliation: Lilly Research Centre, Eli Lilly and Company, Windlesham, Surrey, United Kingdom.
authors.forename: Emanuele
authors.initials: E
authors.lastname: Sher
authors.affiliation: Lilly Research Centre, Eli Lilly and Company, Windlesham, Surrey, United Kingdom.
authors.forename: Daniel
authors.initials: D
authors.lastname: Ursu
mesh_terms: D000157:Aconitine / Q000031:analogs & derivatives / Q000494:pharmacology; D002478:Cells, Cultured; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000187:drug effects / Q000378:metabolism*; D007555:Isoxazoles / Q000494:pharmacology; D009474:Neurons / Q000166:cytology* / Q000378:metabolism; D010671:Phenylurea Compounds / Q000494:pharmacology; D011978:Receptors, Nicotinic / Q000235:genetics*; D059010:Single-Cell Analysis
chemical_list: C508388:1-(5-chloro-2,4-dimethoxyphenyl)-3-(5-methylisoxazol-3-yl)urea; D007555:Isoxazoles; D010671:Phenylurea Compounds; D011978:Receptors, Nicotinic; C054634:methyllycaconitine; D000157:Aconitine