title: Functional Characterization of Acetylcholine Receptors Expressed in Human Neurons Differentiated from Hippocampal Neural Stem/Progenitor Cells.
abstract: Neurotransmission mediated by acetylcholine receptors (AChRs) plays an important role in learning and memory functions in the hippocampus. Impairment of the cholinergic system contributes to Alzheimer's disease (AD), indicating the importance of AChRs as drug targets for AD. To improve the success rates for AD drug development, human cell models that mimic the target brain region are important. Therefore, we characterized the functional expression of nicotinic and muscarinic AChRs (nAChRs and mAChRs, respectively) in human hippocampal neurons differentiated from hippocampal neural stem/progenitor cells (HIP-009 cells). Intracellular calcium flux in 4-week differentiated HIP-009 cells demonstrated that the cells responded to acetylcholine, nicotine, and muscarine in a concentration-dependent manner (EC50= 13.4 ± 0.5, 6.0 ± 0.4, and 35.0 ± 2.5 µM, respectively). In addition, assays using subtype-selective compounds revealed that major AD therapeutic target AChR subtypes-α7 and α4β2 nAChRs, as well as M1and M3mAChRs-were expressed in the cells. Furthermore, neuronal network analysis demonstrated that potentiation of M3mAChRs inhibits the spontaneous firing of HIP-009 neurons. These results indicate that HIP-009 cells are physiologically relevant for AD drug screening and hence are loadstars for the establishment of in vitro AD models.
authors: 1 Next Generation Systems Core Function Unit, Eisai Product Creation Systems, Eisai Co., Ltd., Tsukuba, Ibaraki, Japan.|2 Department of Genomics-Based Drug Discovery, Doctoral Program in Clinical Sciences, Graduate School of Comprehensive Human Science, University of Tsukuba, Tsukuba, Ibaraki, Japan.
authors: Kazuyuki
authors: K
authors: Fukushima
authors: 1 Next Generation Systems Core Function Unit, Eisai Product Creation Systems, Eisai Co., Ltd., Tsukuba, Ibaraki, Japan.
authors: Kazuto
authors: K
authors: Yamazaki
authors: 2 Department of Genomics-Based Drug Discovery, Doctoral Program in Clinical Sciences, Graduate School of Comprehensive Human Science, University of Tsukuba, Tsukuba, Ibaraki, Japan.|3 Global Cardiovascular Assessment, Biopharmaceutical Assessment Core Function Unit, Eisai Product Creation Systems, Eisai Co., Ltd., Tsukuba, Ibaraki, Japan.
authors: Norimasa
authors: N
authors: Miyamoto
authors: 2 Department of Genomics-Based Drug Discovery, Doctoral Program in Clinical Sciences, Graduate School of Comprehensive Human Science, University of Tsukuba, Tsukuba, Ibaraki, Japan.|3 Global Cardiovascular Assessment, Biopharmaceutical Assessment Core Function Unit, Eisai Product Creation Systems, Eisai Co., Ltd., Tsukuba, Ibaraki, Japan.
authors: Kohei
authors: K
authors: Sawada
mesh_terms: D000109:Acetylcholine / Q000378:metabolism; D000544:Alzheimer Disease / Q000188:drug therapy* / Q000235:genetics; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics*; D016503:Drug Delivery Systems / Q000379:methods*; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D006624:Hippocampus / Q000166:cytology / Q000187:drug effects; D006801:Humans; D009116:Muscarine / Q000378:metabolism; D058953:Neural Stem Cells / Q000166:cytology / Q000378:metabolism; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D009538:Nicotine / Q000378:metabolism; D018408:Patch-Clamp Techniques; D043643:Receptor, Muscarinic M1 / Q000037:antagonists & inhibitors / Q000235:genetics; D043587:Receptor, Muscarinic M3 / Q000037:antagonists & inhibitors / Q000235:genetics; D011950:Receptors, Cholinergic / Q000187:drug effects / Q000235:genetics; D011978:Receptors, Nicotinic / Q000187:drug effects / Q000235:genetics; D013234:Stem Cells / Q000166:cytology / Q000378:metabolism; D009435:Synaptic Transmission / Q000187:drug effects* / Q000235:genetics
chemical_list: D043643:Receptor, Muscarinic M1; D043587:Receptor, Muscarinic M3; D011950:Receptors, Cholinergic; D011978:Receptors, Nicotinic; D009538:Nicotine; D009116:Muscarine; D000109:Acetylcholine
keywords: human hippocampal neurons; multielectrode array; muscarinic acetylcholine receptors; neuronal network; nicotinic acetylcholine receptors