title: Imaging-based chemical screening reveals activity-dependent neural differentiation of pluripotent stem cells.
abstract: Mammalian pluripotent stem cells (PSCs) represent an important venue for understanding basic principles regulating tissue-specific differentiation and discovering new tools that may facilitate clinical applications. Mechanisms that direct neural differentiation of PSCs involve growth factor signaling and transcription regulation. However, it is unknown whether and how electrical activity influences this process. Here we report a high throughput imaging-based screen, which uncovers that selamectin, an anti-helminthic therapeutic compound with reported activity on invertebrate glutamate-gated chloride channels, promotes neural differentiation of PSCs. We show that selamectin's pro-neurogenic activity is mediated by γ2-containing GABAA receptors in subsets of neural rosette progenitors, accompanied by increased proneural and lineage-specific transcription factor expression and cell cycle exit. In vivo, selamectin promotes neurogenesis in developing zebrafish. Our results establish a chemical screening platform that reveals activity-dependent neural differentiation from PSCs. Compounds identified in this and future screening might prove therapeutically beneficial for treating neurodevelopmental or neurodegenerative disorders. DOI:http://dx.doi.org/10.7554/eLife.00508.001.
authors.affiliation: Department of Bioengineering and Therapeutic Science , University of California, San Francisco , San Francisco , United States ; Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research , University of California, San Francisco , San Francisco , United States ; Programs in Human Genetics and Biological Sciences , University of California, San Francisco , San Francisco , United States.
authors.forename: Yaping
authors.initials: Y
authors.lastname: Sun
authors.forename: Zhiqiang
authors.initials: Z
authors.lastname: Dong
authors.forename: Taihao
authors.initials: T
authors.lastname: Jin
authors.forename: Kean-Hooi
authors.initials: KH
authors.lastname: Ang
authors.forename: Miller
authors.initials: M
authors.lastname: Huang
authors.forename: Kelly M
authors.initials: KM
authors.lastname: Haston
authors.forename: Jisong
authors.initials: J
authors.lastname: Peng
authors.forename: Tao P
authors.initials: TP
authors.lastname: Zhong
authors.forename: Steven
authors.initials: S
authors.lastname: Finkbeiner
authors.forename: William A
authors.initials: WA
authors.lastname: Weiss
authors.forename: Michelle R
authors.initials: MR
authors.lastname: Arkin
authors.forename: Lily Y
authors.initials: LY
authors.lastname: Jan
authors.forename: Su
authors.initials: S
authors.lastname: Guo
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D019070:Cell Lineage; D002478:Cells, Cultured; D057166:High-Throughput Screening Assays; D007559:Ivermectin / Q000031:analogs & derivatives / Q000494:pharmacology; D051379:Mice; D009474:Neurons / Q000166:cytology* / Q000187:drug effects; D039904:Pluripotent Stem Cells / Q000166:cytology*; D011963:Receptors, GABA-A / Q000187:drug effects
chemical_list: D011963:Receptors, GABA-A; D007559:Ivermectin; C414354:selamectin
keywords: E. coli; Mouse; Zebrafish; cellular differentiation; chemical genetics; dopaminergic neurons; pluripotent stem cells; small molecule tool
references.citation: Results Probl Cell Differ. 2002;39:27-52
references.pmid: 12357985
references.citation: Trends Neurosci. 2007 Jan;30(1):1-8
references.pmid: 17116335
references.citation: Cell. 2006 Jan 27;124(2):393-405
references.pmid: 16439212
references.citation: J Pharmacol Exp Ther. 1997 Apr;281(1):261-6
references.pmid: 9103505
references.citation: Chem Biol. 2007 Sep;14(9):1019-30
references.pmid: 17884634
references.citation: Nat Methods. 2012 Jun 28;9(7):676-82
references.pmid: 22743772
references.citation: Nat Rev Neurosci. 2012 Jan 18;13(2):94-106
references.pmid: 22251956
references.citation: Drugs. 1991 Oct;42(4):640-58
references.pmid: 1723366
references.citation: Proc Natl Acad Sci U S A. 2006 Feb 21;103(8):2874-9
references.pmid: 16477036
references.citation: Neuron. 1998 Jan;20(1):43-53
references.pmid: 9459441
references.citation: J Neurobiol. 2002 Mar;50(4):305-22
references.pmid: 11891665
references.citation: Nat Biotechnol. 2003 Feb;21(2):183-6
references.pmid: 12524553
references.citation: Proc Natl Acad Sci U S A. 2005 Mar 8;102(10):3840-5
references.pmid: 15738408
references.citation: Nat Biotechnol. 2009 Mar;27(3):275-80
references.pmid: 19252484
references.citation: Arch Insect Biochem Physiol. 2003 Dec;54(4):145-56
references.pmid: 14635176
references.citation: Neuron. 2005 Sep 15;47(6):803-15
references.pmid: 16157276
references.citation: Cell. 2002 Aug 9;110(3):385-97
references.pmid: 12176325
references.citation: Physiol Rev. 2007 Oct;87(4):1215-84
references.pmid: 17928584
references.citation: Nature. 2006 Dec 7;444(7120):707-12
references.pmid: 17151658
references.citation: Proc Natl Acad Sci U S A. 2010 Feb 9;107(6):2687-92
references.pmid: 20133810
references.citation: Science. 1998 Nov 6;282(5391):1145-7
references.pmid: 9804556
references.citation: Nat Rev Neurosci. 2002 Jul;3(7):517-30
references.pmid: 12094208
references.citation: Nat Rev Drug Discov. 2005 Jan;4(1):35-44
references.pmid: 15688071
references.citation: Cell Stem Cell. 2009 Apr 3;4(4):348-58
references.pmid: 19341624
references.citation: J Biol Chem. 2001 Apr 20;276(16):12556-64
references.pmid: 11278873
references.citation: J Physiol. 2003 Aug 1;550(Pt 3):785-800
references.pmid: 12807990
references.citation: J Neurosci. 1982 Jul;2(7):966-71
references.pmid: 6284889
references.citation: Nat Rev Neurosci. 2002 Sep;3(9):715-27
references.pmid: 12209120
references.citation: Biochem J. 1971 May;122(5):759-63
references.pmid: 5129271
references.citation: Neuron. 2010 Jul 29;67(2):321-34
references.pmid: 20670838
references.citation: Nat Rev Neurosci. 2001 Apr;2(4):287-93
references.pmid: 11283751
references.citation: Methods Enzymol. 2012;506:331-60
references.pmid: 22341232
references.citation: Cell Stem Cell. 2008 Jun 5;2(6):602-12
references.pmid: 18522853
references.citation: PLoS One. 2010 Jul 13;5(7):e11528
references.pmid: 20644624
references.citation: Nature. 2004 Oct 14;431(7010):805-10
references.pmid: 15483602
references.citation: Curr Opin Neurobiol. 2009 Apr;19(2):112-9
references.pmid: 19427192
references.citation: Vet Parasitol. 2000 Aug 23;91(3-4):163-76
references.pmid: 10940519
references.citation: J Pharmacol Exp Ther. 2000 Dec;295(3):1051-60
references.pmid: 11082440
references.citation: Nat Neurosci. 2005 Sep;8(9):1179-87
references.pmid: 16116450
references.citation: Annu Rev Cell Dev Biol. 2001;17:435-62
references.pmid: 11687496
references.citation: FASEB J. 2010 Apr;24(4):1218-28
references.pmid: 19959723
references.citation: Stem Cells. 2007 Mar;25(3):562-70
references.pmid: 17110622
references.citation: Am J Stem Cells. 2013 Jun 30;2(2):119-31
references.pmid: 23862100
references.citation: Curr Opin Neurobiol. 2010 Feb;20(1):37-43
references.pmid: 20080043
references.citation: Proc Natl Acad Sci U S A. 2003 Jun 24;100(13):7632-7
references.pmid: 12794184
references.citation: J Mol Evol. 1997 May;44(5):501-8
references.pmid: 9115174
references.citation: Nat Biotechnol. 2000 Jun;18(6):675-9
references.pmid: 10835609
references.citation: Methods Enzymol. 2003;365:327-41
references.pmid: 14696356
references.citation: Nat Rev Neurosci. 2005 Mar;6(3):215-29
references.pmid: 15738957
references.citation: Dev Biol. 2000 Nov 15;227(2):279-93
references.pmid: 11071755
references.citation: J Neurosci Res. 1986;15(2):223-39
references.pmid: 2421007
references.citation: Nature. 2012 Sep 6;489(7414):150-4
references.pmid: 22842902
references.citation: Stem Cells. 2010 Oct;28(10):1741-50
references.pmid: 20734356
references.citation: Nature. 2004 Dec 16;432(7019):846-54
references.pmid: 15602550
references.citation: Genes Dev. 2008 Jan 15;22(2):152-65
references.pmid: 18198334
references.citation: Nat Neurosci. 2010 Aug;13(8):944-50
references.pmid: 20581840
references.citation: Neuron. 1995 Dec;15(6):1287-98
references.pmid: 8845153