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.
Highlights
Mouse embryonic stem cells, capable of generating most cell types that constitute the entire organism, have made important contributions to our understanding of mammalian biology (Smith, 2001)
In order to apply chemistry to probe the basic biology of neural differentiation from pluripotent stem cells (PSCs), we designed a high content screen to isolate small molecules that can increase the total number of tyrosine hydroxylase (TH)+ neurons derived from Mouse embryonic stem cells (mESCs) monolayer cultures
The system was chosen for several reasons: First, the mESC culture system is an established model for understanding neural development, with much insight gained in recent years (Okano and Temple, 2009; Gaspard and Vanderhaeghen, 2010)
Summary
Mouse embryonic stem cells (mESCs), capable of generating most cell types that constitute the entire organism, have made important contributions to our understanding of mammalian biology (Smith, 2001). Subsequent regional identity and lineageguided differentiation are further regulated by the presence or absence of various morphogens or transcription factors (Lee et al, 2000; Wichterle et al, 2002; Andersson et al, 2006; Martinat et al, 2006). It is unknown whether mechanisms additional to growth factors and transcription regulators direct the differentiation of mESCs into neural lineages. Small organic molecules have proven to be invaluable tools for probing biological mechanisms, owing to their versatile nature and ease of application and removal from the system under study (Stockwell, 2004; Zon and Peterson, 2005). These features make bioactive small molecules highly attractive for therapeutic applications. One critical challenge in small molecule discovery is that the chemical space is infinite, thereby requiring high throughput screening for speed and bioassays that are of sufficient specificity and sensitivity to distinguish active small molecules from background noise
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
