TY - JOUR
T1 - Driving Neurogenesis in Neural Stem Cells with High Sensitivity Optogenetics
AU - Teh, Daniel Boon Loong
AU - Prasad, Ankshita
AU - Jiang, Wenxuan
AU - Zhang, Nianchen
AU - Wu, Yang
AU - Yang, Hyunsoo
AU - Han, Sanyang
AU - Yi, Zhigao
AU - Yeo, Yanzhuang
AU - Ishizuka, Toru
AU - Wong, Limsoon
AU - Thakor, Nitish
AU - Yawo, Hiromu
AU - Liu, Xiaogang
AU - All, Angelo
N1 - This work is supported by R-175-000-137-112 (Singapore Ministry of Education Tier 1), R-175-000-136-305 (Singapore Agency for Science, Technology and Research A*STAR - Japan Science and Technology Agency JST), Grants-in-Aid for Scientific Research (KAKENHI) from the Ministry of Education, Culture, Sports, Science and Technology (MEXT), Japan (25670103 to HY), and JST, Strategic International Collaborative Research Program, SICORP (15657509A to HY).
PY - 2020/3
Y1 - 2020/3
N2 - Optogenetic stimulation of neural stem cells (NSCs) enables their activity-dependent photo-modulation. This provides a spatio-temporal tool for studying activity-dependent neurogenesis and for regulating the differentiation of the transplanted NSCs. Currently, this is mainly driven by viral transfection of channelrhodopsin-2 (ChR2) gene, which requires high irradiance and complex in vivo/vitro stimulation systems. Additionally, despite the extensive application of optogenetics in neuroscience, the transcriptome-level changes induced by optogenetic stimulation of NSCs have not been elucidated yet. Here, we made transformed NSCs (SFO-NSCs) stably expressing one of the step-function opsin (SFO)-variants of chimeric channelrhodopsins, ChRFR(C167A), which is more sensitive to blue light than native ChR2, via a non-viral transfection system using piggyBac transposon. We set up a simple low-irradiance optical stimulation (OS)-incubation system that induced c-fos mRNA expression, which is activity-dependent, in differentiating SFO-NSCs. More neuron-like SFO-NCSs, which had more elongated axons, were differentiated with daily OS than control cells without OS. This was accompanied by positive/negative changes in the transcriptome involved in axonal remodeling, synaptic plasticity, and microenvironment modulation with the up-regulation of several genes involved in the Ca2+-related functions. Our approach could be applied for stem cell transplantation studies in tissue with two strengths: lower carcinogenicity and less irradiance needed for tissue penetration.
AB - Optogenetic stimulation of neural stem cells (NSCs) enables their activity-dependent photo-modulation. This provides a spatio-temporal tool for studying activity-dependent neurogenesis and for regulating the differentiation of the transplanted NSCs. Currently, this is mainly driven by viral transfection of channelrhodopsin-2 (ChR2) gene, which requires high irradiance and complex in vivo/vitro stimulation systems. Additionally, despite the extensive application of optogenetics in neuroscience, the transcriptome-level changes induced by optogenetic stimulation of NSCs have not been elucidated yet. Here, we made transformed NSCs (SFO-NSCs) stably expressing one of the step-function opsin (SFO)-variants of chimeric channelrhodopsins, ChRFR(C167A), which is more sensitive to blue light than native ChR2, via a non-viral transfection system using piggyBac transposon. We set up a simple low-irradiance optical stimulation (OS)-incubation system that induced c-fos mRNA expression, which is activity-dependent, in differentiating SFO-NSCs. More neuron-like SFO-NCSs, which had more elongated axons, were differentiated with daily OS than control cells without OS. This was accompanied by positive/negative changes in the transcriptome involved in axonal remodeling, synaptic plasticity, and microenvironment modulation with the up-regulation of several genes involved in the Ca2+-related functions. Our approach could be applied for stem cell transplantation studies in tissue with two strengths: lower carcinogenicity and less irradiance needed for tissue penetration.
KW - Calcium Signaling
KW - Cell Line, Transformed
KW - Channelrhodopsins/biosynthesis
KW - DNA Transposable Elements
KW - Gene Expression Regulation/radiation effects
KW - Gene Ontology
KW - Genes, Reporter
KW - Genes, fos
KW - Nerve Tissue Proteins/biosynthesis
KW - Neural Stem Cells/cytology
KW - Neurogenesis/radiation effects
KW - Neuronal Plasticity/radiation effects
KW - Optogenetics
KW - RNA, Messenger/biosynthesis
KW - Transcriptome/radiation effects
KW - Up-Regulation/radiation effects
U2 - 10.1007/s12017-019-08573-3
DO - 10.1007/s12017-019-08573-3
M3 - Journal article
C2 - 31595404
SN - 1535-1084
VL - 22
SP - 139
EP - 149
JO - NeuroMolecular Medicine
JF - NeuroMolecular Medicine
IS - 1
ER -