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Driving Neurogenesis in Neural Stem Cells with High Sensitivity Optogenetics
Daniel Boon Loong Teh1,2, Ankshita Prasad3, Wenxuan Jiang4
1Department of Biochemistry, National University of Singapore, 28 Medical Drive, 4-44, Singapore, 117456, Singapore. danielteh@nus.edu.sg.
Neuromolecular Medicine
|October 10, 2019
Summary
Optogenetic stimulation of neural stem cells (NSCs) was enhanced using a novel light-sensitive protein and a simple system. This method promotes neuron differentiation and reveals transcriptome changes, offering a safer, more efficient tool for neuroscience research.
Area of Science:
- Neuroscience
- Stem Cell Biology
- Optogenetics
Background:
- Optogenetics allows neural stem cell (NSC) activity modulation for studying neurogenesis and regulating differentiation.
- Current methods using channelrhodopsin-2 (ChR2) require high light intensity and complex systems.
- Transcriptome changes from optogenetic NSC stimulation remain largely uncharacterized.
Purpose of the Study:
- To develop a more sensitive and efficient optogenetic system for NSC manipulation.
- To investigate the impact of optogenetic stimulation on NSC differentiation and gene expression.
- To establish a low-irradiance optical stimulation (OS) system for transformed NSCs (SFO-NSCs).
Main Methods:
- Engineered SFO-NSCs using a non-viral piggyBac transposon system to express a light-sensitive step-function opsin (SFO) variant (ChRFR(C167A)).
- Developed a simple, low-irradiance OS-incubation system.
- Analyzed c-fos mRNA expression as an indicator of activity-dependent response.
- Performed transcriptome analysis to identify gene expression changes.
Main Results:
- Daily OS induced c-fos mRNA expression in differentiating SFO-NSCs, confirming activity-dependent responses.
- Optogenetically stimulated SFO-NSCs exhibited enhanced neuron-like differentiation with more elongated axons compared to controls.
- Transcriptome analysis revealed significant gene expression changes related to axonal remodeling, synaptic plasticity, and microenvironment modulation, including upregulation of Ca2+-related genes.
Conclusions:
- The developed SFO-NSC system offers enhanced light sensitivity and requires lower irradiance for effective stimulation.
- This approach facilitates activity-dependent differentiation and provides insights into transcriptome-level alterations.
- The non-viral, low-irradiance method presents a promising tool for stem cell transplantation studies with reduced carcinogenicity and improved tissue penetration.
