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Optical inhibition of zebrafish behavior with anion channelrhodopsins

Gadisti Aisha Mohamed, Ruey-Kuang Cheng, View ORCID ProfileJoses Ho, View ORCID ProfileSeetha Krishnan, View ORCID ProfileFarhan Mohammad, View ORCID ProfileAdam Claridge-Chang, View ORCID ProfileSuresh Jesuthasan
doi: https://doi.org/10.1101/158899
Gadisti Aisha Mohamed
1Lee Kong Chian School of Medicine, Nanyang Technological University, Singapore.
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Ruey-Kuang Cheng
1Lee Kong Chian School of Medicine, Nanyang Technological University, Singapore.
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Joses Ho
2Institute of Molecular and Cell Biology, Singapore.
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Seetha Krishnan
3Graduate School for Integrative Sciences and Engineering, National University of Singapore.
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Farhan Mohammad
4Duke-NUS Medical School, Singapore
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Adam Claridge-Chang
2Institute of Molecular and Cell Biology, Singapore.
4Duke-NUS Medical School, Singapore
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Suresh Jesuthasan
1Lee Kong Chian School of Medicine, Nanyang Technological University, Singapore.
2Institute of Molecular and Cell Biology, Singapore.
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Abstract

In behavioral analysis, optical electrical silencing provides a way to test neuronal necessity. Two light-gated anion channels, GtACR1 and GtACR2, have recently been shown—in neuronal culture and in Drosophila—to inhibit neurons potently. Here, we test the usefulness of these channels in zebrafish. When the GtACRs were expressed in motor neurons and actuated with blue or green light, fish spontaneous movement was inhibited. In GtACR1-expressing fish, only 3 µW/mm2 of light was sufficient to have an effect; GtACR2, which is poorly trafficked, required stronger illumination. After light offset, GtACR-expressing fish movement increased; this suggested that termination of light-induced neural inhibition may lead to depolarization. Consistent with this, two-photon imaging of spinal neurons showed that intracellular calcium also increased following light offset. The activity elicited at light offset needs to be taken into consideration in experimental design, although this property may help provide insight into the effects of stimulating a circuit transiently. These results show that GtACR1 and GtACR2 can be used to optically inhibit neurons in zebrafish, and thus to test neural circuit necessity.

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Posted July 04, 2017.
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Optical inhibition of zebrafish behavior with anion channelrhodopsins
Gadisti Aisha Mohamed, Ruey-Kuang Cheng, Joses Ho, Seetha Krishnan, Farhan Mohammad, Adam Claridge-Chang, Suresh Jesuthasan
bioRxiv 158899; doi: https://doi.org/10.1101/158899
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Optical inhibition of zebrafish behavior with anion channelrhodopsins
Gadisti Aisha Mohamed, Ruey-Kuang Cheng, Joses Ho, Seetha Krishnan, Farhan Mohammad, Adam Claridge-Chang, Suresh Jesuthasan
bioRxiv 158899; doi: https://doi.org/10.1101/158899

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