Optical inhibition of larval zebrafish behaviour with anion channelrhodopsins

BMC Biology
Gadisti Aisha MohamedSuresh Jesuthasan

Abstract

Optical silencing of activity provides a way to test the necessity of neurons in behaviour. Two light-gated anion channels, GtACR1 and GtACR2, have recently been shown to potently inhibit activity in cultured mammalian neurons and in Drosophila. Here, we test the usefulness of these channels in larval zebrafish, using spontaneous coiling behaviour as the assay. When the GtACRs were expressed in spinal neurons of embryonic zebrafish and actuated with blue or green light, 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 slightly stronger illumination. No inhibition was seen in non-expressing siblings. After light offset, the movement of GtACR-expressing fish increased, which suggested that termination of light-induced neural inhibition may lead to activation. Consistent with this, two-photon imaging of spinal neurons showed that blue light inhibited spontaneous activity in spinal neurons of GtACR1-expressing fish, and that the level of intracellular calcium increased following light offset. These results show that GtACR1 and GtACR2 can be used to optically inhibit neurons in larval zebrafish with high efficiency. Th...Continue Reading

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Nov 7, 2018·Nature Communications·Yinth Andrea Bernal SierraDietmar Schmitz
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Dec 29, 2020·Neuron·António M FernandesHerwig Baier
Mar 8, 2018·Chemical Reviews·Hideki KandoriSatoshi P Tsunoda

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Methods Mentioned

BETA
transgenic

Key Resources (RRID) Mentioned

SCR_002285

Software Mentioned

Fiji
MicroManager
iChloC

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