Abstract
Optogenetics is a growing technique which allows manipulation of biological events simply by illumination. The technique is appreciated especially in the neuroscience field because of its availability in controlling neuronal functions. A light-gated cation channel, Cr_ChR2 from Chlamydomonas reinhardtii, is the first and mostly applied to optogenetics for activating neuronal excitability. In addition, the molecular mechanism of Cr_ChR2 has been intensively studied by electrophysiology, spectroscopy, X-ray structural studies, etc. Novel cation channelrhodopsins from Guillardia theta, namely, Gt_CCR1–4, were discovered in 2016 and 2017. These channelrhodopsins are more homologous to haloarchaeal rhodopsins, particularly the proton pumps. Thus these cryptophyte-type light-gated cation channels are structurally and mechanistically distinct from chlorophyte channelrhodopsin such as Cr_ChR2. We here compared the photocurrent properties, cation selectivity, and kinetics between well-known Cr_ChR2 and Gt_CCR4. The light sensitivity of Gt_CCR4 is significantly higher than that of Cr_ChR2, while the channel open lifetime is in the same range as that of Cr_ChR2. Gt_CCR4 shows high Na+ selectivity in which the selectivity ratio for Na+ was 37-fold larger than that for Cr_ChR2, which primarily conducts H+. On the other hand, Gt_CCR4 conducted almost no H+ and no Ca2+ under physiological conditions. Other unique features and the applicability of Gt_CCR4 for optogenetics were discussed.
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Funding
This work was supported by the Japanese Ministry of Education, Culture, Sports, Science and Technology (25104009, 15H02391 to H.K and 18 K06109 to S.P.T.), a JST CREST grant (JPMJCR1753 to H.K.), and a JST PRESTO grant (JPMJPR1688 to S.P.T). S.H. is a Research Fellow of the Japan Society for the Promotion of Science (JSPS Research Fellow).
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Hososhima, S., Shigemura, S., Kandori, H. et al. Novel optogenetics tool: Gt_CCR4, a light-gated cation channel with high reactivity to weak light. Biophys Rev 12, 453–459 (2020). https://doi.org/10.1007/s12551-020-00676-7
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DOI: https://doi.org/10.1007/s12551-020-00676-7