Kinetic evaluation of photosensitivity in bi-stable variants of chimeric channelrhodopsins

PloS One
Shoko HososhimaHiromu Yawo

Abstract

Channelrhodopsin-1 and 2 (ChR1 and ChR2) form cation channels that are gated by light through an unknown mechanism. We tested the DC-gate hypothesis that C167 and D195 are involved in the stabilization of the cation-permeable state of ChRWR/C1C2 which consists of TM1-5 of ChR1 and TM6-7 of ChR2 and ChRFR which consists of TM1-2 of ChR1 and TM3-7 of ChR2. The cation permeable state of each ChRWR and ChRFR was markedly prolonged in the order of several tens of seconds when either C167 or D195 position was mutated to alanine (A). Therefore, the DC-gate function was conserved among these chimeric ChRs. We next investigated the kinetic properties of the ON/OFF response of these bi-stable ChR mutants as they are important in designing the photostimulation protocols for the optogenetic manipulation of neuronal activities. The turning-on rate constant of each photocurrent followed a linear relationship to 0-0.12 mW mm(-2) of blue LED light or to 0-0.33 mW mm(-2) of cyan LED light. Each photocurrent of bi-stable ChR was shut off to the non-conducting state by yellow or orange LED light in a manner dependent on the irradiance. As the magnitude of the photocurrent was mostly determined by the turning-on rate constant and the irradiation t...Continue Reading

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Citations

Nov 11, 2015·Scientific Reports·Shoko HososhimaHiromu Yawo
Nov 20, 2016·PloS One·Mohammad Razuanul HoqueHiromu Yawo
Oct 9, 2019·Neuromolecular Medicine·Daniel Boon Loong TehAngelo All
Sep 4, 2018·Frontiers in Neuroscience·Toshifumi AsanoHiromu Yawo
May 16, 2018·The Journal of Physiological Sciences : JPS·Kyo KoizumiHiromu Yawo
Apr 5, 2018·Scientific Reports·Hiroyuki IgarashiHiromu Yawo
Sep 28, 2018·Nature Communications·Kazumasa OdaOsamu Nureki

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Methods Mentioned

BETA
transfection
targeted mutation
irradiate

Software Mentioned

pCLAMP10

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