An improved chloride-conducting channelrhodopsin for light-induced inhibition of neuronal activity in vivo
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An improved chloride-conducting channelrhodopsin for light-induced inhibition of neuronal activity in vivo. / Wietek, Jonas; Beltramo, Riccardo; Scanziani, Massimo; Hegemann, Peter; Oertner, Thomas G; Simon Wiegert, J.
In: SCI REP-UK, Vol. 5, 07.10.2015, p. 14807.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
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T1 - An improved chloride-conducting channelrhodopsin for light-induced inhibition of neuronal activity in vivo
AU - Wietek, Jonas
AU - Beltramo, Riccardo
AU - Scanziani, Massimo
AU - Hegemann, Peter
AU - Oertner, Thomas G
AU - Simon Wiegert, J
PY - 2015/10/7
Y1 - 2015/10/7
N2 - Channelrhodopsins are light-gated cation channels that have been widely used for optogenetic stimulation of electrically excitable cells. Replacement of a glutamic acid in the central gate with a positively charged amino acid residue reverses the ion selectivity and produces chloride-conducting ChRs (ChloCs). Expressed in neurons, published ChloCs produced a strong shunting effect but also a small, yet significant depolarization from the resting potential. Depending on the state of the neuron, the net result of illumination might therefore be inhibitory or excitatory with respect to action potential generation. Here we report two additional amino acid substitutions that significantly shift the reversal potential of improved ChloC (iChloC) to the reversal potential of endogenous GABAA receptors. As a result, light-evoked membrane depolarization was strongly reduced and spike initiation after current injection or synaptic stimulation was reliably inhibited in iChloC-transfected neurons in vitro. In the primary visual cortex of anesthetized mice, activation of iChloC suppressed spiking activity evoked by visual stimulation. Due to its high operational light sensitivity, iChloC makes it possible to inhibit neurons in a large volume of brain tissue from a small, point-like light source.
AB - Channelrhodopsins are light-gated cation channels that have been widely used for optogenetic stimulation of electrically excitable cells. Replacement of a glutamic acid in the central gate with a positively charged amino acid residue reverses the ion selectivity and produces chloride-conducting ChRs (ChloCs). Expressed in neurons, published ChloCs produced a strong shunting effect but also a small, yet significant depolarization from the resting potential. Depending on the state of the neuron, the net result of illumination might therefore be inhibitory or excitatory with respect to action potential generation. Here we report two additional amino acid substitutions that significantly shift the reversal potential of improved ChloC (iChloC) to the reversal potential of endogenous GABAA receptors. As a result, light-evoked membrane depolarization was strongly reduced and spike initiation after current injection or synaptic stimulation was reliably inhibited in iChloC-transfected neurons in vitro. In the primary visual cortex of anesthetized mice, activation of iChloC suppressed spiking activity evoked by visual stimulation. Due to its high operational light sensitivity, iChloC makes it possible to inhibit neurons in a large volume of brain tissue from a small, point-like light source.
U2 - 10.1038/srep14807
DO - 10.1038/srep14807
M3 - SCORING: Journal article
C2 - 26443033
VL - 5
SP - 14807
JO - SCI REP-UK
JF - SCI REP-UK
SN - 2045-2322
ER -