High-efficiency channelrhodopsins for fast neuronal stimulation at low light levels.
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High-efficiency channelrhodopsins for fast neuronal stimulation at low light levels. / Berndt, André; Schoenenberger, Philipp; Mattis, Joanna; Tye, Kay M; Deisseroth, Karl; Hegemann, Peter; Oertner, Thomas G.
in: P NATL ACAD SCI USA, Jahrgang 108, Nr. 18, 18, 2011, S. 7595-7600.Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung
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TY - JOUR
T1 - High-efficiency channelrhodopsins for fast neuronal stimulation at low light levels.
AU - Berndt, André
AU - Schoenenberger, Philipp
AU - Mattis, Joanna
AU - Tye, Kay M
AU - Deisseroth, Karl
AU - Hegemann, Peter
AU - Oertner, Thomas G.
PY - 2011
Y1 - 2011
N2 - Channelrhodopsin-2 (ChR2) has become an indispensable tool in neuroscience, allowing precise induction of action potentials with short light pulses. A limiting factor for many optophysiological experiments is the relatively small photocurrent induced by ChR2. We screened a large number of ChR2 point mutants and discovered a dramatic increase in photocurrent amplitude after threonine-to-cysteine substitution at position 159. When we tested the T159C mutant in hippocampal pyramidal neurons, action potentials could be induced at very low light intensities, where currently available channelrhodopsins were unable to drive spiking. Biophysical characterization revealed that the kinetics of most ChR2 variants slows down considerably at depolarized membrane potentials. We show that the recently published E123T substitution abolishes this voltage sensitivity and speeds up channel kinetics. When we combined T159C with E123T, the resulting double mutant delivered fast photocurrents with large amplitudes and increased the precision of single action potential induction over a broad range of frequencies, suggesting it may become the standard for light-controlled activation of neurons.
AB - Channelrhodopsin-2 (ChR2) has become an indispensable tool in neuroscience, allowing precise induction of action potentials with short light pulses. A limiting factor for many optophysiological experiments is the relatively small photocurrent induced by ChR2. We screened a large number of ChR2 point mutants and discovered a dramatic increase in photocurrent amplitude after threonine-to-cysteine substitution at position 159. When we tested the T159C mutant in hippocampal pyramidal neurons, action potentials could be induced at very low light intensities, where currently available channelrhodopsins were unable to drive spiking. Biophysical characterization revealed that the kinetics of most ChR2 variants slows down considerably at depolarized membrane potentials. We show that the recently published E123T substitution abolishes this voltage sensitivity and speeds up channel kinetics. When we combined T159C with E123T, the resulting double mutant delivered fast photocurrents with large amplitudes and increased the precision of single action potential induction over a broad range of frequencies, suggesting it may become the standard for light-controlled activation of neurons.
KW - Animals
KW - Photic Stimulation
KW - Kinetics
KW - Rats
KW - Amino Acid Substitution
KW - Rats, Wistar
KW - Patch-Clamp Techniques
KW - Mutagenesis, Site-Directed
KW - Action Potentials/physiology
KW - Hippocampus/cytology
KW - Light
KW - Neurons/metabolism
KW - Point Mutation/genetics
KW - Pyramidal Cells/metabolism
KW - Rhodopsin/genetics/metabolism
KW - Animals
KW - Photic Stimulation
KW - Kinetics
KW - Rats
KW - Amino Acid Substitution
KW - Rats, Wistar
KW - Patch-Clamp Techniques
KW - Mutagenesis, Site-Directed
KW - Action Potentials/physiology
KW - Hippocampus/cytology
KW - Light
KW - Neurons/metabolism
KW - Point Mutation/genetics
KW - Pyramidal Cells/metabolism
KW - Rhodopsin/genetics/metabolism
M3 - SCORING: Journal article
VL - 108
SP - 7595
EP - 7600
JO - P NATL ACAD SCI USA
JF - P NATL ACAD SCI USA
SN - 0027-8424
IS - 18
M1 - 18
ER -
