Synthetic Light-Activated Ion Channels for Optogenetic Activation and Inhibition
Optogenetic manipulation of cells or living organisms became widely used in neuroscience following the introduction of the light-gated ion channel channelrhodopsin-2 (ChR2). ChR2 is a non-selective cation channel, ideally suited to depolarize and evoke action potentials in neurons. However, its calc...
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doaj-412d2612ff0146a4ba1831dc724d8fb72020-11-25T00:20:59ZengFrontiers Media S.A.Frontiers in Neuroscience1662-453X2018-10-011210.3389/fnins.2018.00643391824Synthetic Light-Activated Ion Channels for Optogenetic Activation and InhibitionSebastian Beck0Jing Yu-Strzelczyk1Dennis Pauls2Oana M. Constantin3Christine E. Gee4Nadine Ehmann5Nadine Ehmann6Nadine Ehmann7Robert J. Kittel8Robert J. Kittel9Robert J. Kittel10Georg Nagel11Shiqiang Gao12Julius-von-Sachs-Institute, University of Würzburg, Würzburg, GermanyJulius-von-Sachs-Institute, University of Würzburg, Würzburg, GermanyNeurobiology and Genetics, Theodor-Boveri Institute, Biocenter, University of Würzburg, Würzburg, GermanyInstitute for Synaptic Physiology, University Medical Center Hamburg-Eppendorf, Hamburg, GermanyInstitute for Synaptic Physiology, University Medical Center Hamburg-Eppendorf, Hamburg, GermanyDepartment of Neurophysiology, Institute of Physiology, University of Würzburg, Würzburg, GermanyDepartment of Animal Physiology, Institute of Biology, Leipzig University, Leipzig, GermanyCarl-Ludwig-Institute for Physiology, Leipzig University, Leipzig, GermanyDepartment of Neurophysiology, Institute of Physiology, University of Würzburg, Würzburg, GermanyDepartment of Animal Physiology, Institute of Biology, Leipzig University, Leipzig, GermanyCarl-Ludwig-Institute for Physiology, Leipzig University, Leipzig, GermanyJulius-von-Sachs-Institute, University of Würzburg, Würzburg, GermanyJulius-von-Sachs-Institute, University of Würzburg, Würzburg, GermanyOptogenetic manipulation of cells or living organisms became widely used in neuroscience following the introduction of the light-gated ion channel channelrhodopsin-2 (ChR2). ChR2 is a non-selective cation channel, ideally suited to depolarize and evoke action potentials in neurons. However, its calcium (Ca2+) permeability and single channel conductance are low and for some applications longer-lasting increases in intracellular Ca2+ might be desirable. Moreover, there is need for an efficient light-gated potassium (K+) channel that can rapidly inhibit spiking in targeted neurons. Considering the importance of Ca2+ and K+ in cell physiology, light-activated Ca2+-permeant and K+-specific channels would be welcome additions to the optogenetic toolbox. Here we describe the engineering of novel light-gated Ca2+-permeant and K+-specific channels by fusing a bacterial photoactivated adenylyl cyclase to cyclic nucleotide-gated channels with high permeability for Ca2+ or for K+, respectively. Optimized fusion constructs showed strong light-gated conductance in Xenopus laevis oocytes and in rat hippocampal neurons. These constructs could also be used to control the motility of Drosophila melanogaster larvae, when expressed in motoneurons. Illumination led to body contraction when motoneurons expressed the light-sensitive Ca2+-permeant channel, and to body extension when expressing the light-sensitive K+ channel, both effectively and reversibly paralyzing the larvae. Further optimization of these constructs will be required for application in adult flies since both constructs led to eclosion failure when expressed in motoneurons.https://www.frontiersin.org/article/10.3389/fnins.2018.00643/fulloptogeneticscalciumpotassiumcAMPbPACCNG channel |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Sebastian Beck Jing Yu-Strzelczyk Dennis Pauls Oana M. Constantin Christine E. Gee Nadine Ehmann Nadine Ehmann Nadine Ehmann Robert J. Kittel Robert J. Kittel Robert J. Kittel Georg Nagel Shiqiang Gao |
spellingShingle |
Sebastian Beck Jing Yu-Strzelczyk Dennis Pauls Oana M. Constantin Christine E. Gee Nadine Ehmann Nadine Ehmann Nadine Ehmann Robert J. Kittel Robert J. Kittel Robert J. Kittel Georg Nagel Shiqiang Gao Synthetic Light-Activated Ion Channels for Optogenetic Activation and Inhibition Frontiers in Neuroscience optogenetics calcium potassium cAMP bPAC CNG channel |
author_facet |
Sebastian Beck Jing Yu-Strzelczyk Dennis Pauls Oana M. Constantin Christine E. Gee Nadine Ehmann Nadine Ehmann Nadine Ehmann Robert J. Kittel Robert J. Kittel Robert J. Kittel Georg Nagel Shiqiang Gao |
author_sort |
Sebastian Beck |
title |
Synthetic Light-Activated Ion Channels for Optogenetic Activation and Inhibition |
title_short |
Synthetic Light-Activated Ion Channels for Optogenetic Activation and Inhibition |
title_full |
Synthetic Light-Activated Ion Channels for Optogenetic Activation and Inhibition |
title_fullStr |
Synthetic Light-Activated Ion Channels for Optogenetic Activation and Inhibition |
title_full_unstemmed |
Synthetic Light-Activated Ion Channels for Optogenetic Activation and Inhibition |
title_sort |
synthetic light-activated ion channels for optogenetic activation and inhibition |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Neuroscience |
issn |
1662-453X |
publishDate |
2018-10-01 |
description |
Optogenetic manipulation of cells or living organisms became widely used in neuroscience following the introduction of the light-gated ion channel channelrhodopsin-2 (ChR2). ChR2 is a non-selective cation channel, ideally suited to depolarize and evoke action potentials in neurons. However, its calcium (Ca2+) permeability and single channel conductance are low and for some applications longer-lasting increases in intracellular Ca2+ might be desirable. Moreover, there is need for an efficient light-gated potassium (K+) channel that can rapidly inhibit spiking in targeted neurons. Considering the importance of Ca2+ and K+ in cell physiology, light-activated Ca2+-permeant and K+-specific channels would be welcome additions to the optogenetic toolbox. Here we describe the engineering of novel light-gated Ca2+-permeant and K+-specific channels by fusing a bacterial photoactivated adenylyl cyclase to cyclic nucleotide-gated channels with high permeability for Ca2+ or for K+, respectively. Optimized fusion constructs showed strong light-gated conductance in Xenopus laevis oocytes and in rat hippocampal neurons. These constructs could also be used to control the motility of Drosophila melanogaster larvae, when expressed in motoneurons. Illumination led to body contraction when motoneurons expressed the light-sensitive Ca2+-permeant channel, and to body extension when expressing the light-sensitive K+ channel, both effectively and reversibly paralyzing the larvae. Further optimization of these constructs will be required for application in adult flies since both constructs led to eclosion failure when expressed in motoneurons. |
topic |
optogenetics calcium potassium cAMP bPAC CNG channel |
url |
https://www.frontiersin.org/article/10.3389/fnins.2018.00643/full |
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