Biophysical characterization of light-gated ion channels using planar automated patch clamp
Channelrhodopsins (ChRs) are proteins that guide phototaxis in protists and exhibit light-gated channel conductance when their genes are heterologously expressed in mammalian cells. ChRs are widely used as molecular tools to control neurons and cardiomyocytes with light (optogenetics). Cation- and a...
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2022-08-01
|
| Series: | Frontiers in Molecular Neuroscience |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fnmol.2022.976910/full |
| _version_ | 1818504443940831232 |
|---|---|
| author | Elena G. Govorunova Oleg A. Sineshchekov Leonid S. Brown John L. Spudich |
| author_facet | Elena G. Govorunova Oleg A. Sineshchekov Leonid S. Brown John L. Spudich |
| author_sort | Elena G. Govorunova |
| collection | DOAJ |
| description | Channelrhodopsins (ChRs) are proteins that guide phototaxis in protists and exhibit light-gated channel conductance when their genes are heterologously expressed in mammalian cells. ChRs are widely used as molecular tools to control neurons and cardiomyocytes with light (optogenetics). Cation- and anion-selective ChRs (CCRs and ACRs, respectively) enable stimulation and inhibition of neuronal activity by depolarization and hyperpolarization of the membrane, respectively. More than 400 natural ChR variants have been identified so far, and high-throughput polynucleotide sequencing projects add many more each year. However, electrophysiological characterization of new ChRs lags behind because it is mostly done by time-consuming manual patch clamp (MPC). Here we report using a high-throughput automated patch clamp (APC) platform, SyncroPatch 384i from Nanion Technologies, for ChR research. We find that this instrument can be used for determination of the light intensity dependence and current-voltage relationships in ChRs and discuss its advantages and limitations. |
| first_indexed | 2024-12-10T21:37:11Z |
| format | Article |
| id | doaj.art-b4eccaa5540c48518a48586fa578c475 |
| institution | Directory Open Access Journal |
| issn | 1662-5099 |
| language | English |
| last_indexed | 2024-12-10T21:37:11Z |
| publishDate | 2022-08-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Molecular Neuroscience |
| spelling | doaj.art-b4eccaa5540c48518a48586fa578c4752022-12-22T01:32:37ZengFrontiers Media S.A.Frontiers in Molecular Neuroscience1662-50992022-08-011510.3389/fnmol.2022.976910976910Biophysical characterization of light-gated ion channels using planar automated patch clampElena G. Govorunova0Oleg A. Sineshchekov1Leonid S. Brown2John L. Spudich3Department of Biochemistry and Molecular Biology, Center for Membrane Biology, The University of Texas Health Science Center at Houston McGovern Medical School, Houston, TX, United StatesDepartment of Biochemistry and Molecular Biology, Center for Membrane Biology, The University of Texas Health Science Center at Houston McGovern Medical School, Houston, TX, United StatesBiophysics Interdepartmental Group, Department of Physics, University of Guelph, Guelph, ON, CanadaDepartment of Biochemistry and Molecular Biology, Center for Membrane Biology, The University of Texas Health Science Center at Houston McGovern Medical School, Houston, TX, United StatesChannelrhodopsins (ChRs) are proteins that guide phototaxis in protists and exhibit light-gated channel conductance when their genes are heterologously expressed in mammalian cells. ChRs are widely used as molecular tools to control neurons and cardiomyocytes with light (optogenetics). Cation- and anion-selective ChRs (CCRs and ACRs, respectively) enable stimulation and inhibition of neuronal activity by depolarization and hyperpolarization of the membrane, respectively. More than 400 natural ChR variants have been identified so far, and high-throughput polynucleotide sequencing projects add many more each year. However, electrophysiological characterization of new ChRs lags behind because it is mostly done by time-consuming manual patch clamp (MPC). Here we report using a high-throughput automated patch clamp (APC) platform, SyncroPatch 384i from Nanion Technologies, for ChR research. We find that this instrument can be used for determination of the light intensity dependence and current-voltage relationships in ChRs and discuss its advantages and limitations.https://www.frontiersin.org/articles/10.3389/fnmol.2022.976910/fullchannelrhodopsinsoptogeneticspotassium channelskalium channelrhodopsinsautomated patch clamplight-gated channels |
| spellingShingle | Elena G. Govorunova Oleg A. Sineshchekov Leonid S. Brown John L. Spudich Biophysical characterization of light-gated ion channels using planar automated patch clamp Frontiers in Molecular Neuroscience channelrhodopsins optogenetics potassium channels kalium channelrhodopsins automated patch clamp light-gated channels |
| title | Biophysical characterization of light-gated ion channels using planar automated patch clamp |
| title_full | Biophysical characterization of light-gated ion channels using planar automated patch clamp |
| title_fullStr | Biophysical characterization of light-gated ion channels using planar automated patch clamp |
| title_full_unstemmed | Biophysical characterization of light-gated ion channels using planar automated patch clamp |
| title_short | Biophysical characterization of light-gated ion channels using planar automated patch clamp |
| title_sort | biophysical characterization of light gated ion channels using planar automated patch clamp |
| topic | channelrhodopsins optogenetics potassium channels kalium channelrhodopsins automated patch clamp light-gated channels |
| url | https://www.frontiersin.org/articles/10.3389/fnmol.2022.976910/full |
| work_keys_str_mv | AT elenaggovorunova biophysicalcharacterizationoflightgatedionchannelsusingplanarautomatedpatchclamp AT olegasineshchekov biophysicalcharacterizationoflightgatedionchannelsusingplanarautomatedpatchclamp AT leonidsbrown biophysicalcharacterizationoflightgatedionchannelsusingplanarautomatedpatchclamp AT johnlspudich biophysicalcharacterizationoflightgatedionchannelsusingplanarautomatedpatchclamp |