ArticleJournal of visualized experiments : JoVE2017
Whole-cell Patch-clamp Recordings for Electrophysiological Determination of Ion Selectivity in Channelrhodopsins.
Article in Journal of visualized experiments : JoVE, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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Who cites it
13 citing papers in PubMed.
- Multicolor photoreactions of the red light-activated channelrhodopsin Chrimson.Proceedings of the National Academy of Sciences of the United States of America · 2025Article
- Robust optogenetic inhibition with red-light-sensitive anion-conducting channelrhodopsins.eLife · 2024Article
- A mechanism of lysosomal calcium entry.Science advances · 2024Article
- 3D printing-based frugal manufacturing of glass pipettes for minimally invasive delivery of therapeutics to the brain.Neuroprotection (Chichester, England) · 2023Article
- WiChR, a highly potassium-selective channelrhodopsin for low-light one- and two-photon inhibition of excitable cells.Science advances · 2022Article
- Aion is a bistable anion-conducting channelrhodopsin that provides temporally extended and reversible neuronal silencing.Communications biology · 2022Article
- BiPOLES is an optogenetic tool developed for bidirectional dual-color control of neurons.Nature communications · 2021Article
- Porous Graphene Oxide Decorated Ion Selective Electrode for Observing Across-Cytomembrane Ion Transport.Sensors (Basel, Switzerland) · 2020Article
- MerMAIDs: a family of metagenomically discovered marine anion-conducting and intensely desensitizing channelrhodopsins.Nature communications · 2019Article
- Unifying photocycle model for light adaptation and temporal evolution of cation conductance in channelrhodopsin-2.Proceedings of the National Academy of Sciences of the United States of America · 2019Article
- Crystal structure of the red light-activated channelrhodopsin Chrimson.Nature communications · 2018Article
- Electrical properties, substrate specificity and optogenetic potential of the engineered light-driven sodium pump eKR2.Scientific reports · 2018Article
- Anion-conducting channelrhodopsins with tuned spectra and modified kinetics engineered for optogenetic manipulation of behavior.Scientific reports · 2017Article
Corrections and comments
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Authors and funding
4 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Over the past decade, channelrhodopsins became indispensable in neuroscientific research where they are used as tools to non-invasively manipulate electrical processes in target cells. In this context, ion selectivity of a channelrhodopsin is of particular importance. This article describes the investigation of chloride selectivity for a recently identified anion-conducting channelrhodopsin of Proteomonas sulcata via electrophysiological patch-clamp recordings on HEK293 cells. The experimental procedure for measuring light-gated photocurrents demands a fast switchable - ideally monochromatic - light source coupled into the microscope of an otherwise conventional patch-clamp setup. Preparative procedures prior to the experiment are outlined involving preparation of buffered solutions, considerations on liquid junction potentials, seeding and transfection of cells, and pulling of patch pipettes. The actual recording of current-voltage relations to determine the reversal potentials for different chloride concentrations takes place 24 h to 48 h after transfection. Finally, electrophysiological data are analyzed with respect to theoretical considerations of chloride conduction.
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