Robust optogenetic inhibition with red-light-sensitive anion-conducting channelrhodopsins
- Institut für Biologie, Experimentelle Biophysik, Humboldt-Universität zu Berlin, Germany
- Faculty of Biology, Technion – Israel Institute of Technology, Israel
- Research Group Neuromodulatory Networks, Leibniz Institute for Neurobiology, Germany
- Aligning Science Across Parkinson's (ASAP) Collaborative Research Network, United States
- Center for Behavioral Brain Sciences, CBBS, Germany
Figures
Figure 1
Phylogeny and electrophysiological characterization of identified anion-conducting channelrhodopsins (ACRs).
(A) Phylogenetic relationships among known channelrhodopsins (ChRs) subfamilies with the described ChRs written in black and blue font. The color of the clades indicates ion selectivity: red for …
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Figure 1—source data 1
Source data for electrophysiological analysis of identified anion-conducting channelrhodopsins presented in Figure 1B, C, E, F.
- https://cdn.elifesciences.org/articles/90100/elife-90100-fig1-data1-v1.xlsx
Figure 2
Color-tuning of MsACR1.
(A) Structural model of the MsACR1 retinal binding pocket. Residues close to the β-ionone ring are labeled in cyan, and those close to the retinal Schiff base (RSB) are labeled purple. Residue S218, …
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Figure 2—source data 1
Source data for electrophysiological analysis of color-tuning in MsACR1 presented in Figure 2C-F.
- https://cdn.elifesciences.org/articles/90100/elife-90100-fig2-data1-v1.xlsx
Figure 3
Silencing of neuronal activity in dissociated hippocampal neurons using MsACR1 and raACR.
(A) Confocal image of a hippocampal neuron 10 days after viral transduction expressing MsACR1-mCerulean under control of the hSynapsin1 (hSyn) promoter. The magnified portion of the image of a …
Figure 4
Inhibition of locomotion in rodents in vivo using MsACR1 and raACR.
(A) Confocal images of coronal brain slices confirm the expression of raACR in the primary motor cortex (M1) and the accurate placement of the optical cannula. A close-up of the injection site …
Figure 5
Activation of st-MsACR1 and st-raACR in PV-Cre neurons in M1 leads to complex changes in firing rate.
(A) Illustration of the injection strategy, viral constructs, optrode design, and implantation in PV-Cre mice. The diagram shows the targeting of M1 with hSyn-DIO-st-MsACR1-mCerulean, …
Appendix 1—figure 1
Maximum-likelihood phylogenetic reconstruction of relationships among known channelrhodopsins.
The sequences chosen for the analysis were taken from the Catalog of Natural Channelrhodopsins (Rozenberg, 2023). Names based on the taxonomic affiliation of the organisms possessing the …
Appendix 1—figure 2
Alignment of the rhodopsin domains of the channelrhodopsins (ChRs) tested in the current study and chosen reference proteins.
The alignment was constructed using structural information using the crystal structures of BR (PDB: 1AP9), ChR2 (6EID), Chrimson (5ZIH), and GtACR1 (6CSM) and AlphaFold structures for the ChRs …
Appendix 1—figure 3
Light sensitivity and ion selectivity of identified ACRs.
(A) Full action spectra of identified ACRs. Shown are mean ± standard deviation connected by lines. (B) Light titration of identified ACRs at light intensities between 38 nW/mm2 and 3.2 mW/mm2 using …
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Appendix 1—figure 3—source data 1
Source data for electrophysiological analysis to determine light sensitivity and ion selectivity of identified ACRs presented in Appendix 1—figure 3A-C, E, F.
- https://cdn.elifesciences.org/articles/90100/elife-90100-app1-fig3-data1-v1.xlsx
Appendix 1—figure 4
Example photocurrent traces of MsACR1 wild-type and mutants.
Photocurrent traces of MsACR1 and all mutants tested in this study, measured in ND7/23 cells. Photocurrents were elicited with 560 nm light (gray bars) at membrane voltages between −80 and +40 mV in …
Appendix 1—figure 5
The red-light activity of ACRs.
(A) Structural model of the MsACR1. Residue side chains and the all-trans-retinal chromophore are shown as sticks except for G123 and G186, where the Cɑ is shown as a sphere. Oxygen is colored in …
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Appendix 1—figure 5—source data 1
Source data for electrophysiological analysis to determine red-light activity of ACRs presented in Appendix 1—figure 5B and C.
- https://cdn.elifesciences.org/articles/90100/elife-90100-app1-fig5-data1-v1.xlsx
Appendix 1—figure 6
Example traces of MsACR1 and raACR in neurons.
Voltage traces of MsACR1, raACR, and their soma-targeted versions expressing in hippocampal neurons upon current ramps under no light, continuous or 10 Hz pulsed illumination with 550 or 635 nm light.
Appendix 1—figure 7
Soma targeting of MsACR1 and raACR.
(A) Stimulation and illumination paradigms used to assess silencing efficiency for soma-targeted MsACR1 and raACR in dissociated hippocampal neurons. (B) Spikes elicited by current-ramp injection in …
Appendix 1—figure 8
Modeling light propagation at 635 and 550 nm via Monte Carlo simulation.
To model the propagation of photons with different wavelengths (635 and 550 nm) through brain tissue, we utilized the script developed by Stujenske et al., 2015. We initialized the script with a …
Appendix 1—figure 9
Activation of MsACR1 and raACR in PV-positive neurons in primary motor cortex leads to reduced locomotion in OpenField test.
(A) Speed and trajectory of one example raACR mouse across five stimulation bouts (red highlights). (B) Calculated average velocity 2 s prior to and during light stimulation. Each mouse underwent …
Additional files
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MDAR checklist
- https://cdn.elifesciences.org/articles/90100/elife-90100-mdarchecklist1-v1.docx
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Appendix 1—figure 3—source data 1
Source data for electrophysiological analysis to determine light sensitivity and ion selectivity of identified ACRs presented in Appendix 1—figure 3A-C, E, F.
- https://cdn.elifesciences.org/articles/90100/elife-90100-app1-fig3-data1-v1.xlsx
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Appendix 1—figure 5—source data 1
Source data for electrophysiological analysis to determine red-light activity of ACRs presented in Appendix 1—figure 5B and C.
- https://cdn.elifesciences.org/articles/90100/elife-90100-app1-fig5-data1-v1.xlsx
