Targeting and Readout Strategies for Fast Optical Neural Control<i>In Vitro</i>and<i>In Vivo</i>

Gradinaru, Viviana; Thompson, Kimberly R.; Zhang, Feng; Mogri, Murtaza; Kay, Kenneth; Schneider, M. Bret; Deisseroth, Karl

doi:10.1523/jneurosci.3578-07.2007

Cited by 443 publications

(426 citation statements)

References 67 publications

(98 reference statements)

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“…Optrode recording was performed in isofluraneanesthetized mice on a stereotaxic frame (KOPF), and recording was carried out as described previously (51). Recordings were performed on stroke mice at poststroke day 5.…”

Section: Methodsmentioning

confidence: 99%

Optogenetic neuronal stimulation promotes functional recovery after stroke

Cheng

Wang

Woodson

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

182

205

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Clinical and research efforts have focused on promoting functional recovery after stroke. Brain stimulation strategies are particularly promising because they allow direct manipulation of the target area's excitability. However, elucidating the cell type and mechanisms mediating recovery has been difficult because existing stimulation techniques nonspecifically target all cell types near the stimulated site. To circumvent these barriers, we used optogenetics to selectively activate neurons that express channelrhodopsin 2 and demonstrated that selective neuronal stimulations in the ipsilesional primary motor cortex (iM1) can promote functional recovery. Stroke mice that received repeated neuronal stimulations exhibited significant improvement in cerebral blood flow and the neurovascular coupling response, as well as increased expression of activity-dependent neurotrophins in the contralesional cortex, including brain-derived neurotrophic factor, nerve growth factor, and neurotrophin 3. Western analysis also indicated that stimulated mice exhibited a significant increase in the expression of a plasticity marker growth-associated protein 43. Moreover, iM1 neuronal stimulations promoted functional recovery, as stimulated stroke mice showed faster weight gain and performed significantly better in sensory-motor behavior tests. Interestingly, stimulations in normal nonstroke mice did not alter motor behavior or neurotrophin expression, suggesting that the prorecovery effect of selective neuronal stimulations is dependent on the poststroke environment. These results demonstrate that stimulation of neurons in the stroke hemisphere is sufficient to promote recovery. stroke recovery | channelrhodopsin

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Section: Methodsmentioning

confidence: 99%

Optogenetic neuronal stimulation promotes functional recovery after stroke

Cheng

Wang

Woodson

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

182

205

View full text Add to dashboard Cite

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“…Given the potential for side effects of expressing opsins in cells, assessing toxicity may be of utility for specific scientific or preclinical questions, and can be aided by analyzing cells with respect to accepted criteria or markers of toxicity for the cell type of interest (Gradinaru et al 2007;Han et al 2009;Doroudchi et al 2011;Miyashita et al 2013). Experiments to assess toxicity ideally should insure that an opsin (or its attached fluorophore) has not adversely altered gross cell morphology or function, and may include histology, electrical recordings, molecular profiling, and other strategies.…”

Section: Cell-autonomous Side Effects Protein Expressionmentioning

confidence: 99%

Principles of designing interpretable optogenetic behavior experiments

2015

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Over the last decade, there has been much excitement about the use of optogenetic tools to test whether specific cells, regions, and projection pathways are necessary or sufficient for initiating, sustaining, or altering behavior. However, the use of such tools can result in side effects that can complicate experimental design or interpretation. The presence of optogenetic proteins in cells, the effects of heat and light, and the activity of specific ions conducted by optogenetic proteins can result in cellular side effects. At the network level, activation or silencing of defined neural populations can alter the physiology of local or distant circuits, sometimes in undesired ways. We discuss how, in order to design interpretable behavioral experiments using optogenetics, one can understand, and control for, these potential confounds.

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“…13 ). The substantially red-shifted action spectrum will allow not only combinatorial control experiments, but also improved integration with existing blue lightactivated Ca 2+ indicators and deeper penetration of light with in vivo applications 14 as a result of the reduced scattering of lower-energy photons.…”

mentioning

confidence: 99%

Red-shifted optogenetic excitation: a tool for fast neural control derived from Volvox carteri

et al. 2008

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The introduction of two microbial opsin-based tools, channelrhodopsin-2 (ChR2) and halorhodopsin (NpHR), to neuroscience has generated interest in fast, multimodal, cell type-specific neural circuit control. Here we describe a cation-conducting channelrhodopsin (VChR1) from Volvox carteri that can drive spiking at 589 nm, with excitation maximum red-shifted ~70 nm compared with ChR2. These results demonstrate fast photostimulation with yellow light, thereby defining a functionally distinct third category of microbial rhodopsin proteins.Microbial proteins that can be rapidly activated by light have been adapted for research in neuroscience, including ChR2 and NpHR, which permit millisecond-precision optical control of genetically defined cell types in intact neural tissue 1-6 . Because ChR2 is a blue light-gated cation channel and NpHR is a yellow light-driven chloride pump, the combination of these two proteins allows independent neural excitation and inhibition in the same preparation. However, there has been enormous interest in developing a hypothetical third major optogenetic tool, namely a second cation channel with an action spectrum that is substantially red-shifted relative to ChR2, to allow tests of the interaction of cell types in circuit computation or behavior.Although efforts to develop a distinct light-activated excitatory protein have been focused on molecular engineering of ChR2, another approach would be to identify previously unknown microbial channelrhodopsins using genomic tools. One ChR2-related sequence from the spheroidal alga Volvox carteri (Fig. 1a) has been described, but the absorption spectrum of the protein and the photocycle dynamics are virtually identical to those of ChR2 (refs. 7 ,8 ). Therefore, we searched the genome database from the US Department of Energy Joint Genome Institute, discovered a second Volvox ChR (VChR1) that is more related to ChR1 (ref. 9) from Chlamydomonas reinhardtii, explored its properties in heterologous expression systems and functionally tested the codon-optimized opsin gene in mammalian neurons.We expressed VChR1 in Xenopus oocytes and HEK293 cells and observed evoked photocurrents similar to those of ChR1 from Chlamydomonas 9,10 . The photocurrents were graded with light intensity and showed inactivation from a fast peak toward a reduced stationary plateau (Fig. 1b) (Fig. 1b) 9 , and currents showed an inwardly rectifying current-voltage relationship (Fig. 1c).Certain primary structural differences between VChR1 and the Chlamydomonas ChRs suggested that the properties of VChR1 would be distinct from those of the other ChRs (Fig. 1d). On the basis of electrostatic potential and quantum mechanical-molecular mechanical calculations for bacteriorhodopsin and relatives, the counterion complex of the all-trans retinal Schiff base (RSB; Fig. 1d) should be critical for color tuning 11,12 , but these residues are conserved in both ChR1 and VChR1 (blue sequence, Fig. 1d). On the other hand, calculations and mutational experiments 11,12 predict that fo...

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Targeting and Readout Strategies for Fast Optical Neural ControlIn VitroandIn Vivo

Cited by 443 publications

References 67 publications

Optogenetic neuronal stimulation promotes functional recovery after stroke

Optogenetic neuronal stimulation promotes functional recovery after stroke

Principles of designing interpretable optogenetic behavior experiments

Red-shifted optogenetic excitation: a tool for fast neural control derived from Volvox carteri

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