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Optogenetic Approaches in Neuroscience

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A special issue of International Journal of Molecular Sciences (ISSN 1422-0067). This special issue belongs to the section "Biochemistry".

Deadline for manuscript submissions: closed (31 October 2017) | Viewed by 43346

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Special Issue Editor

Prof. Dr. Thomas Knöpfel

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Guest Editor

1. Laboratory for Neuronal Circuit Dynamics, Imperial College London, London W12 0NN, UK
2. Centre for Neurotechnology, Institute of Biomedical Engineering, Imperial College London, London SW7 2AZ, UK
Interests: optogenetics; neuronal circuits; optogenetic tools; optogenetic approaches; serotonin
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

Optogenetics can be defined as concepts and methods that are based on the use of light and genetics to manipulate and monitor the activities of defined cell populations. Optogenetic approches already had a major impact in neurosciences, yet are still evolving.

We invite you to contribute original articles that describe conceptual and methodological advances associated with optogenetic approaches. Review articles describing cutting edge and emerging optogenetic technologies are also welcome.

Thomas Knöpfel
Guest Editor

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Keywords

Optogenetic tools
Targeting cell classes
Targeting subcellular structures
Genetically encoded indicators
Genetically encoded actuators
Optogenetic interference in animal models of diseases

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Published Papers (7 papers)

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Research

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5457 KiB

Open AccessArticle

Frequency-Dependent Multi-Well Cardiotoxicity Screening Enabled by Optogenetic Stimulation

by Susanne Rehnelt, Daniela Malan, Krisztina Juhasz, Benjamin Wolters, Leo Doerr, Matthias Beckler, Ralf Kettenhofen, Heribert Bohlen, Tobias Bruegmann and Philipp Sasse

Int. J. Mol. Sci. 2017, 18(12), 2634; https://doi.org/10.3390/ijms18122634 - 6 Dec 2017

Cited by 24 | Viewed by 6678

Abstract

Side effects on cardiac ion channels causing lethal arrhythmias are one major reason for drug withdrawals from the market. Field potential (FP) recording from cardiomyocytes, is a well-suited tool to assess such cardiotoxic effects of drug candidates in preclinical drug development, but it is currently limited to the spontaneous beating of the cardiomyocytes and manual analysis. Herein, we present a novel optogenetic cardiotoxicity screening system suited for the parallel automated frequency-dependent analysis of drug effects on FP recorded from human pluripotent stem cell-derived cardiomyocytes. For the expression of the light-sensitive cation channel Channelrhodopsin-2, we optimised protocols using virus transduction or transient mRNA transfection. Optical stimulation was performed with a new light-emitting diode lid for a 96-well FP recording system. This enabled reliable pacing at physiologically relevant heart rates and robust recording of FP. Thereby we detected rate-dependent effects of drugs on Na⁺, Ca²⁺ and K⁺ channel function indicated by FP prolongation, FP shortening and the slowing of the FP downstroke component, as well as generation of afterdepolarisations. Taken together, we present a scalable approach for preclinical frequency-dependent screening of drug effects on cardiac electrophysiology. Importantly, we show that the recording and analysis can be fully automated and the technology is readily available using commercial products. Full article

(This article belongs to the Special Issue Optogenetic Approaches in Neuroscience)

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7372 KiB

Open AccessArticle

Absorption and Emission Spectroscopic Investigation of Thermal Dynamics and Photo-Dynamics of the Rhodopsin Domain of the Rhodopsin-Guanylyl Cyclase from the Nematophagous Fungus Catenaria anguillulae

by Alfons Penzkofer, Ulrike Scheib, Katja Stehfest and Peter Hegemann

Int. J. Mol. Sci. 2017, 18(10), 2099; https://doi.org/10.3390/ijms18102099 - 5 Oct 2017

Cited by 6 | Viewed by 4938

Abstract

The rhodopsin-guanylyl cyclase from the nematophagous fungus Catenaria anguillulae belongs to a recently discovered class of enzymerhodopsins and may find application as a tool in optogenetics. Here the rhodopsin domain CaRh of the rhodopsin-guanylyl cyclase from Catenaria anguillulae was studied by absorption and emission spectroscopic methods. The absorption cross-section spectrum and excitation wavelength dependent fluorescence quantum distributions of CaRh samples were determined (first absorption band in the green spectral region). The thermal stability of CaRh was studied by long-time attenuation measurements at room temperature (20.5 °C) and refrigerator temperature of 3.5 °C. The apparent melting temperature of CaRh was determined by stepwise sample heating up and cooling down (obtained apparent melting temperature: 62 ± 2 °C). The photocycle dynamics of CaRh was investigated by sample excitation to the first inhomogeneous absorption band of the CaRh_d_a dark-adapted state around 590 nm (long-wavelength tail), 530 nm (central region) and 470 nm (short-wavelength tail) and following the absorption spectra development during exposure and after exposure (time resolution 0.0125 s). The original protonated retinal Schiff base PRSB_all_-trans in CaRh_da photo-converted reversibly to protonated retinal Schiff base PRSB_all_-trans_,_la1 with restructured surroundings (CaRh_la₁ light-adapted state, slightly blue-shifted and broadened first absorption band, recovery to CaRh_da with time constant of 0.8 s) and deprotonated retinal Schiff base RSB₁₃_-cis (CaRh_la₂ light-adapted state, first absorption band in violet to near ultraviolet spectral region, recovery to CaRh_da with time constant of 0.35 s). Long-time light exposure of light-adapted CaRh_la₁ around 590, 530 and 470 nm caused low-efficient irreversible degradation to photoproducts CaRh_pr_od. Schemes of the primary photocycle dynamics of CaRh_da and the secondary photocycle dynamics of CaRh_la1 are developed. Full article

(This article belongs to the Special Issue Optogenetic Approaches in Neuroscience)

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8958 KiB

Open AccessArticle

Transgenic Strategies for Sparse but Strong Expression of Genetically Encoded Voltage and Calcium Indicators

by Chenchen Song, Quyen B. Do, Srdjan D. Antic and Thomas Knöpfel

Int. J. Mol. Sci. 2017, 18(7), 1461; https://doi.org/10.3390/ijms18071461 - 7 Jul 2017

Cited by 19 | Viewed by 6162

Abstract

Rapidly progressing development of optogenetic tools, particularly genetically encoded optical indicators, enables monitoring activities of neuronal circuits of identified cell populations in longitudinal in vivo studies. Recently developed advanced transgenic approaches achieve high levels of indicator expression. However, targeting non-sparse cell populations leads to dense expression patterns such that optical signals from neuronal processes cannot be allocated to individual neurons. This issue is particularly pertinent for the use of genetically encoded voltage indicators whose membrane-delimited signals arise largely from the neuropil where dendritic and axonal membranes of many cells intermingle. Here we address this need for sparse but strong expression of genetically encoded optical indicators using a titratable recombination-activated transgene transcription to achieve a Golgi staining-type indicator expression pattern in vivo. Using different transgenic strategies, we also illustrate that co-expression of genetically encoded voltage and calcium indicators can be achieved in vivo for studying neuronal circuit input–output relationships. Full article

(This article belongs to the Special Issue Optogenetic Approaches in Neuroscience)

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Review

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636 KiB

Open AccessReview

Optogenetic Investigation of Arousal Circuits

by Susan M. Tyree and Luis De Lecea

Int. J. Mol. Sci. 2017, 18(8), 1773; https://doi.org/10.3390/ijms18081773 - 15 Aug 2017

Cited by 23 | Viewed by 6131

Abstract

Modulation between sleep and wake states is controlled by a number of heterogeneous neuron populations. Due to the topological proximity and genetic co-localization of the neurons underlying sleep-wake state modulation optogenetic methods offer a significant improvement in the ability to benefit from both the precision of genetic targeting and millisecond temporal control. Beginning with an overview of the neuron populations mediating arousal, this review outlines the progress that has been made in the investigation of arousal circuits since the incorporation of optogenetic techniques and the first in vivo application of optogenetic stimulation in hypocretin neurons in the lateral hypothalamus. This overview is followed by a discussion of the future progress that can be made by incorporating more recent technological developments into the research of neural circuits. Full article

(This article belongs to the Special Issue Optogenetic Approaches in Neuroscience)

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2471 KiB

Open AccessReview

Recent Progress of Development of Optogenetic Implantable Neural Probes

by Hubin Zhao

Int. J. Mol. Sci. 2017, 18(8), 1751; https://doi.org/10.3390/ijms18081751 - 11 Aug 2017

Cited by 26 | Viewed by 8535

Abstract

As a cell type-specific neuromodulation method, optogenetic technique holds remarkable potential for the realisation of advanced neuroprostheses. By genetically expressing light-sensitive proteins such as channelrhodopsin-2 (ChR2) in cell membranes, targeted neurons could be controlled by light. This new neuromodulation technique could then be applied into extensive brain networks and be utilised to provide effective therapies for neurological disorders. However, the development of novel optogenetic implants is still a key challenge in the field. The major requirements include small device dimensions, suitable spatial resolution, high safety, and strong controllability. In this paper, I present a concise review of the significant progress that has been made towards achieving a miniaturised, multifunctional, intelligent optogenetic implant. I identify the key limitations of current technologies and discuss the possible opportunities for future development. Full article

(This article belongs to the Special Issue Optogenetic Approaches in Neuroscience)

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3788 KiB

Open AccessReview

Bacterial Phytochromes, Cyanobacteriochromes and Allophycocyanins as a Source of Near-Infrared Fluorescent Probes

by Olena S. Oliinyk, Konstantin G. Chernov and Vladislav V. Verkhusha

Int. J. Mol. Sci. 2017, 18(8), 1691; https://doi.org/10.3390/ijms18081691 - 3 Aug 2017

Cited by 49 | Viewed by 8422

Abstract

Bacterial photoreceptors absorb light energy and transform it into intracellular signals that regulate metabolism. Bacterial phytochrome photoreceptors (BphPs), some cyanobacteriochromes (CBCRs) and allophycocyanins (APCs) possess the near-infrared (NIR) absorbance spectra that make them promising molecular templates to design NIR fluorescent proteins (FPs) and biosensors for studies in mammalian cells and whole animals. Here, we review structures, photochemical properties and molecular functions of several families of bacterial photoreceptors. We next analyze molecular evolution approaches to develop NIR FPs and biosensors. We then discuss phenotypes of current BphP-based NIR FPs and compare them with FPs derived from CBCRs and APCs. Lastly, we overview imaging applications of NIR FPs in live cells and in vivo. Our review provides guidelines for selection of existing NIR FPs, as well as engineering approaches to develop NIR FPs from the novel natural templates such as CBCRs. Full article

(This article belongs to the Special Issue Optogenetic Approaches in Neuroscience)

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