Optical control of zebrafish behavior with halorhodopsin. Proc Natl Acad Sci USA

Department of Physiology, Program in Neuroscience, University of California, San Francisco, 1550 4th Street, San Francisco, CA 94158-2324, USA.
Proceedings of the National Academy of Sciences (Impact Factor: 9.67). 10/2009; 106(42):17968-73. DOI: 10.1073/pnas.0906252106
Source: PubMed


Expression of halorhodopsin (NpHR), a light-driven microbial chloride pump, enables optical control of membrane potential and reversible silencing of targeted neurons. We generated transgenic zebrafish expressing enhanced NpHR under control of the Gal4/UAS system. Electrophysiological recordings showed that eNpHR stimulation effectively suppressed spiking of single neurons in vivo. Applying light through thin optic fibers positioned above the head of a semi-restrained zebrafish larva enabled us to target groups of neurons and to simultaneously test the effect of their silencing on behavior. The photostimulated volume of the zebrafish brain could be marked by subsequent photoconversion of co-expressed Kaede or Dendra. These techniques were used to localize swim command circuitry to a small hindbrain region, just rostral to the commissura infima Halleri. The kinetics of the hindbrain-generated swim command was investigated by combined and separate photo-activation of NpHR and Channelrhodopsin-2 (ChR2), a light-gated cation channel, in the same neurons. Together this "optogenetic toolkit" allows loss-of-function and gain-of-function analyses of neural circuitry at high spatial and temporal resolution in a behaving vertebrate.

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    • "We have previously observed that inhibition with NpHR and NpHR2.0, while useful for many applications (Gradinaru et al., 2009; Sohal et al., 2009; Tønnesen et al., 2009; Arrenberg et al., 2009), can in some cases be overcome by very strong excitation (Sohal et al., 2009). Hyperpolarizations by greater than 100 mV (Figure 1) with eNpHR3.0 "
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    DESCRIPTION: Molecular and Cellular Approaches for Diversifying and Extending Optogenetics (Viviana Gradinaru,Feng Zhang) university of california , los anjeles and caltech
    • "More recently, Jaws derived from Haloarcula salinarum strain Shark, was demonstrated to be activated by the most red-shifted spectrum of any hyperpolarizing rhodopsin so far (Chuong et al., 2014). Many functions of neurons or neural circuits have been revealed by hyperpolarizing optogenetic rhodopsins (Tønnesen et al., 2009; Arrenberg et al., 2009; Leifer et al., 2011; Tye et al., 2011; Gentet et al., 2012). To apply optogenetics, three requirements are necessary to be fulfilled; the selection of optogenetic actuators, the targeted expression of optogenetic actuators in the neurons or regions of interest and the light delivery system (Yawo et al., 2013). "
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    ABSTRACT: Epilepsy is a neurological disorder that affects around 1% of the population worldwide. The two main therapies, pharmacology and the electrical stimulation, both have some shortcomings. For instance, pharmacological therapy is frequently accompanied by side effects, and current anticonvulsive drugs fail to be effective to around a third of patients. These patients could suffer astrocyte-related epilepsy, as increasing evidence indicates that dysfunctions of astrocytes can result in epilepsy. However, epilepsy drugs that affect astrocytes are not available currently. Although electrical stimulation has benefited many patients, the electrode stimulates unselective neurons or circuits. All these need to develop new strategies for improving the life of the patients. As channelrhodopsins (ChRs) were discovered, a novel method referred to as “optogenetics” was developed. It has advantages over electrical stimulation of being less-invasiveness and allowing spatiotemporally stimulation. Recently, a number of experiments have explored the treatments for epilepsy with optogenetic control of neurons. Here, we discuss the possibility that an optogenetic approach could be used to control the release of gliotransmitters and improve astrocyte function such as glutamate and K+ uptake, and thereby offer a potential strategy to investigate and treat astrocyte-related epilepsy.
    Brain Research Bulletin 11/2014; 110. DOI:10.1016/j.brainresbull.2014.10.013 · 2.72 Impact Factor
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    • "Genes important for zebrafish brain development have been identified in large scale mutagenesis screens (Driever et al. 1996; Granato et al. 1996; Haffter & Nusslein-Volhard 1996; Schier et al. 1996) and can be knocked down by injecting antisense morpholinos into the developing embryo (Heasman 2002). Optogenetic systems can be used to visualize not only anatomy but also neuronal activity during development and behavior in transgenic zebrafish that have been designed to express a genetically encoded calcium sensor in their cells that fluoresces whenever the cells become active (Arrenberg, Del Bene & Baier 2009; Del Bene & Wyart 2012; Higashijima et al. 2003). Finally, any delayed deleterious effects of early life stage toxicant exposure on reproduction, sexual and social behavior, motivation, and cognition can be examined in the adult zebrafish 3-4 months post-fertilization. "
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    ABSTRACT: This chapter highlights current research using the embryonic zebrafish as a model system to study the effects of environmental toxicants on behavior. Zebrafish are ideally suited for high-throughput analyses of behavior. Hundreds of fertilized eggs can be collected daily from a single tank and the synchronously developing embryos can be exposed to environmental toxicants in a culture dish or multiwell plate. The developing motor network of the zebrafish embryo has been impressively characterized at multiple levels, from behavior to circuitry to genes, thus establishing a solid foundation for investigating mechanisms of neurobehavioral toxicity. Three assays are reviewed and their usage to screen for toxicant-induced behavioral defects in embryonic zebrafish is critically evaluated. The mechanisms of neurodevelopment are well conserved in vertebrate species in that similar genes, neurotransmitters, and hormones control early brain development and behavior in fish, mice, and humans. Consequently, behavioral assays in embryonic zebrafish may be used to screen for environmental toxicants that influence human brain development and behavior. Several recommendations are made to strengthen current approaches to accomplishing this important goal.
    Zebrafish, Edited by Charles Lessman, Ethan Carver, 04/2014: chapter 12: pages 245-264; Nova., ISBN: 978-1-63117-558-9
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