Channelrhodopsins are retinylidene proteins unique in their ability to generate light-gated passive ion currents across the membrane. Heterologous expression of channelrhodopsins in animal cells, most notably, neurons, enables precise millisecond-scale control of their activity (optogenetics). For 15 years only cation-conducting channelrhodopsins derived from chlorophyte (green) algae have been known, which now are widely used in neuroscience research to stimulate neurons with light. Recently a distinct family of channelrhodopsins with strictly anion selectivity has been discovered in phylogenetically distant cryptophyte flagellate algae. Furthermore, the genomes of the latter microorganisms also encode a structurally distinct group of rhodopsins capable of cation channel activity using different molecular mechanisms as compared to their chlorophyte counterparts. In this review, we mostly focus on the two latest additions and their utility for optogenetic applications.
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References
Alfonsa H., Lakey J. H., Lightowlers R. N., and Trevelyan A. J., “Cl-out is a novel cooperative optogenetic tool for extruding chloride from neurons,” Nat. Commun., 7, 13495 (2016).
Andrasfalvy B. K., Zemelman B. V., Tang J., and Vaziri A., “Two-photon single-cell optogenetic control of neuronal activity by sculpted light,” Proc. Natl. Acad. Sci. USA, 107, 11981–11986 (2010).
Bamann C., Gueta R., Kleinlogel S., et al., “Structural guidance of the photocycle of channelrhodopsin-2 by an interhelical hydrogen bond,” Biochemistry, 49, 267–278 (2010).
Berndt A., Lee S. Y., Wietek J., et al., “Structural foundations of optogenetics: Determinants of channelrhodopsin ion selectivity,” Proc. Natl. Acad. Sci. USA, 113, 822–829 (2016).
Berndt A., Yizhar O., Gunaydin L. A., et al., “Bi-stable neural state switches,” Nat. Neurosci., 12, 229–234 (2009).
Boyden E. S., “Optogenetics and the future of neuroscience,” Nat. Neurosci., 18, 1200–1201 (2015).
Boyden E. S., Zhang F., Bamberg E., et al., “Millisecond-timescale, genetically targeted optical control of neural activity,” Nat. Neurosci., 8, 1263–1268 (2005).
Chow B. Y. and Boyden E. S., “Optogenetics and translational medicine,” Sci. Transl. Med., 5, 177ps175 (2013).
Cohen A. E., “Optogenetics: Turning the microscope on its head,” Biophys. J., 110, 997–1003 (2016).
Deisseroth K., “Optogenetics: 10 years of microbial opsins in neuroscience,” Nat. Neurosci., 18, 1213–1225 (2015).
Deisseroth K., Feng G., Majewska A. K., et al., “Next-generation optical technologies for illuminating genetically targeted brain circuits,” J. Neurosci., 26, 10380–10386 (2006).
Dolgikh D. A., Malyshev A. Yu., Roshchin M. V., et al., “Comparative characteristics of two anion-channel rhodopsins and prospects of their use in optogenetics,” Dokl. Biochem. Biophys., 471, 440–442 (2016).
Feldbauer K., Zimmermann D., Pintschovius V., et al., “Channelrhodopsin-2 is a leaky proton pump,” Proc. Natl. Acad. Sci. USA, 106, 12317–12322 (2009).
Foster K.-W., Saranak J., Patel N., et al., “A rhodopsin is the functional photoreceptor for phototaxis in the unicelullar eukaryote Chlamydomonas,” Nature, 311, 756–759 (1984).
Govorunova E. G., Cunha C. R., Sineshchekov O. A., and Spudich J. L. “Anion channelrhodopsins for inhibitory cardiac optogenetics,” Sci. Rep., 6, 33530 (2016).
Govorunova E. G., Sineshchekov O. A., Li H., et al., “Characterization of a highly efficient blue-shifted channelrhodopsin from the marine alga Platymonas subcordiformis,” J. Biol. Chem., 288, 29911–29922 (2013).
Govorunova E. G., Sineshchekov O. A., Li H., and Spudich J. L. “Microbial rhodopsins: Diversity, mechanisms, and optogenetic applications,” Annu. Rev. Biochem. (2017а).
Govorunova E. G., Sineshchekov O. A., Liu X., et al., “Natural light-gated anion channels: A family of microbial rhodopsins for advanced optogenetics,” Science, 349, 647–650 (2015).
Govorunova E. G., Sineshchekov O. A., Rodarte E. M., et al., “The expanding family of natural anion channelrhodopsins reveals large variations in kinetics, conductance, and spectral sensitivity,” Sci. Rep., 7, 43358 (2017b).
Govorunova E. G., Sineshchekov O. A., and Spudich J. L. “Structurally distinct cation channelrhodopsins from cryptophyte algae,” Biophys. J., 110, 2302–2304 (2016a).
Hou S. Y., Govorunova E. G., Ntefidou M., et al., “Diversity of Chlamydomonas channelrhodopsins,” Photochem. Photobiol., 88, 119–128 (2012).
Kato H. E., Kamiya M., Sugo S., et al., “Atomistic design of microbial opsin-based blue-shifted optogenetics tools,” Nat. Commun., 6, 7177 (2015).
Kato H. E., Zhang F., Yizhar O., et al., “Crystal structure of the channelrhodopsin light-gated cation channel,” Nature, 482, 369–374 (2012).
Keeling P. J., Burki F., Wilcox H. M., et al., “The Marine Microbial Eukaryote Transcriptome Sequencing Project (MMETSP): illuminating the functional diversity of eukaryotic life in the oceans through transcriptome sequencing,” PLoS Biol., 12, e1001889 (2014).
Klapoetke N. C., Murata Y., Kim S. S., et al., “Independent optical excitation of distinct neural populations,” Nat. Methods, 11, 338–346 (2014).
Klapper S. D., Swiersy A., Bamberg E., and Busskamp V., “Biophysical properties of optogenetic tools and their application for vision restoration approaches,” Front. Syst. Neurosci., 10, 74 (2016).
Kleinlogel S., Feldbauer K., Dempski R. E., et al., “Ultra light-sensitive and fast neuronal activation with the Ca2+-permeable channelrhodopsin CatCh,” Nat. Neurosci., 14, 513–518 (2011).
Lewis T. L., Jr., Mao T., Svoboda K., and Arnold D. B., “Myosin-dependent targeting of transmembrane proteins to neuronal dendrites,” Nat. Neurosci., 12, 568–576 (2009).
Li X., Gutierrez D. V., Hanson M. G., et al., “Fast noninvasive activation and inhibition of neural and network activity by vertebrate rhodopsin and green algae channelrhodopsin,” Proc. Natl. Acad. Sci. USA, 102, 17816–17821 (2005).
Lin J. Y., Knutsen P. M., Muller A., et al., “ReaChR: a red-shifted variant of channelrhodopsin enables deep transcranial optogenetic excitation,” Nat. Neurosci., 16, 1499–1508 (2013).
Lin J. Y., Lin M. Z., Steinbach P., and Tsien R. Y., “Characterization of engineered channelrhodopsin variants with improved properties and kinetics,” Biophys. J., 96, 1803–1814 (2009).
Litvin F. F., Sineshchekov O. A., and Sineshchekov V. A., “Photoreceptor electric potential in the phototaxis of the alga Haematococcus pluvialis,” Nature, 271, 476–478 (1978).
Lorenz-Fonfria V. A. and Heberle J., “Channelrhodopsin unchained: structure and mechanism of a light-gated cation channel,” Biochim. Biophys. Acta, 1837, 626–642 (2014).
Lorenz-Fonfria V. A., Resler T., Krause N., et al., “Transient protonation changes in channelrhodopsin-2 and their relevance to channel gating,” Proc. Natl. Acad. Sci. USA, 110, E1273–1281 (2013).
Lorincz M. L. and Adamantidis A. R., “Monoaminergic control of brain states and sensory processing: Existing knowledge and recent insights obtained with optogenetics,” Prog. Neurobiol., 151, 237–253 (2017).
Mahn M., Prigge M., Ron S., et al., “Biophysical constraints of optogenetic inhibition at presynaptic terminals,” Nat. Neurosci., 19, 554–556 (2016).
Malyshev A. Y., Roshchin M. V., Smirnova G. R., et al., “Chloride conducting light activated channel GtACR2 can produce both cessation of firing and generation of action potentials in cortical neurons in response to light,” Neurosci. Lett., 640, 76–80 (2017).
Matasci N., Hung L. H., Yan Z., et al., “Data access for the 1,000 Plants (1KP) project,” Gigascience, 3, 17 (2014).
Mohammad F., Stewart J. C., Ott S., et al., “Optogenetic inhibition of behavior with anion channelrhodopsins,” Nat. Methods, 14, 271–274 (2017).
Mukohata Y. and Kaji Y., “Light-induced membrane-potential increase, ATP synthesis, and proton uptake in Halobacterium halobium, R1mR catalyzed by halorhodopsin: Effects of N,N’-dicyclohexylcarbodiimide, triphenyltin chloride, and 3,5-di-tert-butyl-4-hydroxybenzylidenemalononitrile (SF6847),” Arch. Biochem. Biophys., 206, 72–76 (1981).
Nagel G., Brauner M., Liewald J. F., et al., “Light activation of channelrhodopsin-2 in excitable cells of Caenorhabditis elegans triggers rapid behavioral responses,” Curr. Biol., 15, 2279–2284 (2005).
Nagel G., Ollig D., Fuhrmann M., et al., “Channelrhodopsin-1: a light-gated proton channel in green algae,” Science, 296, 2395–2398 (2002).
Nagel G., Szellas T., Huhn W., et al., “Channelrhodopsin-2, a directly light-gated cation-selective membrane channel,” Proc. Natl. Acad. Sci. USA, 100, 13940–13945 (2003).
Oesterhelt D. and Stoeckenius W., “Rhodopsin-like protein from the purple membrane of Halobacterium halobium,” Nature, 233, 149–152 (1971).
Park S. A., Lee S. R., Tung L., and Yue D. T., “Optical mapping of optogenetically shaped cardiac action potentials,” Sci. Rep., 4, 6125 (2014).
Paz J. T., Davidson T. J., Frechette E. S., et al., “Closed-loop optogenetic control of thalamus as a tool for interrupting seizures after cortical injury,” Nat. Neurosci., 16, 64–70 (2013).
Price G. D. and Trussell L. O., “Estimate of the chloride concentration in a central glutamatergic terminal: a gramicidin perforated-patch study on the calyx of Held,” J. Neurosci., 26, 11432–11436 (2006).
Schneider F., Grimm C., and Hegemann P., “Biophysics of channelrhodopsin,” Annu. Rev. Biophys., 44, 167–186 (2015).
Schobert B. and Lanyi J. K., “Halorhodopsin is a light-driven chloride pump,” J. Biol. Chem., 257, 10306–10313 (1982).
Sineshchekov O. A., Govorunova E. G., Li H., and Spudich J. L., “Gating mechanisms of a natural anion channelrhodopsin,” Proc. Natl. Acad. Sci. USA, 112, 14236–14241 (2015).
Sineshchekov O. A., Govorunova E. G., and Spudich J. L., “Photosensory functions of channelrhodopsins in native algal cells,” Photochem. Photobiol., 85, 556–563 (2009).
Sineshchekov O. A., Jung K.-H., and Spudich J. L., “Two rhodopsins mediate phototaxis to low- and high-intensity light in Chlamydomonas reinhardtii,” Proc. Natl. Acad. Sci. USA, 99, 8689–8694 (2002).
Sineshchekov O. A., Li H., Govorunova E. G., and Spudich J. L., “Photochemical reaction cycle transitions during anion channelrhodopsin gating,” Proc. Natl. Acad. Sci. USA, 113, E1993–2000 (2016).
Sineshchekov O. A. and Spudich J. L., Sensory Rhodopsin Signaling in Green Flagellate Algae. Handbook of Photosensory Receptors, Wiley-VCH, Weinheim (2005), pp. 25–42.
Tsunoda S. P. and Hegemann P., “Glu 87 of channelrhodopsin-1 causes pH-dependent color tuning and fast photocurrent inactivation,” Photochem. Photobiol., 85, 564–569 (2009).
Verhoefen M. K., Bamann C., Blocher R., et al., “The photocycle of channelrhodopsin-2: Ultrafast reaction dynamics and subsequent reaction steps,” Chemphyschem., 11, 3113–3122 (2010).
Wietek J., Beltramo R., Scanziani M., et al., “An improved chloride-conducting channelrhodopsin for light-induced inhibition of neuronal activity in vivo,” Sci. Rep., 5, 14807 (2015).
Wietek J., Broser M., Krause B. S., and Hegemann P., “Identification of a natural green light absorbing chloride conducting channelrhodopsin from Proteomonas sulcata,” J. Biol. Chem., 291, 4121–4127 (2016).
Wietek J. and Prigge M., “Enhancing channelrhodopsins: An overview,” Methods Mol. Biol., 1408, 141–165 (2016).
Zhang F., Vierock J., Yizhar O., et al., “The microbial opsin family of optogenetic tools,” Cell, 147, 1446–1457 (2011).
Zhang F., Wang L. P., Brauner M., et al., “Multimodal fast optical interrogation of neural circuitry,” Nature, 446, 633–639 (2007).
Zhao M., Alleva R., Ma H., et al., “Optogenetic tools for modulating and probing the epileptic network,” Epilepsy Res., 116, 15–26 (2015).
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Published (in Russian) in Zhurnal Vysshei Nervnoi Deyatel’nosti imeni I. P. Pavlova, Vol. 67, No. 5, pp. 9–17, September–October, 2017.
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Govorunova, E.G., Sineshchekov, О.А. & Spudich, J.L. Three Families of Channelrhodopsins and Their Use in Optogenetics (review). Neurosci Behav Physi 49, 163–168 (2019). https://doi.org/10.1007/s11055-019-00710-6
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DOI: https://doi.org/10.1007/s11055-019-00710-6