Abstract
Channelrhodopsins (ChRs) are the light-gated ion channels that have opened the research field of optogenetics. They were originally identified in the green alga Chlamydomonas reinhardtii, a biciliated unicellular alga that inhabits in freshwater, swims with the cilia, and undergoes photosynthesis. It has various advantages as an experimental organism and is used in a wide range of research fields including photosynthesis, cilia, and sexual reproduction. ChRs function as the primary photoreceptor for the cell’s photo-behavioral responses, seen as changes in the manner of swimming after photoreception. In this chapter, we will introduce C. reinhardtii as an experimental organism and explain our current understanding of how the cell senses light and shows photo-behavioral responses.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Abbreviations
- CGL:
-
Carotenoid granule layers
- ChR:
-
Channelrhodopsin
- Cop:
-
Chlamyopsin
- EST:
-
Expressed sequence tag
- IFT:
-
Intraflagellar transport
- PRC:
-
Photoreceptor current
- PSY:
-
Phytoene synthase
References
Asamizu E, Nakamura Y, Sato S, Fukuzawa H, Tabata S (1999) A large scale structural analysis of cDNAs in a unicellular green alga, Chlamydomonas reinhardtii. I. Generation of 3433 non-redundant expressed sequence tags. DNA Res 6:369–373
Asamizu E et al (2000) Generation of expressed sequence tags from low-CO2 and high-CO2 adapted cells of Chlamydomonas reinhardtii. DNA Res 7:305–307
Awasthi M, Ranjan P, Sharma K, Veetil SK, Kateriya S (2016) The trafficking of bacterial type rhodopsins into the Chlamydomonas eyespot and flagella is IFT mediated. Sci Rep 6:34646
Berthold P, Tsunoda SP, Ernst OP, Mages W, Gradmann D, Hegemann P (2008) Channelrhodopsin-1 initiates phototaxis and photophobic responses in Chlamydomonas by immediate light-induced depolarization. Plant Cell 20:1665–1677
Bessen M, Fay RB, Witman GB (1980) Calcium control of waveform in isolated flagellar axonemes of Chlamydomonas. J Cell Biol 86:446–455
Boyd JS, Lamb MR, Dieckmann CL (2011a) Miniature- and multiple-eyespot loci in Chlamydomonas reinhardtii define new modulators of eyespot photoreception and assembly. G3 (Bethesda) 1:489–498
Boyd JS, Mittelmeier TM, Lamb MR, Dieckmann CL (2011b) Thioredoxin-family protein EYE2 and Ser/Thr kinase EYE3 play interdependent roles in eyespot assembly. Mol Biol Cell 22:1421–1429
Choudhary SK, Baskaran A, Sharma P (2019) Reentrant efficiency of Phototaxis in Chlamydomonas reinhardtii cells. Biophys J 117:1508–1513
Cole DG (2003) The intraflagellar transport machinery of Chlamydomonas reinhardtii. Traffic 4:435–442
Cong L et al (2013) Multiplex genome engineering using CRISPR/Cas systems. Science 339:819–823
Demmig-Adams B, Adams WW 3rd (1992) Photoprotection and other responses of plants to high light stress. Ann Rev Plant Physiol Plant Mol Biol 43:599–626
Dieckmann CL (2003) Eyespot placement and assembly in the green alga Chlamydomonas. Bioessays 25:410–416
Dutcher SK (2003) Elucidation of basal body and centriole functions in Chlamydomonas reinhardtii. Traffic 4:443–451
Eichenberger W, Boschetti A, Michel HP (1986) Lipid and pigment composition of a chlorophyll beta-deficient mutant of Chlamydomonas reinhardii. Physiol Plant 66:589–594
Foster KW, Smyth RD (1980) Light antennas in phototactic algae. Microbiol Rev 44:572–630
Foster KW, Saranak J, Patel N, Zarilli G, Okabe M, Kline T, Nakanishi K (1984) A rhodopsin is the functional photoreceptor for phototaxis in the unicellular eukaryote Chlamydomonas. Nature 311:756–759
Fuhrmann M, Stahlberg A, Govorunova E, Rank S, Hegemann P (2001) The abundant retinal protein of the Chlamydomonas eye is not the photoreceptor for phototaxis and photophobic responses. J Cell Sci 114:3857–3863
Fuhrmann MDW, Kateriya S, Hegemann P (2003) Rhodopsinrelated proteins, cop1, cop2 and chop1, in Chlamydomonas reinhardtii. In: Batschauer A (ed) Photoreceptors and light signaling. Royal Society of Chemistry, Cambridge
Fujiu K, Nakayama Y, Yanagisawa A, Sokabe M, Yoshimura K (2009) Chlamydomonas CAV2 encodes a voltage- dependent calcium channel required for the flagellar waveform conversion. Curr Biol 19:133–139
Gallaher SD, Fitz-Gibbon ST, Glaesener AG, Pellegrini M, Merchant SS (2015) Chlamydomonas genome resource for laboratory strains reveals a mosaic of sequence variation, identifies true strain histories, and enables strain-specific studies. Plant Cell 27:2335–2352
Geimer S, Melkonian M (2004) The ultrastructure of the Chlamydomonas reinhardtii basal apparatus: identification of an early marker of radial asymmetry inherent in the basal body. J Cell Sci 117:2663–2674
Goodenough UW, Weiss RL (1978) Interrelationships between microtubules, a striated fiber, and the gametic mating structure of Chlamydomonas reinhardi. J Cell Biol 76:430–438
Gregonis DE, Rilling HC (1974) The stereochemistry of trans-phytoene synthesis. Some observations on lycopersene as a carotene precursor and a mechanism for the synthesis of cis- and trans-phytoene. Biochemistry 13:1538–1542
Greiner A, Kelterborn S, Evers H, Kreimer G, Sizova I, Hegemann P (2017) Targeting of photoreceptor genes in Chlamydomonas reinhardtii via zinc-finger nucleases and CRISPR/Cas9. Plant Cell 29:2498–2518
Gross CH, Ranum LPW, Lefebvre PA (1988) Extensive restriction fragment length polymorphisms in a new isolate of Chlamydomonas reinhardtii. Curr Genet 13:503–508
Gumpel NJ, Rochaix JD, Purton S (1994) Studies on homologous recombination in the green alga Chlamydomonas reinhardtii. Curr Genet 26:438–442
Harris EH (2009a) Chlamydomonas in the laboratory. In: Harris EH (ed) The Chlamydomonas sourcebook, vol 1, 2nd edn. Academic Press, London, pp 241–302
Harris EH (2009b) The sexual cycle. In: Harris EH (ed) The Chlamydomonas sourcebook, vol 1, 2nd edn. Academic Press, London, pp 119–157
Hartshorne JN (1953) The function of the eyespot in Chlamydomonas. New Phytol 52:292–297
Hayashi M, Yagi T, Yoshimura K, Kamiya R (1998) Real-time observation of Ca2+−induced basal body reorientation in Chlamydomonas. Cell Motil Cytoskeleton 41:49–56
Hegemann P, Berthold P (2009) Sensory photoreceptors and light control of flagellar activity. In: Witman GB (ed) The Chlamydomonas sourcebook, vol 3, 2nd edn. Academic Press, San Diego, CA, pp 395–429
Holmes JA, Dutcher SK (1989) Cellular asymmetry in Chlamydomonas reinhardtii. J Cell Sci 94:273–285
Hyams JS, Borisy GG (1978) Isolated flagellar apparatus of Chlamydomonas: characterization of forward swimming and alteration of waveform and reversal of motion by calcium ions in vitro. J Cell Sci 33:235–253
Ide T, Mochiji S, Ueki N, Yamaguchi K, Shigenobu S, Hirono M, Wakabayashi K (2016) Identification of the agg1 mutation responsible for negative phototaxis in a “wild-type” strain of Chlamydomonas reinhardtii. Biochem Biophys Rep 7:379–385
Iomini C, Li L, Mo W, Dutcher SK, Piperno G (2006) Two flagellar genes, AGG2 and AGG3, mediate orientation to light in Chlamydomonas. Curr Biol 16:1147–1153
Isogai N, Kamiya R, Yoshimura K (2000) Dominance between the two flagella during phototactic turning in Chlamydomonas. Zool Sci 17:1261–1266
Jiang W, Brueggeman AJ, Horken KM, Plucinak TM, Weeks DP (2014) Successful transient expression of Cas9 and single guide RNA genes in Chlamydomonas reinhardtii. Eukaryot Cell 13:1465–1469
Jinek M, Chylinski K, Fonfara I, Hauer M, Doudna JA, Charpentier E (2012) A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity. Science 337:816–821
Kamiya R, Hasegawa E (1987) Intrinsic difference in beat frequency between the two flagella of Chlamydomonas reinhardtii. Exptl Cell Res 173, 299–304
Kamiya R, Witman GB (1984) Submicromolar levels of calcium control the balance of beating between the two flagella in demembranated models of Chlamydomonas. J Cell Biol 98:97–107
Kateriya S, Nagel G, Bamberg E, Hegemann P (2004) “Vision” in single-celled algae. News Physiol Sci 19:133–137
Kong F, Yamaoka Y, Ohama T, Lee Y, Li-Beisson Y (2019) Molecular genetic tools and emerging synthetic biology strategies to increase cellular oil content in Chlamydomonas reinhardtii. Plant Cell Physiol 60:1184–1196
Kozminski KG, Beech PL, Rosenbaum JL (1995) The Chlamydomonas kinesin-like protein FLA10 is involved in motility associated with the flagellar membrane. J Cell Biol 131:1517–1527
Kreimer G (2009) The green algal eyespot apparatus: a primordial visual system and more? Curr Genet 55:19–43
Lamb MR, Dutcher SK, Worley CK, Dieckmann CL (1999) Eyespot-assembly mutants in Chlamydomonas reinhardtii. Genetics 153:721–729
Matsuda A, Yoshimura K, Sineshchekov OA, Hirono M, Kamiya R (1998) Isolation and characterization of novel Chlamydomonas mutants that display phototaxis but not photophobic response. Cell Motil Cytoskeleton 41:353–362
McCarthy SS, Kobayashi MC, Niyogi KK (2004) White mutants of Chlamydomonas reinhardtii are defective in phytoene synthase. Genetics 168:1249–1257
Melis A, Zhang L, Forestier M, Ghirardi ML, Seibert M (2000) Sustained photobiological hydrogen gas production upon reversible inactivation of oxygen evolution in the green alga Chlamydomonas reinhardtii. Plant Physiol 122:127–136
Merchant SS et al (2007) The Chlamydomonas genome reveals the evolution of key animal and plant functions. Science 318:245–250
Mittelmeier TM, Thompson MD, Lamb MR, Lin H, Dieckmann CL (2015) MLT1 links cytoskeletal asymmetry to organelle placement in Chlamydomonas. Cytoskeleton (Hoboken) 72:113–123
Morel-Laurens NML, Feinleib MEH (1983) Photomovement in an “eyeless” mutant of Chlamydomonas. Photochem Photobiol 37:189–194
Nagel G, Ollig D, Fuhrmann M, Kateriya S, Musti AM, Bamberg E, Hegemann P (2002) Channelrhodopsin-1: a light-gated proton channel in green algae. Science 296:2395–2398
Nagel G et al (2003) Channelrhodopsin-2, a directly light-gated cation-selective membrane channel. Proc Natl Acad Sci U S A 100:13940–13945
Nagel G, Szellas T, Kateriya S, Adeishvili N, Hegemann P, Bamberg E (2005) Channelrhodopsins: directly light-gated cation channels. Biochem Soc Trans 33:863–866
Nelson JA, Lefebvre PA (1995) Targeted disruption of the NIT8 gene in Chlamydomonas reinhardtii. Mol Cell Biol 15:5762–5769
Niyogi KK, Bjorkman O, Grossman AR (1997) The roles of specific xanthophylls in photoprotection. Proc Natl Acad Sci U S A 94:14162–14167
Okita N, Isogai N, Hirono M, Kamiya R, Yoshimura K (2005) Phototactic activity in Chlamydomonas 'non-phototactic' mutants deficient in Ca2+-dependent control of flagellar dominance or in inner-arm dynein. J Cell Sci 118:529–537
Pazour GJ, Sineschekov OA, Witman GB (1995) Mutational analysis of the phototransduction pathway of Chlamydomonas reinhardtii. J Cell Biol 131:427–440
Pazour GJ, Dickert BL, Witman GB (1999) The DHC1b (DHC2) isoform of cytoplasmic dynein is required for flagellar assembly. J Cell Biol 144:473–481
Ranum LP, Thompson MD, Schloss JA, Lefebvre PA, Silflow CD (1988) Mapping flagellar genes in Chlamydomonas using restriction fragment length polymorphisms. Genetics 120:109–122
Rüffer U, Nultsch W (1987) Comparison of the beating of cis- and trans-flagella of Chlamydomonas cells held on micropipettes. Cell Motil 7:87–93
Rüffer U, Nultsch W (1991) Flagellar photoresponses of Chlamydomonas cells held on micropipettes: II. Change in flagellar beat pattern. Cell Motil Cytoskeleton 18:269–278
Salisbury JL, Baron AT, Sanders MA (1988) The centrin-based cytoskeleton of Chlamydomonas reinhardtii: distribution in interphase and mitotic cells. J Cell Biol 107:635–641
Sanders MA, Salisbury JL (1989) Centrin-mediated microtubule severing during flagellar excision in Chlamydomonas reinhardtii. J Cell Biol 108:1751–1760
Scranton MA, Ostrand JT, Fields FJ, Mayfield SP (2015) Chlamydomonas as a model for biofuels and bio-products production. Plant J 82:523–531
Sineshchekov OA, Govorunova EG, Der A, Keszthelyi L, Nultsch W (1992) Photoelectric responses in phototactic flagellated algae measured in cell-suspension. J Photoch Photobio B Biol 13:119–134
Sineshchekov OA, Govorunova EG, Der A, Keszthelyi L, Nultsch W (1994) Photoinduced electric currents in carotenoid-deficient Chlamydomonas mutants reconstituted with retinal and its analogs. Biophys J 66:2073–2084
Sineshchekov OA, Jung K-H, Spudich JL (2002) Two rhodopsins mediate phototaxis to low- and high-intensity light in Chlamydomonas reinhardtii. Proc Natl Acad Sci U S A 99:8689–8694
Smyth RD, Martinek GW, Ebersold WT (1975) Linkage of six genes in Chlamydomonas reinhardtii and the construction of linkage test strains. J Bacteriol 124:1615–1617
Sodeinde OA, Kindle KL (1993) Homologous recombination in the nuclear genome of Chlamydomonas reinhardtii. Proc Natl Acad Sci U S A 90:9199–9203
Stavis RL, Hirschberg R (1973) Phototaxis in Chlamydomonas reinhardtii. J Cell Biol 59:367–377
Suzuki T et al (2003) Archaeal-type rhodopsins in Chlamydomonas: model structure and intracellular localization. Biochem Biophys Res Commun 301:711–717
Ueki N et al (2016) Eyespot-dependent determination of the phototactic sign in Chlamydomonas reinhardtii. Proc Natl Acad Sci U S A 113:5299–5304
von der Heyde EL, Hallmann A (2020) Babo1, formerly Vop1 and Cop1/2, is no eyespot photoreceptor but a basal body protein illuminating cell division in Volvox carteri. Plant J 102:276–298
Wakabayashi K, King SM (2006) Modulation of Chlamydomonas reinhardtii flagellar motility by redox poise. J Cell Biol 173:743–754
Wakabayashi K, Yagi T, Kamiya R (1997) Ca2+−dependent waveform conversion in the flagellar axoneme of Chlamydomonas mutants lacking the central-pair/radial spoke system. Cell Motil Cytoskeleton 38:22–28
Wakabayashi K, Misawa Y, Mochiji S, Kamiya R (2011) Reduction-oxidation poise regulates the sign of phototaxis in Chlamydomonas reinhardtii. Proc Natl Acad Sci U S A 108:11280–11284
Wright RL, Salisbury J, Jarvik JW (1985) A nucleus-basal body connector in Chlamydomonas reinhardtii that may function in basal body localization or segregation. J Cell Biol 101:1903–1912
Yoshimura K, Kamiya R (2001) The sensitivity of Chlamydomonas photoreceptor is optimized for the frequency of cell body rotation. Plant Cell Physiol 42:665–672
Zhang F, Wang LP, Boyden ES, Deisseroth K (2006) Channelrhodopsin-2 and optical control of excitable cells. Nat Methods 3:785–792
Acknowledgments
We would like to thank Dr. Ritsu Kamiya (Univ. Tokyo, Chuo Univ.) for critical reading of this manuscript, and Dr. Masafumi Hirono (Hosei Univ.) and Mr. Satoaki So (Tokyo Tech) for discussion. CC-5499 was made by Ms. Olga Baidukova in the Peter Hegemann’s Laboratory in Humboldt University of Berlin, and purchased from Chlamydomonas Resource Center. This work was supported by Japan Society for the Promotion of Science KAKENHI Grants 19H03242 to KW, 19K23758 to NU, by Ohsumi Frontier Science Foundation to KW, by Dynamic Alliance for Open Innovation Bridging Human, Environment and Materials to KW, and by the Assistant Staffing Program by the Gender Equality Promotion Section, Office of Public Engagement, Tokyo Institute of Technology to AI and KW.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2021 Springer Nature Singapore Pte Ltd.
About this chapter
Cite this chapter
Wakabayashi, i., Isu, A., Ueki, N. (2021). Channelrhodopsin-Dependent Photo-Behavioral Responses in the Unicellular Green Alga Chlamydomonas reinhardtii. In: Yawo, H., Kandori, H., Koizumi, A., Kageyama, R. (eds) Optogenetics. Advances in Experimental Medicine and Biology, vol 1293. Springer, Singapore. https://doi.org/10.1007/978-981-15-8763-4_2
Download citation
DOI: https://doi.org/10.1007/978-981-15-8763-4_2
Published:
Publisher Name: Springer, Singapore
Print ISBN: 978-981-15-8762-7
Online ISBN: 978-981-15-8763-4
eBook Packages: Biomedical and Life SciencesBiomedical and Life Sciences (R0)