Summary
Isolated transverse flagella ofPeridinium inconspicuum (Dinophyceae) undergo a rapid Ca2+-induced (50μM Ca2+) contraction in the absence of exogenous ATP. Longitudinal flagella from the same species do not contract under these conditions. Contraction leads to a supercoiling of the axoneme and a shortening of the paraxonemal fiber that accompanies the axoneme over most of its length. Using a polyclonal antibody generated against centrin, a 20 kDa Ca2+-modulated contractile protein of striated flagellar roots of the green flagellateTetraselmis striata, we have found that the paraxonemal fiber in transverse flagella of three taxa ofDinophyceae is immunoreactive by indirect immunofluorescence. The localization of the antigen in the paraxonemal fiber of transverse flagella was confirmed by two-colour double immunofluorescence using monoclonal mouse-anti-β-tubulin for identification of the axoneme. No structure was immunoreactive to anticentrin in the longitudinal flagella of all taxa. Electrophoretic and immunoblot analysis of isolated flagella ofP. inconspicuum show that the antigen is a 21 kDa protein, indicating that it is either centrin or a closely related protein. We conclude that centrin confers contractility to the transverse flagellum of dinoflagellates and possibly to other contractile eukaryotic flagella.
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Abbreviations
- ASP-H:
-
artificial seawater medium with Hepes-buffer
- BSA:
-
bovine serum albumine
- DTT:
-
dithiothreitol
- EGTA:
-
ethylene glycol bis(2-amino-ethylether)tetraacetic acid
- FITC:
-
fluorescein isothiocyanate
- MT:
-
buffer microtubule stabilizing buffer
- PBS:
-
phosphate buffered saline
- SDS:
-
sodium dodecyl sulfate
- TLCK:
-
Nα-p-tosyl-l-lysine chloromethyl ketone
- TRITC:
-
tetramethylrhodamine isothiocyanate
References
Afzelius BA (1969) Ultrastructure of cilia and flagella. In:Lima-de-Faria A (ed) Handbook of molecular cytology. North Holland, Amsterdam, pp 1219–1241
Amos WB (1971) Reversible mechanochemical cycle in the contraction ofVorticella. Nature 229: 127–128
—,Routledge LM, Yew FF (1975) Calcium-binding proteins in a vorticellid contractile organeile. J Cell Sci 19: 203–213
Berdach JT (1977)In situ preservation of the transverse flagellum ofPeridinium cinctum (Dinophyceae) for scanning electron microscopy. J Phycol 13: 243–251
Bessen M, Fay RB, Witman GB (1980) Calcium control of waveform in isolated axonemes ofChlamydomonas. J Cell Biol 86: 446–455
Bloodgood RA (1987) Glycoprotein dynamics in theChlamydomonas flagellar membrane. Adv Cell Biol 1: 97–130
Cachon J, Cachon M (1984) A new Ca2+-dependent function of flagellar rootlets in dinoflagellates: the releasing of a parasite from its host. Biol Cell 52: 61–76
— — (1985) Non-actin filaments and cell contraction inKofoidinium and other dinoflagellates. Cell Motil 5: 1–15
— —,Boillot A (1983) Flagellar rootlets as myonemal elements for pusule contractility in dinoflagellates. Cell Motil 3: 61–77
Cachon JM, Cachon M (1981) Movement by non-actin filament mechanisms. BioSystems 14: 313–326
Coling DE, Salisbury JL (1987) Purification and characterization of centrin, a novel calcium-modulated contractile protein. J Cell Biol 105: 1156 (Abstr)
Forward RB (1974) Phototaxis by the marine dinoflagellateGymnodinium splendens Lebour. J Protozool 21: 321–315
Gaines G, Taylor FJR (1985) Form and function of the dinoflagellate transverse flagellum. J Protozool 32: 290–296
Gibbons IR (1981) Cilia and flagella of eukaryotes. J Cell Biol 91: 107–124
—,Hiramoto Y, Mohri H, Satir P (1985) Fundamental problems of movement of cilia, eucaryotic flagella, and related systems: A seminar held under the U.S.-Japan cooperative science program. Cell Motil 5: 137–173
Gray J (1928) Ciliary movement. Cambridge University Press, London
Hand WG, Schmidt JA (1975) Phototactic orientation by the marine dinoflagellateGyrodinium dorsum Kofoid. II. Flagellar activity and overall response mechanisms. J Protozool 22: 494–498
Hoffmann-Berling H (1958) Der Mechanismus eines neuen, von der Muskelkontraktion verschiedenen Kontraktionszyklus. Biochim Biophys Acta 27: 247–255
Huang B, Mengerson A, Lee VD, Schibler M (1987) Isolation, purification, and molecular cloning of a basal body-associated calcium-binding protein fromChlamydomonas. J Cell Biol 105: 694 (Abstr)
—,Pitelka DR (1973) The contractile process in the ciliate,Stentor coeruleus. I. The role of microtubules and filaments. J Cell Biol 57: 704–728
Jahn TL, Bovee EC (1967) Motile behaviour of protozoa. In:Chen TT (ed) Research in protozoology. Pergamon Press, Oxford, pp 41–200
—,Harmon WH, Landman M (1963) Mechanisms of locomotion in flagellates. I.Ceratium. J Protozool 10: 358–363
Johnson KA (1985) Pathway of the microtubule-dynein ATPase and the structure of dynein: a comparison with actomyosin. Ann Rev Biophys Biophys Chem 14: 161–189
Laemmli UK (1970) Cleavage of structural proteins during assembly of the head of the bacteriophage T4. Nature 277: 680–685
LeBlond PH, Taylor FJB (1976) The propulsive mechanisms of the dinoflagellate transverse flagellum reconsidered. BioSystems 8: 33–39
Machemer H (1986) Electromotor coupling in cilia. Fortschr Zool 33: 205–250
Maruyama T (1981) Motion of the longitudinal flagellum inCeratium tripos (Dinoflagellida)-a retractile flagellar motion. J Protozool 28: 328–336
— (1982) Fine structure of the longitudinal flagellum inCeratium tripos, a marine dinoflagellate. J Cell Sci 58: 109–123
— (1985 a) Extraction model of the longitudinal flagellum ofCeratium tripos (Dinoflagellida): reactivation of flagellar retraction. J Cell Sci 75: 313–328
— (1985 b) Ionic control of the longitudinal flagellum inCeratium tripos (Dinoflagellida). J Protozool 32: 106–110
McFadden GI, Melkonian M (1986 a) Golgi apparatus activity and membrane flow during scale biogenesis in the green flagellateScherffelia dubia (Prasinophyceae). I.: Flagellar regeneration. Protoplasma 130: 186–198
— — (1986 b) Use of Hepes buffer for microalgal culture media and fixation for electron microscopy. Phycologia 25: 551–557
—,Schulze D, Surek B, Salisbury JL, Melkonian M (1987) Basal body reorientation mediated by a Ca2+-modulated contractile protein. J Cell Biol 105: 903–912
Melkonian M (1982) Effect of divalent cations on flagellar scales in the green flagellateTetraselmis cordifomis. Protoplasma 111: 221–233
—,Schulze D, McFadden GI, Robenek H (1988) A polyclonal antibody (anti-centrin) distinguishes between two types of fibrous flagellar roots in green algae. Protoplasma 144: 56–61
Metzner P (1929) Bewegungsstudien an Peridineen. Z Bot 22: 225–265
Moestrup Ø (1982) Flagellar structure in algae: a review with new observations particularly on the Chrysophyceae, Phaeophyceae (Fucophyceae), Euglenophyceae andReckertia. Phycologia 21: 427–528
Naitoh Y, Sugino K (1984) Ciliary movement and its control inParamecium. J Protozool 31: 31–40
Peters H (1929) Über Orts- und Geisselbewegung bei marinen Dinoflagellaten. Arch Protistenkd 67: 291–321
Rees AJJ, Leedale GF (1980) The dinoflagellate transverse flagellum: three-dimensional reconstructions from serial sections. J Phycol 16: 73–80
Salisbury JL (1983) Contractile flagellar roots: the role of calcium. J Submicrosc Cytol 15: 105–110
—,Baron A, Surek B, Melkonian M (1984) Striated flagellar roots: isolation and partial characterization of a calcium modulated contractile organelle. J Cell Biol 99: 962–970
—,Baron AT, Coling DE, Martindale VE, Sanders MA (1986) Calcium-modulated contractile proteins associated with the Eucaryotic centrosome. Cell Motil 6: 193–197
—,Floyd GL (1978) Calcium-induced contraction of the rhizoplast of a quadriflagellate green alga. Science 202: 975–977
—,Sanders MA, Harpst L (1987) Flagellar root contraction and nuclear movement during flagellar regeneration inChlamydomonas reinhardtii. J Cell Biol 105: 1799–1805
Satir P (1985) Switching mechanisms in the control of ciliary motility. In:Satir BH (ed) Modern cell biology, vol 4. Alan R Liss, New York, pp 1–46
Schulze D, Robenek H, McFadden GI, Melkonian M (1987) Immunolocalization of a Ca2+-modulated contractile protein in the flagellar apparatus of green algae: the nucleus-basal body connector. Eur J Cell Biol 45: 51–61
Schütt F (1895) Der Peridineen der Plankton-Expedition. I. Studien über die Zellen der Peridineen. Ergebn D Plankton-Exped (Kiel und Leipzig) 4: 1–170
Sleigh MA (1974) Patterns of movement of cilia and flagella. In:Sleigh MA (ed) Cilia and Flagella. Academic Press, New York, pp 79–92
Starr RC, Zeikus JA (1987) UTEX-the culture collection of algae at the University of Texas. J Phycol [Suppl] 23: 1–47
Surek B, Latzko E (1984) Visualization of antigenic proteins blotted onto nitrocellulose using the immuno-gold-staining (IGS)-method. Biochem Biophys Res Commun 121: 284–289
Taylor FJR (1975) Non-helical transverse flagella in dinoflagellates. Phycologia 14: 45–47
— (1988) Dinoflagellates. In:Margulis L, Corliss JO, Melkonian M, Chapman DL (eds) Handbook of protoctista. Jones Bartlett, Boston (in press)
Towbin H, Staehlin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76: 4350–4354
Wright BL, Salisbury J, Jarvik JW (1985) A nucleus-basal body connector inChlamydomonas reinhardtii that may function in basal body localization or segregation. J Cell Biol 101: 1903–1912
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Höhfeld, I., Otten, J. & Melkonian, M. Contractile eukaryotic flagella: Centrin is involved. Protoplasma 147, 16–24 (1988). https://doi.org/10.1007/BF01403874
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DOI: https://doi.org/10.1007/BF01403874