Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1983 Jun 1;96(6):1610–1621. doi: 10.1083/jcb.96.6.1610

Motile detergent-extracted cells of Tetrahymena and Chlamydomonas

PMCID: PMC2112455  PMID: 6602134

Abstract

Tetrahymena and Chlamydomonas cells treated with high (0.25-0.5%) concentrations of the detergent Nonidet P-40 in appropriate buffers retain the shape of the intact cells but are devoid of any ciliary activity unless supplied with MgATP. ATP causes them to swim actively, with beat parameters and swimming patterns indistinguishable from those of intact cells. Both types of detergent-extracted cells are completely devoid of ciliary membranes. The Tetrahymena preparations also lack all cellular membranes, whereas cellular membranes remain intact in the Chlamydomonas preparations. Experiments demonstrating the effects of ATP, ADP, vanadate, erythro-9-[3-2-(hydroxynonyl)]-adenine, and Ca++ are described to illustrate the use of these detergent-extracted cells in research on ciliary motility.

Full Text

The Full Text of this article is available as a PDF (7.0 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Allen C., Borisy G. G. Structural polarity and directional growth of microtubules of Chlamydomonas flagella. J Mol Biol. 1974 Dec 5;90(2):381–402. doi: 10.1016/0022-2836(74)90381-7. [DOI] [PubMed] [Google Scholar]
  2. Allen R. D. Fine structure, reconstruction and possible functions of components of the cortex of Tetrahymena pyriformis. J Protozool. 1967 Nov;14(4):553–565. doi: 10.1111/j.1550-7408.1967.tb02042.x. [DOI] [PubMed] [Google Scholar]
  3. Asai D. J., Brokaw C. J. Effects of antibodies against tubulin on the movement of reactivated sea urchin sperm flagella. J Cell Biol. 1980 Oct;87(1):114–123. doi: 10.1083/jcb.87.1.114. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bessen M., Fay R. B., Witman G. B. Calcium control of waveform in isolated flagellar axonemes of Chlamydomonas. J Cell Biol. 1980 Aug;86(2):446–455. doi: 10.1083/jcb.86.2.446. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Blake J. R., Sleigh M. A. Mechanics of ciliary locomotion. Biol Rev Camb Philos Soc. 1974 Feb;49(1):85–125. doi: 10.1111/j.1469-185x.1974.tb01299.x. [DOI] [PubMed] [Google Scholar]
  6. Bouchard P., Penningroth S. M., Cheung A., Gagnon C., Bardin C. W. erythro-9-[3-(2-Hydroxynonyl)]adenine is an inhibitor of sperm motility that blocks dynein ATPase and protein carboxylmethylase activities. Proc Natl Acad Sci U S A. 1981 Feb;78(2):1033–1036. doi: 10.1073/pnas.78.2.1033. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Brokaw C. J., Gibbons I. R. Localized activation of bending in proximal, medial and distal regions of sea-urchin sperm flagella. J Cell Sci. 1973 Jul;13(1):1–10. doi: 10.1242/jcs.13.1.1. [DOI] [PubMed] [Google Scholar]
  8. Brokaw C. J., Luck D. J., Huang B. Analysis of the movement of Chlamydomonas flagella:" the function of the radial-spoke system is revealed by comparison of wild-type and mutant flagella. J Cell Biol. 1982 Mar;92(3):722–732. doi: 10.1083/jcb.92.3.722. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Bruns P. J., Brussard T. E. Nullisomic tetrahymena: eliminating germinal chromosomes. Science. 1981 Jul 31;213(4507):549–551. doi: 10.1126/science.213.4507.549. [DOI] [PubMed] [Google Scholar]
  10. Dentler W. L., LeCluyse E. L. Microtubule capping structures at the tips of tracheal cilia: evidence for their firm attachment during ciliary bend formation and the restriction of microtubule sliding. Cell Motil. 1982;2(6):549–572. doi: 10.1002/cm.970020605. [DOI] [PubMed] [Google Scholar]
  11. EBERSOLD W. T., LEVINE R. P., LEVINE E. E., OLMSTED M. A. Linkage maps in Chlamydomonas reinhardi. Genetics. 1962 May;47:531–543. doi: 10.1093/genetics/47.5.531. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Eckert R., Murakami A. Calcium dependence of ciliary activity in the oviduct of the salamander Necturus. J Physiol. 1972 Nov;226(3):699–711. doi: 10.1113/jphysiol.1972.sp010004. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gibbons B. H., Gibbons I. R. Calcium-induced quiescence in reactivated sea urchin sperm. J Cell Biol. 1980 Jan;84(1):13–27. doi: 10.1083/jcb.84.1.13. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gibbons B. H., Gibbons I. R. Flagellar movement and adenosine triphosphatase activity in sea urchin sperm extracted with triton X-100. J Cell Biol. 1972 Jul;54(1):75–97. doi: 10.1083/jcb.54.1.75. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gibbons B. H., Gibbons I. R. Properties of flagellar "rigor waves" formed by abrupt removal of adenosine triphosphate from actively swimming sea urchin sperm. J Cell Biol. 1974 Dec;63(3):970–985. doi: 10.1083/jcb.63.3.970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Gibbons B. H., Gibbons I. R. The effect of partial extraction of dynein arms on the movement of reactivated sea-urchin sperm. J Cell Sci. 1973 Sep;13(2):337–357. doi: 10.1242/jcs.13.2.337. [DOI] [PubMed] [Google Scholar]
  17. Gibbons I. R. Cilia and flagella of eukaryotes. J Cell Biol. 1981 Dec;91(3 Pt 2):107s–124s. doi: 10.1083/jcb.91.3.107s. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Gibbons I. R., Cosson M. P., Evans J. A., Gibbons B. H., Houck B., Martinson K. H., Sale W. S., Tang W. J. Potent inhibition of dynein adenosinetriphosphatase and of the motility of cilia and sperm flagella by vanadate. Proc Natl Acad Sci U S A. 1978 May;75(5):2220–2224. doi: 10.1073/pnas.75.5.2220. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Goodenough U. W., Heuser J. E. Substructure of the outer dynein arm. J Cell Biol. 1982 Dec;95(3):798–815. doi: 10.1083/jcb.95.3.798. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Gorman D. S., Levine R. P. Cytochrome f and plastocyanin: their sequence in the photosynthetic electron transport chain of Chlamydomonas reinhardi. Proc Natl Acad Sci U S A. 1965 Dec;54(6):1665–1669. doi: 10.1073/pnas.54.6.1665. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Gorovsky M. A., Yao M. C., Keevert J. B., Pleger G. L. Isolation of micro- and macronuclei of Tetrahymena pyriformis. Methods Cell Biol. 1975;9(0):311–327. doi: 10.1016/s0091-679x(08)60080-1. [DOI] [PubMed] [Google Scholar]
  22. Heuser J. Three-dimensional visualization of coated vesicle formation in fibroblasts. J Cell Biol. 1980 Mar;84(3):560–583. doi: 10.1083/jcb.84.3.560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Hoffman J. L., Goodenough U. W. Experimental dissection of flagellar surface motility in Chlamydomonas. J Cell Biol. 1980 Aug;86(2):656–665. doi: 10.1083/jcb.86.2.656. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Huang B., Piperno G., Luck D. J. Paralyzed flagella mutants of Chlamydomonas reinhardtii. Defective for axonemal doublet microtubule arms. J Biol Chem. 1979 Apr 25;254(8):3091–3099. [PubMed] [Google Scholar]
  25. Huang B., Ramanis Z., Luck D. J. Suppressor mutations in Chlamydomonas reveal a regulatory mechanism for Flagellar function. Cell. 1982 Jan;28(1):115–124. doi: 10.1016/0092-8674(82)90381-6. [DOI] [PubMed] [Google Scholar]
  26. Hyams J. S., Borisy G. G. 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. 1978 Oct;33:235–253. doi: 10.1242/jcs.33.1.235. [DOI] [PubMed] [Google Scholar]
  27. Johnson K. A., Porter M. E. Transient state kinetic analysis of the dynein ATPase. Prog Clin Biol Res. 1982;80:101–106. doi: 10.1002/cm.970020720. [DOI] [PubMed] [Google Scholar]
  28. Kobayashi T., Martensen T., Nath J., Flavin M. Inhibition of dynein ATPase by vanadate, and its possible use as a probe for the role of dynein in cytoplasmic motility. Biochem Biophys Res Commun. 1978 Apr 28;81(4):1313–1318. doi: 10.1016/0006-291x(78)91279-2. [DOI] [PubMed] [Google Scholar]
  29. Martin N. C., Goodenough U. W. Gametic differentiation in Chlamydomonas reinhardtii. I. Production of gametes and their fine structure. J Cell Biol. 1975 Dec;67(3):587–605. doi: 10.1083/jcb.67.3.587. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Mesland D. A. Mating in Chlamydomonas eugametos. A scanning electron microscopical study. Arch Microbiol. 1976 Aug;109(1-2):31–35. doi: 10.1007/BF00425109. [DOI] [PubMed] [Google Scholar]
  31. Mitchell D. R., Warner F. D. Interactions of dynein arms with b subfibers of Tetrahymena cilia: quantitation of the effects of magnesium and adenosine triphosphate. J Cell Biol. 1980 Oct;87(1):84–97. doi: 10.1083/jcb.87.1.84. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Naito Y., Kaneko H. Control of ciliary activities by adenosinetriphosphate and divalent cations in triton-extracted models of Paramecium caudatum. J Exp Biol. 1973 Jun;58(3):657–676. doi: 10.1242/jeb.58.3.657. [DOI] [PubMed] [Google Scholar]
  33. Naito Y., Kaneko H. Reactivated triton-extracted models o paramecium: modification of ciliary movement by calcium ions. Science. 1972 May 5;176(4034):523–524. doi: 10.1126/science.176.4034.523. [DOI] [PubMed] [Google Scholar]
  34. Peck R. K. The ultrastructure of the somatic cortex of Pseudomicrothorax dubius: structure and function of the epiplasm in ciliated protozoa. J Cell Sci. 1977 Jun;25:367–385. doi: 10.1242/jcs.25.1.367. [DOI] [PubMed] [Google Scholar]
  35. Penningroth S. M., Cheung A., Bouchard P., Gagnon C., Bardin C. W. Dynein ATPase is inhibited selectively in vitro by erythro-9-[3-2-(hydroxynonyl)]adenine. Biochem Biophys Res Commun. 1982 Jan 15;104(1):234–240. doi: 10.1016/0006-291x(82)91964-7. [DOI] [PubMed] [Google Scholar]
  36. Piperno G., Luck D. J. Axonemal adenosine triphosphatases from flagella of Chlamydomonas reinhardtii. Purification of two dyneins. J Biol Chem. 1979 Apr 25;254(8):3084–3090. [PubMed] [Google Scholar]
  37. Piperno G., Luck D. J. Inner arm dyneins from flagella of Chlamydomonas reinhardtii. Cell. 1981 Dec;27(2 Pt 1):331–340. doi: 10.1016/0092-8674(81)90416-5. [DOI] [PubMed] [Google Scholar]
  38. Raff E. C., Blums J. J. A possible role for adenylate kinase in cilia: concentration profiles in a geometrically constrained dual enzyme system. J Theor Biol. 1968 Jan;18(1):53–71. doi: 10.1016/0022-5193(68)90170-7. [DOI] [PubMed] [Google Scholar]
  39. Ringo D. L. Flagellar motion and fine structure of the flagellar apparatus in Chlamydomonas. J Cell Biol. 1967 Jun;33(3):543–571. doi: 10.1083/jcb.33.3.543. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Rosenbaum J. L., Moulder J. E., Ringo D. L. Flagellar elongation and shortening in Chlamydomonas. The use of cycloheximide and colchicine to study the synthesis and assembly of flagellar proteins. J Cell Biol. 1969 May;41(2):600–619. doi: 10.1083/jcb.41.2.600. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Sale W. S., Gibbons I. R. Study of the mechanism of vanadate inhibition of the dynein cross-bridge cycle in sea urchin sperm flagella. J Cell Biol. 1979 Jul;82(1):291–298. doi: 10.1083/jcb.82.1.291. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Schmidt J. A., Eckert R. Calcium couples flagellar reversal to photostimulation in Chlamydomonas reinhardtii. Nature. 1976 Aug 19;262(5570):713–715. doi: 10.1038/262713a0. [DOI] [PubMed] [Google Scholar]
  43. Takahashi M., Tonomura Y. Binding of 30s dynein with the B-tubule of the outer doublet of axonemes from Tetrahymena pyriformis and adenosine triphosphate-induced dissociation of the complex. J Biochem. 1978 Dec;84(6):1339–1355. doi: 10.1093/oxfordjournals.jbchem.a132256. [DOI] [PubMed] [Google Scholar]
  44. Tokuyasu K., Scherbaum O. H. Ultrastructure of mucocysts and pellicle of Tetrahymena pyriformis. J Cell Biol. 1965 Oct;27(1):67–81. doi: 10.1083/jcb.27.1.67. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Torres L. D., Renaud F. L., Portocarrero C. Studies on reactivated cilia. II. Reactivation of ciliated cortices from the oviduct of Anolis cristatellus. Exp Cell Res. 1977 Sep;108(2):311–320. doi: 10.1016/s0014-4827(77)80038-4. [DOI] [PubMed] [Google Scholar]
  46. Walter M. F., Satir P. Calcium control of ciliary arrest in mussel gill cells. J Cell Biol. 1978 Oct;79(1):110–120. doi: 10.1083/jcb.79.1.110. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Warner F. D., Mitchell D. R. Structural conformation of ciliary dynein arms and the generation of sliding forces in Tetrahymena cilia. J Cell Biol. 1978 Feb;76(2):261–277. doi: 10.1083/jcb.76.2.261. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Williams N. E., Vaudaux P. E., Skriver L. Cytoskeletal proteins of the cell surface in Tetrahymena I. Identification and localization of major proteins. Exp Cell Res. 1979 Oct 15;123(2):311–320. doi: 10.1016/0014-4827(79)90473-7. [DOI] [PubMed] [Google Scholar]
  49. Witman G. B., Plummer J., Sander G. Chlamydomonas flagellar mutants lacking radial spokes and central tubules. Structure, composition, and function of specific axonemal components. J Cell Biol. 1978 Mar;76(3):729–747. doi: 10.1083/jcb.76.3.729. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Wolfe J. Structural analysis of basal bodies of the isolated oral apparatus of Tetrahymena pyriformis. J Cell Sci. 1970 May;6(3):679–700. doi: 10.1242/jcs.6.3.679. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES