Skip to main content
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1976 Jan 1;68(1):48–69. doi: 10.1083/jcb.68.1.48

Mating in Chlamydomonas: a system for the study of specific cell adhesion. I. Ultrastructural and electrophoretic analyses of flagellar surface components involved in adhesion

PMCID: PMC2109620  PMID: 1245545

Abstract

To determine the ultrastructural and biochemical bases for flagellar adhesiveness in the mating reaction in Chlamydomonas, gametic and vegetative flagella and flagellar membranes were studied by use of electron microscope and electrophoretic procedures. Negative staining with uranyl acetate revealed no differences in gametic and vegetative flagellar surfaces; both had flagellar membranes, flagellar sheaths, and similar numbers and distributions of mastigonemes. Freezecleave procedures suggested that there may be a greater density of intramembranous particles on the B faces of gametic flagellar membranes than on the B faces of vegetative flagellar membranes. Gamone, the adhesive material that gametes release into their medium, was demonstrated, on the basis of ultrastructural and biochemical analyses, to be composed of flagellar surface components, i.e., membrane vesicles and mastigonemes. Comparison of vegetative (nonadhesive) and gametic (adhesive) "gamones" by use of SDS polyacrylamide gel electrophoresis showed both preparations to be composed of membrane, mastigoneme, and some microtubule proteins, as well as several unidentified protein and carbohydrate-staining components. However, there was an additional protein of approximately 70,000 mol wt in gametic gamone which was not present in vegetative gamone. When gametic gamone was separated into a membrane and a mastigoneme fraction on CSCl gradients, only the membrane fraction had isoagglutinating activity; the mastigoneme fraction was inactive, suggesting that mastigonemes are not involved in adhesion.

Full Text

The Full Text of this article is available as a PDF (7.3 MB).

Selected References

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

  1. Balsamo J., Lilien J. Embryonic cell aggregation: kinetics and specificity of binding of enhancing factors. Proc Natl Acad Sci U S A. 1974 Mar;71(3):727–731. doi: 10.1073/pnas.71.3.727. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Beug H., Katz F. E., Stein A., Gerisch G. Quantitation of membrane sites in aggregating Dictyostelium cells by use of tritiated univalent antibody. Proc Natl Acad Sci U S A. 1973 Nov;70(11):3150–3154. doi: 10.1073/pnas.70.11.3150. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cauldwell C. B., Henkart P., Humphreys T. Physical properties of sponge aggregation factor. A unique proteoglycan complex. Biochemistry. 1973 Jul 31;12(16):3051–3055. doi: 10.1021/bi00740a017. [DOI] [PubMed] [Google Scholar]
  4. Chiang K. S., Kates J. R., Jones R. F., Sueoka N. On the formation of a homogeneous zygotic population in Chlamydomonas reinhardtii. Dev Biol. 1970 Aug;22(4):655–669. doi: 10.1016/0012-1606(70)90174-0. [DOI] [PubMed] [Google Scholar]
  5. Claes H. Autolyse der Zellwand bei den Gemeten von Chlamydomonas reinhardii. Arch Mikrobiol. 1971;78(2):180–188. [PubMed] [Google Scholar]
  6. Crandall M. A., Brock T. D. Molecular aspects of specific cell contact. Science. 1968 Aug 2;161(3840):473–475. doi: 10.1126/science.161.3840.473. [DOI] [PubMed] [Google Scholar]
  7. Cunningham W. P., Staehelin L. A., Rubin R. W., Wilkins R., Bonneville M. Effects of phosphotungstate negative staining on the morphology of the isolated Golgi apparatus. J Cell Biol. 1974 Aug;62(2):491–504. doi: 10.1083/jcb.62.2.491. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Davies D. R. Electrophoretic analyses of wall glycoproteins in normal and mutant cells. Exp Cell Res. 1972 Aug;73(2):512–516. doi: 10.1016/0014-4827(72)90082-1. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Fairbanks G., Steck T. L., Wallach D. F. Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry. 1971 Jun 22;10(13):2606–2617. doi: 10.1021/bi00789a030. [DOI] [PubMed] [Google Scholar]
  11. Friedmann I., Colwin A. L., Colwin L. H. Fine-structural aspects of fertilization in Chlamydomonas reinhardi. J Cell Sci. 1968 Mar;3(1):115–128. doi: 10.1242/jcs.3.1.115. [DOI] [PubMed] [Google Scholar]
  12. Gregg J. H., Nesom M. G. Response of Dictyostelium plasma membranes to adenosine 3':5'-cyclic monophosphate. Proc Natl Acad Sci U S A. 1973 Jun;70(6):1630–1633. doi: 10.1073/pnas.70.6.1630. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Johnson U. G., Porter K. R. Fine structure of cell division in Chlamydomonas reinhardi. Basal bodies and microtubules. J Cell Biol. 1968 Aug;38(2):403–425. doi: 10.1083/jcb.38.2.403. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. KATES J. R., JONES R. F. THE CONTROL OF GAMETIC DIFFERENTIATION IN LIQUID CULTURES OF CHLAMYDOMONAS. J Cell Physiol. 1964 Apr;63:157–164. doi: 10.1002/jcp.1030630204. [DOI] [PubMed] [Google Scholar]
  15. LEVINE R. P., EBERSOLD W. T. The genetics and cytology of Chlamydomonas. Annu Rev Microbiol. 1960;14:197–216. doi: 10.1146/annurev.mi.14.100160.001213. [DOI] [PubMed] [Google Scholar]
  16. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  17. Lillie F. R. THE MECHANISM OF FERTILIZATION. Science. 1913 Oct 10;38(980):524–528. doi: 10.1126/science.38.980.524. [DOI] [PubMed] [Google Scholar]
  18. McClay D. R., Moscona A. A. Purification of the specific cell-aggregating factor from embryonic neural retina cells. Exp Cell Res. 1974 Aug;87(2):438–443. doi: 10.1016/0014-4827(74)90514-x. [DOI] [PubMed] [Google Scholar]
  19. Moscona A. A. STUDIES ON CELL AGGREGATION: DEMONSTRATION OF MATERIALS WITH SELECTIVE CELL-BINDING ACTIVITY. Proc Natl Acad Sci U S A. 1963 May;49(5):742–747. doi: 10.1073/pnas.49.5.742. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Moscona A. THE DEVELOPMENT IN VITRO OF CHIMERIC AGGREGATES OF DISSOCIATED EMBRYONIC CHICK AND MOUSE CELLS. Proc Natl Acad Sci U S A. 1957 Jan 15;43(1):184–194. doi: 10.1073/pnas.43.1.184. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]
  22. Roberts K., Gurney-Smith M., Hills G. J. Structure, composition and morphogenesis of the cell wall of Chlamydomonas reinhardi. I. Ultrastructure and preliminary chemical analysis. J Ultrastruct Res. 1972 Sep;40(5):599–613. doi: 10.1016/s0022-5320(72)80046-7. [DOI] [PubMed] [Google Scholar]
  23. Rosen S. D., Kafka J. A., Simpson D. L., Barondes S. H. Developmentally regulated, carbohydrate-binding protein in Dictyostelium discoideum. Proc Natl Acad Sci U S A. 1973 Sep;70(9):2554–2557. doi: 10.1073/pnas.70.9.2554. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Roth S. A., Weston J. A. The measurement of intercellular adhesion. Proc Natl Acad Sci U S A. 1967 Sep;58(3):974–980. doi: 10.1073/pnas.58.3.974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. SAGER R., GRANICK S. Nutritional control of sexuality in Chlamydomonas reinhardi. J Gen Physiol. 1954 Jul 20;37(6):729–742. doi: 10.1085/jgp.37.6.729. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. SAGER R. Genetic systems in Chlamydomonas. Science. 1960 Nov 18;132(3438):1459–1465. doi: 10.1126/science.132.3438.1459. [DOI] [PubMed] [Google Scholar]
  27. SAGER R., PALADE G. E. Structure and development of the chloroplast in Chlamydomonas. I. The normal green cell. J Biophys Biochem Cytol. 1957 May 25;3(3):463–488. doi: 10.1083/jcb.3.3.463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. STEINBERG M. S. Reconstruction of tissues by dissociated cells. Some morphogenetic tissue movements and the sorting out of embryonic cells may have a common explanation. Science. 1963 Aug 2;141(3579):401–408. doi: 10.1126/science.141.3579.401. [DOI] [PubMed] [Google Scholar]
  29. Sattler C. A., Staehelin L. A. Ciliary membrane differentiations in Tetrahymena pyriformis. Tetrahymena has four types of cilia. J Cell Biol. 1974 Aug;62(2):473–490. doi: 10.1083/jcb.62.2.473. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Schacterle G. R., Pollack R. L. A simplified method for the quantitative assay of small amounts of protein in biologic material. Anal Biochem. 1973 Feb;51(2):654–655. doi: 10.1016/0003-2697(73)90523-x. [DOI] [PubMed] [Google Scholar]
  31. Schmeisser E. T., Baumgartel D. M., Howell S. H. Gametic differentiation in Chlamydomonas reinhardi: cell cycle dependency and rates in attainment of mating competency. Dev Biol. 1973 Mar;31(1):31–37. doi: 10.1016/0012-1606(73)90318-7. [DOI] [PubMed] [Google Scholar]
  32. Shimada Y., Moscona A. A., Fischman D. A. Scanning electron microscopy of cell aggregation: cardiac and mixed retina-cardiac cell suspensions. Dev Biol. 1974 Feb;36(2):428–446. doi: 10.1016/0012-1606(74)90063-3. [DOI] [PubMed] [Google Scholar]
  33. Smith G. M., Regnery D. C. Inheritance of Sexuality in Chlamydomonas Reinhardi. Proc Natl Acad Sci U S A. 1950 Apr;36(4):246–248. doi: 10.1073/pnas.36.4.246. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Snell W. J., Dentler W. L., Haimo L. T., Binder L. I., Rosenbaum J. L. Assembly of chick brain tubulin onto isolated basal bodies of Chlamydomonas reinhardi. Science. 1974 Jul 26;185(4148):357–360. doi: 10.1126/science.185.4148.357. [DOI] [PubMed] [Google Scholar]
  35. Snell W. J. Mating in Chlamydomonas: a system for the study of specific cell adhesion. II. A radioactive flagella-binding assay for quantitation of adhesion. J Cell Biol. 1976 Jan;68(1):70–79. doi: 10.1083/jcb.68.1.70. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Steinberg M. S. Does differential adhesion govern self-assembly processes in histogenesis? Equilibrium configurations and the emergence of a hierarchy among populations of embryonic cells. J Exp Zool. 1970 Apr;173(4):395–433. doi: 10.1002/jez.1401730406. [DOI] [PubMed] [Google Scholar]
  37. Sueoka N., Chiang K. S., Kates J. R. Deoxyribonucleic acid replication in meiosis of Chlamydomonas reinhardi. I. Isotopic transfer experiments with a strain producing eight zoospores. J Mol Biol. 1967 Apr 14;25(1):47–66. doi: 10.1016/0022-2836(67)90278-1. [DOI] [PubMed] [Google Scholar]
  38. TAYLOR N. W. SPECIFIC, SOLUBLE FACTOR INVOLVED IN SEXUAL AGGLUTINATION OF THE YEAST HANSENULA WINGEI. J Bacteriol. 1964 Apr;87:863–866. doi: 10.1128/jb.87.4.863-866.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Wiese L. Nature of sex specific glycoprotein agglutinins in Chlamydomonas. Ann N Y Acad Sci. 1974;234(0):383–395. doi: 10.1111/j.1749-6632.1974.tb53050.x. [DOI] [PubMed] [Google Scholar]
  40. Witman G. B., Carlson K., Berliner J., Rosenbaum J. L. Chlamydomonas flagella. I. Isolation and electrophoretic analysis of microtubules, matrix, membranes, and mastigonemes. J Cell Biol. 1972 Sep;54(3):507–539. doi: 10.1083/jcb.54.3.507. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Yen P. H., Ballou C. E. Composition of a specific intercellular agglutination factor. J Biol Chem. 1973 Dec 10;248(23):8316–8318. [PubMed] [Google Scholar]

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

RESOURCES