Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1980 Aug 1;86(2):529–536. doi: 10.1083/jcb.86.2.529

A collagen-binding protein on the surface of ejaculated rabbit spermatozoa

PMCID: PMC2111506  PMID: 6995466

Abstract

A rabbit antibody to mouse 3T3 cell fibronectin was used in conjunction with a fluorescein-tagged second antibody to detect fibronectin-like activity on the surface of rabbit spermatozoa. Only ejaculated sperm displayed an intense and highly localized fluorescence over the acrosomal region. Cauda epididymal sperm of the rabbit as well as several other species did not exhibit any reaction. The fluorescent activity could be eliminated by trypsin treatment but was re- established by incubation in cell-free seminal fluid. Sperm recovered from females 10-12 h after mating showed a reduction or absence of antifibronectin fluorescence, suggesting that this component's loss could be a factor in sperm capacitation. Because fibronectins show strong binding to collagen, mixtures of ejaculated sperm and collagen were examined in the light and electron microscope. Living sperm appear to have a strong affinity for collagen and quickly adhere to the filaments by their heads, while continuing vigorous flagellations. Surface labeling of sperm with the galactose-oxidase-NaB[3H]4 technique, extraction with urea-detergent mixtures and affinity chromatography of extracts on gelatin-Sepharose revealed a single radioactive band of mot wt approximately 40,000 after SDS polyacrylamide gel electrophoresis and fluorography.

Full Text

The Full Text of this article is available as a PDF (893.9 KB).

Selected References

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

  1. Balian G., Click E. M., Crouch E., Davidson J. M., Bornstein P. Isolation of a collagen-binding fragment from fibronectin and cold-insoluble globulin. J Biol Chem. 1979 Mar 10;254(5):1429–1432. [PubMed] [Google Scholar]
  2. Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
  3. Chen L. B., Burridge K., Murray A., Walsh M. L., Copple C. D., Bushnell A., McDougall J. K., Gallimore P. H. Modulation of cell surface glycocalyx: studies on large, external, transformation-sensitive protein. Ann N Y Acad Sci. 1978 Jun 20;312:366–381. doi: 10.1111/j.1749-6632.1978.tb16814.x. [DOI] [PubMed] [Google Scholar]
  4. Chvapil M. An intravaginal contraceptive diaphragm made of collagen sponge: new old principle. Fertil Steril. 1976 Dec;27(12):1387–1397. [PubMed] [Google Scholar]
  5. Chvapil M., Chvapil T., Jacobs S., Owen J. A., Horton H. L., Heine M. W. Laboratory and clinical testing of an intravaginal contraceptive collagen sponge. Contraception. 1977 Jun;15(6):693–703. doi: 10.1016/s0010-7824(77)80006-1. [DOI] [PubMed] [Google Scholar]
  6. Eng L. A., Oliphant G. Rabbit sperm reversible decapacitation by membrane stabilization with a highly purified glycoprotein from seminal plasma. Biol Reprod. 1978 Dec;19(5):1083–1094. doi: 10.1095/biolreprod19.5.1083. [DOI] [PubMed] [Google Scholar]
  7. Engvall E., Ruoslahti E. Binding of soluble form of fibroblast surface protein, fibronectin, to collagen. Int J Cancer. 1977 Jul 15;20(1):1–5. doi: 10.1002/ijc.2910200102. [DOI] [PubMed] [Google Scholar]
  8. Esbenshade K. L., Clegg E. D. Electrophoretic characterization of proteins in the plasma membrane of porcine spermatozoa. J Reprod Fertil. 1976 Jul;47(2):333–337. doi: 10.1530/jrf.0.0470333. [DOI] [PubMed] [Google Scholar]
  9. Espey L. L., Rondell P. Collagenolytic activity in the rabbit and sow Graafian follicle during ovulation. Am J Physiol. 1968 Feb;214(2):326–329. doi: 10.1152/ajplegacy.1968.214.2.326. [DOI] [PubMed] [Google Scholar]
  10. Fukuda M., Hakomori S. Proteolytic and chemical fragmentation of galactoprotein a, a major transformation-sensitive glycoprotein released from hamster embryo fibroblasts. J Biol Chem. 1979 Jun 25;254(12):5442–5450. [PubMed] [Google Scholar]
  11. Gahmberg C. G., Hakomori S. I. Altered growth behavior of malignant cells associated with changes in externally labeled glycoprotein and glycolipid. Proc Natl Acad Sci U S A. 1973 Dec;70(12):3329–3333. doi: 10.1073/pnas.70.12.3329. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gahmberg C. G., Hakomori S. I. External labeling of cell surface galactose and galactosamine in glycolipid and glycoprotein of human erythrocytes. J Biol Chem. 1973 Jun 25;248(12):4311–4317. [PubMed] [Google Scholar]
  13. Hahn L. H., Yamada K. M. Identification and isolation of a collagen-binding fragment of the adhesive glycoprotein fibronectin. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1160–1163. doi: 10.1073/pnas.76.3.1160. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hayman E. G., Ruoslahti E. Distribution of fetal bovine serum fibronectin and endogenous rat cell fibronectin in extracellular matrix. J Cell Biol. 1979 Oct;83(1):255–259. doi: 10.1083/jcb.83.1.255. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hunter A. G., Nornes H. O. Characterization and isolation of a sperm-coating antigen from rabbit seminal plasma with capacity to block fertilization. J Reprod Fertil. 1969 Dec;20(3):419–427. doi: 10.1530/jrf.0.0200419. [DOI] [PubMed] [Google Scholar]
  16. Hynes R. O. Alteration of cell-surface proteins by viral transformation and by proteolysis. Proc Natl Acad Sci U S A. 1973 Nov;70(11):3170–3174. doi: 10.1073/pnas.70.11.3170. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Johnson W. L., Hunter A. G. Seminal antigens: their alteration in the genital tract of female rabbits and during partial in vitro capacitation with beta amylase and beta glucuronidase. Biol Reprod. 1972 Dec;7(3):332–340. doi: 10.1093/biolreprod/7.3.332. [DOI] [PubMed] [Google Scholar]
  18. Kaplan J. E., Saba T. M. Humoral deficiency and reticuloendothelial depression after traumatic shock. Am J Physiol. 1976 Jan;230(1):7–14. doi: 10.1152/ajplegacy.1976.230.1.7. [DOI] [PubMed] [Google Scholar]
  19. Koren E., Milković S. 'Collagenase-like peptidase in human, rat and bull spermatozoa. J Reprod Fertil. 1973 Mar;32(3):349–356. doi: 10.1530/jrf.0.0320349. [DOI] [PubMed] [Google Scholar]
  20. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  21. Linder E., Vaheri A., Ruoslahti E., Wartiovaara J. Distribution of fibroblast surface antigen in the developing chick embryo. J Exp Med. 1975 Jul 1;142(1):41–49. doi: 10.1084/jem.142.1.41. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Mayol R. F., Longenecker D. E. Separation of non-sperm components from seminal preparations and effect on the analysis of sperm proteins. Proc Soc Exp Biol Med. 1975 May;149(1):64–69. doi: 10.3181/00379727-149-38744. [DOI] [PubMed] [Google Scholar]
  23. Nicander L., Plöen L., Larsson M. Specific apocrine secretion in the anterior lobe of the prostate gland of rabbits. Cell Tissue Res. 1974;151(1):69–77. doi: 10.1007/BF00222035. [DOI] [PubMed] [Google Scholar]
  24. Oliphant G., Singhas C. A. Iodination of rabbit sperm plasma membrane: relationship of specific surface proteins to epididymal function and sperm capacitation. Biol Reprod. 1979 Nov;21(4):937–944. doi: 10.1095/biolreprod21.4.937. [DOI] [PubMed] [Google Scholar]
  25. Olson G. E., Hamilton D. W. Characterization of the surface glycoproteins of rat spermatozoa. Biol Reprod. 1978 Aug;19(1):26–35. doi: 10.1095/biolreprod19.1.26. [DOI] [PubMed] [Google Scholar]
  26. Pearlstein E. Plasma membrane glycoprotein which mediates adhesion of fibroblasts to collagen. Nature. 1976 Aug 5;262(5568):497–500. doi: 10.1038/262497a0. [DOI] [PubMed] [Google Scholar]
  27. Schwarz M. A., Koehler J. K. Alterations in lectin binding to guinea pig spermatozoa accompanying in vitro capacitation and the acrosome reaction. Biol Reprod. 1979 Dec;21(5):1295–1307. doi: 10.1095/biolreprod21.5.1295. [DOI] [PubMed] [Google Scholar]
  28. Stenman S., Vaheri A. Distribution of a major connective tissue protein, fibronectin, in normal human tissues. J Exp Med. 1978 Apr 1;147(4):1054–1064. doi: 10.1084/jem.147.4.1054. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Strickland S., Beers W. H. Studies on the role of plasminogen activator in ovulation. In vitro response of granulosa cells to gonadotropins, cyclic nucleotides, and prostaglandins. J Biol Chem. 1976 Sep 25;251(18):5694–5702. [PubMed] [Google Scholar]
  30. Vaheri A., Ruoslahti E. Disappearance of a major cell-type specific surface glycoprotein antigen (SF) after transformation of fibroblasts by Rous sarcoma virus. Int J Cancer. 1974 May 15;13(5):579–586. doi: 10.1002/ijc.2910130502. [DOI] [PubMed] [Google Scholar]
  31. Wartiovaara J., Leivo I., Virtanen I., Vaheri A., Graham C. F. Cell surface and extracellular matrix glycoprotein fibronectin: expression in embryogenesis and in teratocarcinoma differentiation. Ann N Y Acad Sci. 1978 Jun 20;312:132–141. doi: 10.1111/j.1749-6632.1978.tb16798.x. [DOI] [PubMed] [Google Scholar]
  32. Yamada K. M., Olden K. Fibronectins--adhesive glycoproteins of cell surface and blood. Nature. 1978 Sep 21;275(5677):179–184. doi: 10.1038/275179a0. [DOI] [PubMed] [Google Scholar]
  33. Yamada K. M., Weston J. A. Isolation of a major cell surface glycoprotein from fibroblasts. Proc Natl Acad Sci U S A. 1974 Sep;71(9):3492–3496. doi: 10.1073/pnas.71.9.3492. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Zetter B. R., Martin G. R., Birdwell C. R., Gospodarowicz D. Role of the high-molecular-weight glycoprotein in cellular morphology, adhesion, and differentiation. Ann N Y Acad Sci. 1978 Jun 20;312:299–316. doi: 10.1111/j.1749-6632.1978.tb16810.x. [DOI] [PubMed] [Google Scholar]

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

RESOURCES