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. 1982 Nov 1;95(2):567–573. doi: 10.1083/jcb.95.2.567

Sperm surface galactosyltransferase activities during in vitro capacitation

BD Shur, NG Hall
PMCID: PMC2112945  PMID: 6815211

Abstract

Studies using genetic and biochemical probes have suggested that mouse sperm surface galactosyltransferases may participate during fertilization by binding N- acetylglucosamine (GlcNAc) residues in the egg zona pellucida. In light of these results, we examined sperm surface galactosyltransferase activity during in vitro capacitation to determine whether changes in enzymatic activity correlated with fertilizing ability. Results show that surface galactosyltransferases on uncapacitated sperm was preferentially loaded with poly N-acetyllactosamine substrates. As a consequence of capacitation in Ca(++)-containing medium, these polylactosaminyl substrates are spontaneously released from the sperm surface, thereby exposing the sperm galactosyltransferase for binding to the zona pellucida. Sperm capacitation can be mimicked, in the absence of Ca(++), either by washing sperm in Ca(++)-free medium, or by pretreating sperm with antiserum that reacts with the galactosyltransferase substrate. In both instances, sperm galgactosylation of endogenous polylactosaminyl substrates is reduced, coincident with increased galactosylation of exogenous GlcNAc, and increased binding to the zona pellucida. Binding of capacitated sperm to the egg can be inhibited by pronase-digested high molecular weight polyactosaminyl glycoside extracted from epidymal fluids or from undifferentiated F9 embryonal carninoma cells. Thus, these glycosides function as “decapacitation factors” when added back to in vitro fertilization assays. These glycoside “decapacitation factors” inhibit sperm-egg binding by competeing for the sperm surface galactosyltransferase, since (a) they are galactosylated by sperm in the presence of UDP[(3)H]galactose, and (b) enzymatic removal of terminal GlcNAc residues reduces “decapacitation factio” competition. On the other hand “conventional” low molecular weight glycosides, isolated from either epididymal fluid or differentiated F9 cells, fail to inhibit capacitated sperm binding to the zona pellucida. These results define a molecular mechanism for one aspect of sperm capacitation, and help explain why removal of “decapacitation factos” is a necessary prerequisite for sperm binding to the zona pellucida.

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Selected References

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

  1. Acott T. S., Hoskins D. D. Bovine sperm forward motility protein. Partial purification and characterization. J Biol Chem. 1978 Oct 10;253(19):6744–6750. [PubMed] [Google Scholar]
  2. Artzt K., Dubois P., Bennett D., Condamine H., Babinet C., Jacob F. Surface antigens common to mouse cleavage embryos and primitive teratocarcinoma cells in culture. Proc Natl Acad Sci U S A. 1973 Oct;70(10):2988–2992. doi: 10.1073/pnas.70.10.2988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. BRINSTER R. L. STUDIES ON THE DEVELOPMENT OF MOUSE EMBRYOS IN VITRO. II. THE EFFECT OF ENERGY SOURCE. J Exp Zool. 1965 Feb;158:59–68. doi: 10.1002/jez.1401580106. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Gwatkin R. B., Andersen O. F. Effect of glycosidase inhibitors on the capacitation of hamster spermatozoa by cumulus cells in vitro. J Reprod Fertil. 1973 Dec;35(3):565–567. doi: 10.1530/jrf.0.0350565. [DOI] [PubMed] [Google Scholar]
  5. Kinsey W. H., Lennarz W. J. Isolation of a glycopeptide fraction from the surface of the sea urchin egg that inhibits sperm-egg binding and fertilization. J Cell Biol. 1981 Nov;91(2 Pt 1):325–331. doi: 10.1083/jcb.91.2.325. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Muramatsu T., Gachelin G., Damonneville M., Delarbre C., Jacob F. Cell surface carbohydrates of embryonal carcinoma cells: polysaccharidic side chains of F9 antigens and of receptors to two lectins, FBP and PNA. Cell. 1979 Sep;18(1):183–191. doi: 10.1016/0092-8674(79)90367-2. [DOI] [PubMed] [Google Scholar]
  7. Nakazawa K., Suzuki S. Purification of Keratan Sulfate-endogalactosidase and its action on keratan sulfates of different origin. J Biol Chem. 1975 Feb 10;250(3):912–917. [PubMed] [Google Scholar]
  8. Oliphant G., Brackett B. G. Capacitation of mouse spermatozoa in media with elevated ionic strength and reversible decapacitation with epididymal extracts. Fertil Steril. 1973 Dec;24(12):948–955. doi: 10.1016/s0015-0282(16)40094-4. [DOI] [PubMed] [Google Scholar]
  9. Robbins P. W., Krag S. S., Liu T. Effects of UDP-glucose addition on the synthesis of mannosyl lipid-linked oligosaccharides by cell-free fibroblast preparations. J Biol Chem. 1977 Mar 10;252(5):1780–1785. [PubMed] [Google Scholar]
  10. Saling P. M., Storey B. T. Mouse gamete interactions during fertilization in vitro. Chlortetracycline as a fluorescent probe for the mouse sperm acrosome reaction. J Cell Biol. 1979 Dec;83(3):544–555. doi: 10.1083/jcb.83.3.544. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Saling P. M., Storey B. T., Wolf D. P. Calcium-dependent binding of mouse epididymal spermatozoa to the zona pellucida. Dev Biol. 1978 Aug;65(2):515–525. doi: 10.1016/0012-1606(78)90046-5. [DOI] [PubMed] [Google Scholar]
  12. Shapiro B. M., Eddy E. M. When sperm meets egg: biochemical mechanisms of gamete interaction. Int Rev Cytol. 1980;66:257–302. doi: 10.1016/s0074-7696(08)61976-2. [DOI] [PubMed] [Google Scholar]
  13. Shur B. D., Bennett D. A specific defect in galactosyltransferase regulation on sperm bearing mutant alleles of the T/t locus. Dev Biol. 1979 Aug;71(2):243–259. doi: 10.1016/0012-1606(79)90167-2. [DOI] [PubMed] [Google Scholar]
  14. Shur B. D. Cell surface glycosyltransferase activities during normal and mutant (T/T) mesenchyme migration. Dev Biol. 1982 May;91(1):149–162. doi: 10.1016/0012-1606(82)90018-5. [DOI] [PubMed] [Google Scholar]
  15. Shur B. D. Evidence that galactosyltransferase is a surface receptor for poly(N)-acetyllactosamine glycoconjugates on embryonal carcinoma cells. J Biol Chem. 1982 Jun 25;257(12):6871–6878. [PubMed] [Google Scholar]
  16. Shur B. D. Galactosyltransferase activities on mouse sperm bearing multiple tlethal and tviable haplotypes of the T/t-complex. Genet Res. 1981 Dec;38(3):225–236. doi: 10.1017/s0016672300020577. [DOI] [PubMed] [Google Scholar]
  17. Shur B. D., Hall N. G. A role for mouse sperm surface galactosyltransferase in sperm binding to the egg zona pellucida. J Cell Biol. 1982 Nov;95(2 Pt 1):574–579. doi: 10.1083/jcb.95.2.574. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Shur B. D., Roth S. Cell surface glycosyltransferases. Biochim Biophys Acta. 1975 Dec 29;415(4):473–512. doi: 10.1016/0304-4157(75)90007-6. [DOI] [PubMed] [Google Scholar]
  19. Vacquier V. D., Moy G. W. Isolation of bindin: the protein responsible for adhesion of sperm to sea urchin eggs. Proc Natl Acad Sci U S A. 1977 Jun;74(6):2456–2460. doi: 10.1073/pnas.74.6.2456. [DOI] [PMC free article] [PubMed] [Google Scholar]

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