Summary
A fucose binding, protein was detected in boar spermatozoa by means of a specifically developed modified enzyme-linked-lectin-assay using glycosylated peroxidase derivatives. The distribution of the fucose binding protein was assessed by means of fluorescence microscopy with fluoresceinyl-glycosylated peroxidase. Fucose binding was particularly prominent at the apical region of the sperm head. In order to gain more insight into the precise localization of the carbohydrate binding, protein electron microscopical studies were performed using fucosyl peroxidase coupled to colloidal gold. In ultrathin sections as well as in specimens, prepared in toto for TEM an intensive binding of fucosylperoxidase-colloidal gold was predominantly found at the apical part of the acrosome appearing as a crescent-like area. In some cases this binding pattern was replaced by a triangle-like intensive labelling at the equatorial segment as revealed clearly by specimens prepared in toto. By SDS-PAGE of the SDS-extractable sperm-proteins, followed by transblotting to nitrocellulose and visualization with the fucosylperoxidase by enzymatic amplification with 4-chloro-1-naphthol mainly one protein with the reduced molecular weight of approximately 53 kdal and some small proteins with apparent molecular weights less than 20 kdal was found to be responsible for the fucose-binding ability of porcine spermatozoa.
Similar content being viewed by others
References
Ahuja KK (1982) Fertilization studies in the hamster. The role of cell-surface carbohydrates. Exp Cell Res 140:353–362
Frazier W, Glaser L (1979) Surface and cell recognition. Annu Rev Biochem 48:491–523
Friess AE, Sinowatz F (1984) Con A-and WGA-binding sites on bovine epididymal spermatozoa: TEM of specimens in toto. Biol Cell 50:279–284
Glabe CG, Grabel LB, Vacquier VD, Rosen SD (1982) Carbohydrate specificity of sea urchin sperm binding: Cell surface lectin mediating sperm-egg adhesion. J Cell Biol 94:123–128
Geoghegan WD, Ackermann GA (1977) Adsorption of horseradish peroxidase, ovomucoid and anti-immunoglobulin to colloidal gold for the indirect detection of Con A WGA and anti-goat anti human immunoglobulin G on cell surfaces at the electron microscopical level: a new method, theory and application. J Histochem Cytochem 25:1187–1200
Hinrichsen-Kohane AC, Hinrichsen M, Schill W-B (1984) Event leading to fertilization — a review. Andrologia 16:321–341
Huang TTF, Yanagimachi R (1984) Fucoidan inhibits attachment of guinea pig spermatozoa to the zona pellucida through binding to the inner acrosomal membrane and equatorial domains. Exp Cell Res 153:363–373
Huang TTF, Ohzu Jr E, Yanagimachi R (1982) Evidence suggesting that L-fucose is part of a recognition signal for sperm-zona pellucida attachment in mammals. Gamete Res 5:355–361
Karnovsky MJ (1965) A formaldehyde-glutaraldehyde fixative of high osmolality for use in electron microscopy. J Cell Biol 27:213–236
Kieda C, Roche AC, Delmotte F, Monsigny M (1979) Lymphocyte membrane lectins. Direct visualization by the use of fluoresceinyl-glycosylated cytochemical markers. FEBS Lett 99:329–331
Koehler JK (1981) Lectins as probes of the spermatozoan surface. Arch Androl 6:197–217
Medcalf DG, Larsen B (1977) Fucose-containing polysaccharides in the brown algae Ascophyllum nodosum and Fucus vesiculosus. Carbohydr Res 59:531–537
Peterson RN, Russell LD, Bundman D, Freund M (1980) Spermegg interaction: direct evidence for boar plasma membrane receptors for porcine zona pellucida. Science 207:73–74
Peterson RN, Russell LD, Bundman D, Conway M, Freund M (1981a) The interaction of living boar sperm and sperm plasma membrane vesicles with the porcine zona pellucida. Dev Biol 84:144–156
Peterson RN, Russell LD, Spaulding G, Bundman D, Buchanan J, Freund M (1981b) Electrophoretic and chromatographic properties of boar sperm plasma membranes: antigens and polypeptides with affinity for isolated zonae pellucida. J Androl 2:300–311
Peterson RN, Russell LD, Hunt WP (1984) Evidence for specific binding of uncapacitated boar spermatozoa to porcine zonae pellucida in vitro. J Exp Zool 231:137–147
Roseman S (1970) The synthesis of complex carbohydrates by multiglycosyltransferase systems and their potential function in intercellular adhesion. Chem Phys Lipids 5:270–297
Shur BD, Hall NG (1982a) Sperm surface galactosyltransferase activity during in vitro capacitation. J Cell Biol 95:567–573
Shur BD, Hall NG (1982b) A role for mouse sperm surface galactosyltransferase in sperm binding to the egg zona pellucida. J Cell Biol 95:574–579
Töpfer-Petersen E, Schill W-B (1983) Characterization of lectin receptors isolated from the outer acrosomal membrane of boar spermatozoa. Int J Androl 6:375–392
Towbin H, Staehelin 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
Vacquier VD, Moy GW (1977) Isolation of binding: the protein responsible for adhesion of sperm to sea urchin eggs. Proc Natl Sci USA 74:2456–2460
Yanagimachi R (1981) Mechanism of fertilization in mammals. In: Mastroinanni L, Biggers JD (eds) Fertilization and embryonic development in vitro. Plenum Press, New York, pp 81
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Töpfer-Petersen, F., Friess, A.E., Nguyen, H. et al. Evidence for a fucose-binding protein in boar spermatozoa. Histochemistry 83, 139–145 (1985). https://doi.org/10.1007/BF00495144
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00495144