Summary
In order to study the acrosome reaction in boar, spermatozoa were incubated in a calcium-containing medium in the presence of the calcium ionophore A23187. The time course of the acrosome reaction was assessed by phasecontrast microscopy and correlated with the movement characteristics of the spermatozoa determined by means of multiple-exposure photography (MEP). Different stages of the acrosome reaction could be observed by indirect immunofluorescence using an antibody fraction raised in rabbits against the isolated outer acrosomal membrane (OAM). At the start of the acrosome reaction, a bright fluorescence located exclusively at the acrosomal cap of the sperm head could be observed, whereas after 60–120 min, the fluorescence vanished, indicating the complete loss of the OAM. However, to gain more insight into the stages of the plasma membrane and OAM during the acrosome reaction, immunoelectron-microscopical studies were performed using anti-OAM antibodies detected by the protein-A gold method. Ultrathin sections and total preparations in combination with transmission electron microscopy (TEM) confirmed, that boar spermatozoa start their acrosome reaction by a vesiculation of the plasma membrane, thus exposing the heavily labelled OAM, which is then lost as sheets or large vesicles. The newly exposed inner acrosomal membrane did not show any labelling with gold, thereby indicating clear differences in the antigenicity of both acrosomal membranes.
Similar content being viewed by others
References
Austin CR (1951) Observation on the penetration of the sperm into the mammalian egg. Aust J Sci Res B4:581–589
Austin CR (1975) Membrane fusion events in fertilization. J Reprod Fert 44:155–166
Austin CR, Bishop MWH (1958) Some features of the acrosome and the perforatorium in mammalian spermatozoa. Proc R Soc Ser B 149:234–240
Avrameas S, Ternynck T (1969) The cross-linking of proteins with glutarealdehyde and its use for the preparation of immunoadsorbents. Immunochemistry 6:53–66
Barros C, Bedford JM, Franklin LE, Austin CR (1967) Membrane fusion as a feature of the mammalian acrosome reaction. J Cell Biol 34:C1-C5
Bedford JM (1969) Morphological aspects of capacitation. In: Raspe (ed) Schering Symposium on mechanism involved in conception. Advances in the Biosciences, vol 4. Pergamon Press, Vieweg, London-Braunschweig, pp 35–50
Bedford JM (1970) Sperm capacitation and fertilization in mammals. Biol Reprod (Suppl) 2:128–158
Bedford JM, Nicander L (1971) Ultrastructural changes in the acrosome and sperm membranes during maturation of the spermatozoa in the testis and epididymis of the rabbit and monkey. J Anat 108:527–543
Brown CR, Harrison RAP (1978) The activation of proacrosin in spermatozoa from ram, bull and boar. Biochim Biophys Acta 526:202–217
Chang MC (1951) Fertilizing capacity of spermatozoa deposited in the fallopian tubes. Nature 168:697–698
Friess AE, Sinowatz F (1984) Con A and WGA binding sites of bovine epididymal spermatozoa. TEM of specimen in toto. Biol Cell 50:279–284
Goodpasture JC, Reddy JM, Zanefeld LJM (1981) Acrosin, proacrosin and acrosin inhibitor of guinea pig spermatozoa capacitated and acrosome-reacted in vitro. Biol Reprod 25: 41–55
Horrisberger M (1981) Colloidal gold: A cytochemical marker for light and fluorescent microscopy and for transmission and scanning electron microscopy. In: O'Hare (ed) Scanning Electron Microscopy 1981/III, pp 9–31
Hyne RV, Garbers DL (1979) Calcium-dependent increase in adenosine-3-5-monophosphate and induction of the acrosome reaction in guinea pig spermatozoa. Proc Natl Acad Sci USA 76:5699–5705
Jones RC (1973) Changes occurring in the head of boar spermatozoa. Vesiculation or vacuolation of the acrosome. J Reprod Fert 33:113–118
Makler A (1978) A new multiple exposure photography method for objective sperm motility determination. Fert Steril 30:192–199
Multamäcki S, Pelliniemi LJ (1973) Ultrastructure of subcellular fractions of bull spermatozoa after hyamine treatment. Z Zellforsch Mikrosk Anat 144:395–408
Ouchterlony Ö (1968) Handbook of immunodiffusion and immunoelectrophoresis. Ann Arbor Science Publishers, Ann Arbor
Peterson R, Russell L, Bundman D, Freund M (1978) Presence of microfilaments and tubular structures in boar spermatozoa after chemically induced acrosome reaction. Biol Reprod 19:459–466
Roosmann GM, Afzelius BA (1975) Acrosome vesiculation in human sperm. J Submicrosc Cytol 7:61–70
Roth J (1982) The protein A-gold (pAg) technique. Qualitative and quantitative approach for antigen localization on thin sections. In: Bullock GR, Petrusz P (eds) Techniques in immunochemistry, vol 1. Academic Press, New York, pp 103–135
Schwarz MA, Koehler JK (1979) Alteration in lectin binding to guinea pig spermatozoa accompanying in vitro capacitation and acrosome reaction. Biol Reprod 21:1295–1307
Shams-Borhan G, Harrison RAP (1981) Production, characterization and use of ionophore-induced, calcium-dependent acrosome reaction in ram spermatozoa. Gamete Res 4:407–432
Sinowatz F, Wrobel K-H (1981) Development of the bovine acrosome. An ultrastructural and cytochemical study. Cell Tissue Res 219:511–524
Töpfer-Petersen E, Schill W-B (1981) A new separation method of subcellular fractions of boar spermatozoa. Andrologia 13:174–176
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
Töpfer-Petersen E, Schmoeckel C, Schill W-B (1983) The acrosomal membrane system-morphological and biochemical studies. Andrologia 15:62–70
Töpfer-Petersen E, Auerbeck J, Weiss A, Friess AE, Schill W-B (1985) The sperm acrosome: Immunological investigations using a specific antiserum and four monoclonal antibodies. Int J Fert (submitted for publication)
Voller A, Bidwell DE, Bartlett A (1979) The enzyme-linked immunoassay (ELISA). Dynatech, Guernsey
Wooding FBP (1973) The effect of Triton X-100 on the ultrastructure of ejaculated bovine sperm. J Ultrastruct Res 42:502–516
Yanagimachi R, Noda YD (1970) Physiologic changes in the post nuclear cap regions of mammalian spermatozoa: a necessary prelimary to the membrane fusion between sperm and egg cells. J Ultrastruct Res 31:486–493
Yanagimachi R, Phillips DM (1984) The status of acrosomal caps of hamster spermatozoa immediately before fertilization in vivo. Gamete Res 9:1–19
Zahler WL, Doak GA (1975) Isolation of the outer acrosomal membrane from bull sperm. Biochim Biophys Acta 406:479–488
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Töpfer-Petersen, E., Friess, A.E., Sinowatz, F. et al. Immunocytological characteriziation of the outer acrosomal membrane (OAM) during acrosome reaction in boar. Histochemistry 82, 113–120 (1985). https://doi.org/10.1007/BF00708194
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00708194