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Electron microscopic immunolocalization of a conserved sperm acrosomal antigen recognized by HS‐63 monoclonal antibody
Author(s) -
Chao H. T.,
Ng H. T.,
Leng G. H.,
Lee C. Y. G.,
Wei Y. H.
Publication year - 2009
Publication title -
andrologia
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.633
H-Index - 59
eISSN - 1439-0272
pISSN - 0303-4569
DOI - 10.1111/j.1439-0272.1993.tb02710.x
Subject(s) - immunogold labelling , acrosome , sperm , immunofluorescence , monoclonal antibody , antigen , acrosome reaction , chemistry , microbiology and biotechnology , biology , ultrastructure , antibody , andrology , immunology , anatomy , genetics , medicine
Summary. HS‐63 monoclonal antibody was shown to react with a sperm‐specific acrosomal antigen from spermatozoa of a variety of mammalian species. Indirect immunofluorescence showed that HS‐63 was associated with acrosome‐intact sperm after sperm had been fixed with methanol. Electron microscopy (EM) was employed to determine the ultrastructural localization of this sperm antigen. When the immunogold labelled goat anti‐mouse IgG was used as a probe, we demonstrated that HS‐63 monoclonal antibody did not bind to the freshly prepared human spermatozoa. However, gold particles were observed in the intra‐acrosomal region, when the spermatozoa had been pre‐treated with 0.5% Triton X‐100 prior to incubation with HS‐63. We further observed that the immunogold did not stain the inner acrosome membrane when the spermatozoa became acrosome‐reacted. A good correlation was obtained between the percentage of spermatozoa which did not react with HS‐63 as determined by indirect immunofluorescence assay and that of acrosome‐reacted spermatozoa as quantitated by electron microscopy. The results of this EM study were consistent with those obtained by indirect immunofluorescence assay and both indicated that HS‐63 reacts only with the capacitated and acrosome‐intact spermatozoa. Therefore, HS‐63 monoclonal antibody is a useful probe for rapid evaluation of acrosomal status in human spermatozoa.