Electron microscopic immunolocalization of a conserved sperm acrosomal antigen recognized by HS-63 monoclonal antibody.
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.
Chao
H T
HT
Department of Obstetrics/Gynecology, Taipei Veterans General Hospital, Taiwan, ROC.
Ng
H T
HT
Leng
C H
CH
Lee
C Y
CY
Wei
Y H
YH
D000177:Acrosome / Q000187:drug effects / Q000276:immunology*; D000911:Antibodies, Monoclonal; D000941:Antigens / Q000032:analysis*; D000001:Calcimycin / Q000494:pharmacology; D005455:Fluorescent Antibody Technique; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D008854:Microscopy, Electron; D013081:Sperm Motility
D000911:Antibodies, Monoclonal; D000941:Antigens; D000001:Calcimycin