Anti-sperm monoclonal antibodies (mAbs) were evaluated as biomarkers to assess
capacitation, acrosome reaction, cryodamage and fertility of bull spermatozoa in vitro. Three
anti-human sperm mAbs crossreacting with bull sperm antigens were identified. They showed
time-dependent changes in binding to bull spermatozoa incubated under capacitation conditions
when assessed by indirect immunofluorescence, with maximum binding at 4 h. Bull and human
sperm antigens recognized by one mAb (HS-11) were isolated and their immunological
relatedness shown by ELISA. The possible relationship between fertility and the binding of HS-
11 to bull (n=8) spermatozoa was tested in a bovine in vitro fertilization system. In vitro
fertility based on cleavage of oocytes and mAb-binding to spermatozoa at 4 h incubation under
capacitation conditions were correlated (r=0.43; n=32; P<0.05), but in vivo fertility and
sperm-HS-11 binding were not correlated.
In the next set of experiments, 15 mAbs specific to bull sperm antigens were generated.
The mAbs were sperm-specific, but not species-specific. The mAbs identified five distinct
antigenic domains of bull spermatozoa. Seven of the 13 mAbs tested, recognized bull sperm
proteins of >200 kDa in western blots. Three detected more than one protein band (40-200
kDa), while three recognized none.
Thirteen of the 15 mAbs were tested for their influence on bovine sperm-zona
interactions in vitro. Sperm-zona binding was not affected by 12 mAbs. However, sperm-zona
binding in the presence of one surface-reacting mAb was higher than the control (23.6+5.6 vs
10.0+2.4 sperm/zona, mean+SE; P<0.001). Four mAbs specific to intra-acrosomal antigens
exhibited a time dependent increase (P<0.05) in binding to bull spermatozoa incubated under
capacitation conditions. In contrast, the binding of mAbs specific to surface antigens decreased (P < 0.05) after 4 h incubation in the presence of heparin. Following induced acrosome reaction,
a significant decrease (P<0.01) in the binding of acrosome-specific mAbs was observed. Four
selected mAbs were then evaluated as possible indicators of cryodamage in frozen-thawed bull
spermatozoa. Even though the mAbs showed higher binding to frozen-thawed spermatozoa,
there was no conclusive evidence to support the hypothesis that the mAbs will be useful to detect
live membrane-damaged spermatozoa. / Land and Food Systems, Faculty of / Graduate
| Identifer | oai:union.ndltd.org:UBC/oai:circle.library.ubc.ca:2429/6245 |
| Date | 05 1900 |
| Creators | Ambrose, Divakar Justus |
| Source Sets | University of British Columbia |
| Language | English |
| Detected Language | English |
| Type | Text, Thesis/Dissertation |
| Format | 13695485 bytes, application/pdf |
| Rights | For non-commercial purposes only, such as research, private study and education. Additional conditions apply, see Terms of Use https://open.library.ubc.ca/terms_of_use. |
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