Characterization of boar spermadhesins by monoclonal and polyclonal antibodies and their role in binding to oocytes.
OBJECTIVE
The role of Ala-Trp-Asn (AWN) and Ala-Gln-Asn (AQN) families of spermadhesive sperm proteins in fertilization.


METHODS
The preparation and characterization of polyclonal antibodies against AWN and AQN spermadhesins and one monoclonal antibody (MAb), designated Bo.5, against AWN spermadhesin. The use of biochemical and immunocytochemical methods for characterization of spermadhesins on the sperm membrane of boar spermatozoa and in the cryostat sections of boar reproductive organs.


RESULTS
Polyclonal anti-AWN and anti-AQN antibodies specifically reacted with AWN and AQN proteins, respectively. MAb Bo.5 detected the 17-, 16-, and 14-kDa protein members of AWN subfamily. The monoclonal, as well as the polyclonal, AWN antibodies remarkably decreased the sperm binding to the egg surface in an in vitro sperm zona pellucida binding assay.


CONCLUSIONS
Presented results demonstrate that polyclonal antibodies and MAb Bo.5 against spermadhesins specifically recognize the membrane-associated antigens and inhibit the binding of sperm to oocytes. Reduced binding of sperm to oocytes, due to the antibodies, indicates the role of these spermadhesins in sperm-egg primary binding.
Veselský
L
L
Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Prague.
Peknicová
J
J
Cechová
D
D
Kraus
M
M
Geussová
G
G
Jonáková
V
V
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism*; D001666:Binding Sites, Antibody; D015815:Cell Adhesion Molecules / Q000276:immunology* / Q000378:metabolism / Q000502:physiology; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect; D006023:Glycoproteins / Q000276:immunology* / Q000302:isolation & purification / Q000502:physiology; D006801:Humans; D008297:Male; D009865:Oocytes / Q000276:immunology / Q000378:metabolism* / Q000502:physiology; D011817:Rabbits; D011956:Receptors, Cell Surface; D013084:Sperm-Ovum Interactions / Q000276:immunology*; D013094:Spermatozoa / Q000276:immunology / Q000378:metabolism / Q000502:physiology*; D013552:Swine; D015044:Zona Pellucida / Q000378:metabolism / Q000502:physiology
D000911:Antibodies, Monoclonal; D015815:Cell Adhesion Molecules; D006023:Glycoproteins; D011956:Receptors, Cell Surface; C035027:egg surface sperm receptor