ABSTRACT
Artificial insemination with frozen-thawed spermatozoa is commonly used in cattle breeding. A simple and fast procedure is needed for routine evaluation of the acrosomal status of frozen-thawed bovine sperm. Therefore, the purpose of this study was to test two staining procedures used to determine the viability and integrity of acrosome of frozen-thawed bovine spermatozoa. Double staining and Hoechst/FITC-Pisum sativum agglutinin (FITC-PSA) labelling were tested for evaluating the viability and acrosome reaction induced by calcium ionophore of bull spermatozoa. In our experiments no significant differences were detected in the frequency of acrosome-reacted sperm either by double staining (37.98%) or by FITC-PSA labelling (39.33%). The viability of sperm stained by the double staining method was 67.17%, and a higher portion of viable sperm (82.67%) was observed by staining with the Hoechst procedure (P < 0.01). On the basis of the results obtained it is concluded that both methods can be used for detecting the acrosome reaction of frozen-thawed bovine spermatozoa.
Subject(s)
Acrosome/physiology , Cattle/physiology , Spermatozoa/cytology , Staining and Labeling/veterinary , Acrosome Reaction/physiology , Animals , Cell Survival , Freezing , Male , Semen Preservation/veterinary , Staining and Labeling/methodsABSTRACT
The cattle V antigen from the FV blood group system was characterized. Hemolytic as well as immunochemical analyses with monoclonal antibody (MAb) IVA-41 found that V antigen of bovine red blood cells is a membrane-bound, papain- and pronase-sensitive, trypsin- and chymotrypsin-resistant N-glycosylated sialoglyco-protein with molecular weight of 64, 56, and 50 kDa under no reduction and 23 kDa under reduction conditions. In contrary to some human blood group antigens, the expression of bovine blood group V antigen is restricted to the erythrocyte membrane.
Subject(s)
Antibodies, Monoclonal/chemistry , Blood Group Antigens/chemistry , Blood Group Antigens/immunology , Cattle/immunology , Erythrocytes/chemistry , Erythrocytes/immunology , Animals , Antibodies, Monoclonal/biosynthesis , Blood Group Antigens/genetics , Bone Marrow Cells/cytology , Bone Marrow Cells/immunology , Humans , Organ Specificity , Sialoglycoproteins/chemistry , Sialoglycoproteins/immunology , Spleen/cytology , Spleen/immunologyABSTRACT
Monoclonal antibody IVA-285 (IgG1 isotype) recognizing antigenic determinant on bovine and ovine immunoglobulin light chain was produced and characterized. Western blot analysis of bovine immunoglobulin G (IgG) and immunoglobulin M (IgM) purified from bovine blood serum as well as whole immunoglobulin fractions of bovine and ovine serum with IVA-285 showed a molecular weight in the 24-27 kd range corresponding to the Ig light chain of bovine Ig. IVA-285 recognizes the Ig light chain on Ig+ lymphocyte subpopulation and in the majority of body fluids; however, especially strong reactions were observed in bovine tissues (lymph node follicles, plasma cells, and Ig deposits in various tissues).
