Proacrosin/acrosin quantification as an indicator of acrosomal integrity in fresh and frozen dog spermatozoa.
The scope of the present study was to evaluate the presence and activation of proacrosin/acrosin as a tool to determine the acrosomal status of fresh and frozen/thawed dog spermatozoa. Monoclonal antibody C5F11, directed against human acrosin, cross-reacted with dog spermatozoa and labeled the acrosome of both fresh and frozen/thawed dog spermatozoa. Frozen/thawed spermatozoa had a lesser proportion of labeled spermatozoa than fresh spermatozoa (P<0.05). When live spermatozoa were labeled with soybean trypsin inhibitor conjugated with Alexa 488 (SBTI-Alexa 488), the proportion of acrosome-labeled fresh spermatozoa was less than frozen/thawed spermatozoa (P<0.05). By using Western blots and enzymatic activity, frozen/thawed spermatozoa had a greater proportion of active acrosin than fresh spermatozoa. In addition, beta 1,4-galactosyl-transferase (GalT), a plasma membrane bound protein, remained attached to frozen/thawed spermatozoa. Proacrosin is activated during freezing/thawing of dog spermatozoa, and that proacrosin/acrosin may be a good indicator of acrosomal integrity of frozen/thawed spermatozoa.
Cortes
Constanza J
CJ
Departamento de Ciencias Fisiológicas, Facultad de Ciencias Biológicas, Pontificia Universidad Católica de Chile, Alameda 340, Santiago, Región Metropolitana, Chile.
Codelia
Verónica A
VA
Manosalva
Iris
I
de Lange
Johanna
J
De Los Reyes
Mónica
M
Moreno
Ricardo D
RD
D000176:Acrosin / Q000276:immunology* / Q000378:metabolism; D000177:Acrosome / Q000276:immunology / Q000502:physiology; D020101:Acrosome Reaction / Q000502:physiology; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D015153:Blotting, Western / Q000662:veterinary; D004285:Dogs / Q000502:physiology*; D004792:Enzyme Precursors / Q000276:immunology* / Q000378:metabolism; D008297:Male; D012662:Semen Preservation / Q000662:veterinary; D013094:Spermatozoa / Q000201:enzymology / Q000276:immunology / Q000502:physiology*
D000911:Antibodies, Monoclonal; D004792:Enzyme Precursors; C022438:proacrosin; D000176:Acrosin