Identification of post-vasectomy sperm auto-antigens in fox (Vulpes vulpes) by two-dimensional gel electrophoresis and Western blotting.
The aim of this work was to identify antigenic surface proteins on fox spermatozoa. Six foxes were inguitinally vasectomised, and the time course of antibody response in the sera was studied. Five out of the six foxes reacted to vasectomy with a production of antisperm antibodies. The number of bands recognised by Western blot was maximal 120-150 days after the vasectomy, at the end of the reproductive season. On the whole, 30 bands were recognised between 9 and 150 kDa. The pattern of recognised proteins varied from one fox to another. The humoral response was studied in one fox 2 years after the vasectomy, before, during and after the breeding season. The same proteins were recognised, but the intensity of staining was increased during the testis regression. Using FITC-labelling on sperm smears with fox sera, antigens were localised at or near the sperm surface, either on the acrosome or both on the acrosome and on the flagellum. Using two-dimensional gel electrophoresis and Western blotting, we identified eight areas containing major antigens, recognised by at least two sera. The molecular weights (kDa) and isoelectric points of the repeated antigens were, respectively [150, 6.6-6.0]; [105-98, 6.0-5.5]; [97, 4.6-4.3]; [95, 5.0]; [85-80, 5.4-5.1]; [42, 5.0-4.8]; [17-15, 6.5-5.9]; and [17-15, 5.5-4.8]. The results of this study can be used to characterise more precisely fox sperm auto-antigens by microsequencing the selected proteins.
Verdier
Yann
Y
AFSSA Nancy, Unit of Wildlife Health and Management, BP9, F-54220, Malzeville, France.
Chaffaux
Stéphane
S
Boué
Franck
F
D000818:Animals; D001324:Autoantigens / Q000032:analysis* / Q000502:physiology; D015153:Blotting, Western; D015180:Electrophoresis, Gel, Two-Dimensional; D005589:Foxes; D008297:Male; D013094:Spermatozoa / Q000276:immunology*; D014659:Vasectomy
D001324:Autoantigens