首页 | 本学科首页   官方微博 | 高级检索  
   检索      


Immunological approach to the characterization of the outer acrosomal membrane of boar spermatozoa
Authors:Adriana C Hinrichsen  Edda Tpfer-Petersen  Thomas Dietl  Christian Schmoeckel  Wolf-B Schill
Institution:Adriana C. Hinrichsen,Edda Töpfer-Petersen,Thomas Dietl,Christian Schmoeckel,Wolf-B. Schill
Abstract:Antiserum to purified boar spermatozoan outer acrosomal membrane (OAM) was raised in rabbits and adsorbed with boar liver and serum glutaraldehyde cross-linked immunoadsorbents. The IgG fraction of the antiserum was purified by (NH4)2SO4 precipitation followed by ion-exchange chromatography. Indirect immunofluorescence showed bright fluorescent staining of the acrosomal cap of boar spermatozoa and to a lesser extent of the acrosomes of bull and goat spermatozoa after incubation with anti-OAM-IgG. Immuno-electron microscopy further confirmed the specificity of the antibody for the OAM. Preincubation of the anti-OAM-IgG with isolated OAM, completely abolished its reactivity. When tested by ELISA, anti-OAM-IgG reacted with boar, bull, goat, and human spermatozoa; however, its binding activity to boar spermatozoa was significantly greater as compared to spermatozoa from the other species tested. In an effort to identify OAM antigens recognized by this antiserum, the isolated boar OAM was labeled either with 3H or with 125I and solubilized by mild detergent treatment. The extracted components were immunoprecipitated with anti-OAM-IgG and protein A-bearing S. aureus and the thus isolated antigens were analysed on SDS-PAGE. The results suggest that anti-OAM-IgG recognized one high molecular 3H-labeled glycoprotein (270 kd), and four 125I-labeled polypeptides of lower molecular weight of the boar OAM.
Keywords:spermatozoa  acrosomal membrane  membrane antigens
设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号