Monoclonal Antibodies Reveal Novel Localization of SPAG11E in Spermatozoa and the Antifertility Potential of SPAG11E Motifs  

作　　者：Zhi-Kai Wang Yi-Ting Yang Xin-Yu Chen Shuang-Gang Hu Ping Zhu Wan-Xiang Xu Li Ma He-Guo Yu Hua Diao Yong-Lian Zhang 

机构地区：[1]Clinical Laboratory(Hospital of the SIPPR)and NHC Key Laboratory of Reproduction Regulation,Shanghai Institute of Planned Parenthood Research,Shanghai 200032,China [2]Core Facility of Basic Medical Sciences,School of Medicine,Shanghai Jiao Tong University,Shanghai 200025,China [3]Shanghai Key Laboratory of Molecular Andrology,State Key Laboratory of Molecular Biology,Institute of Biochemistry and Cell Biology,Chinese Academy of Sciences,Shanghai 200031,China

出　　处：《Reproductive and Developmental Medicine》2018年第4期191-200,I0001-I0005,共15页生殖与发育医学（英文版）

基　　金：supported by grants from the Natural Science Foundation of China(No.81671508);the Innovation-oriented Science and Technology Grant from NPFPC Key Laboratory of Reproduction Regulation(No.CX2017-01).

摘　　要：Objective: SPAG11E is the first β-defensin that has been reported to activate Ca;uptake and sperm motility. However, the exact subcellular localization and interaction of SPAG11E with sperm remain controversial because of the lack of qualified antibody tools. SPAG11E is also a potential male antifertility target because SPAG11E fragment conjugated with a carrier protein exhibits male contraceptive vaccine potential.However, the fine B-cell epitope motifs of SPAG11E have not been analyzed, which hampered further exploration of the potential target.Methods: Polyclonal and monoclonal antibodies（mcAbs） of mature SPAG11E were raised and qualified with Western blotting. Subcellular localization of SPAG11E was revealed by Western blotting, immunohistochemistry staining, and electron microscopy. B-cell epitopes of rat SPAG11E were mapped by Western blotting using polyclonal and mcAbs. Based on the conservation of the identified epitope motifs between rat and mouse SPAG11E, antifertility potential of the epitope motifs was evaluated by the offspring of the males compromised with specific mcAbs.Results: SPAG11E antibodies of high quality were obtained and all B-cell epitope motifs of rat SPAG 11 E were mapped, in which conserved epitope motifs of SPAG11E in various species were discovered. The epitope motifs recognized by mcAbs were identified respectively. With mcAbs, rat SPAG11E was proved to be expressed in the caput region of epididymis. A novel finding was that SPAG 11 E was located in the flagella and nuclei of sperm as revealed by immunoelectron microscopy. In addition, the males treated with mcAbs（3#-1 and 10#B4） showed apparently fewer offspring.Conclusions: SPAG11E revealed a β-defensin with novel localization in sperm flagellum and nucleus with qualified antibodies. All B-cell epitope motifs of rat SPAG11E were determined, and the antifertility potential was proved by corresponding mcAbs.

关 键 词：Antibody CONTRACEPTION EPITOPE SPAG11E Sperm 

分 类 号：R169.41[医药卫生—公共卫生与预防医学]