A characterization of sperm surface proteins using monoclonal antibodies.\n\n\n

0474845 - BTÚ 2018 CZ eng A - Abstrakt
Pohlová, Alžběta - Zigo, Michal - Dorosh, Andriy - Jonáková, Věra - Dvořáková-Hortová, Kateřina - Postlerová, Pavla
A characterization of sperm surface proteins using monoclonal antibodies.


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Book of abstracts XXIIIrd Symposium of Immunology and Biology of Reproduction. Vestec u Prahy: Biotechnologický ústav AVČR v. v. i., 2017 - (Kubátová, a.). s. 27-27
[XXIIIrd Symposium of Immunology and Biology of Reproduction. 18.05.2017-20.05.2017, Třešť]
Grant CEP: GA MŠMT(CZ) ED1.1.00/02.0109; GA AV ČR SAV-AVČR 15-05
Institucionální podpora: RVO:86652036
Klíčová slova: monoclonal antibodies * sperm-zona pellucida receptors * ZP glycoproteins * spermatozoa * capacitation * sperm surface proteins
Obor OECD: Biochemistry and molecular biology

Reproduction is a crucial biological process, which leads to a production of new individuals by combining genetic material of two parental organisms. Proteins are a substantial equipment of the spermatic cell and their characterization can help us to find potential diagnostic markers or identify species-specific sperm-zona pellucida (ZP) receptors. Thus, a unique panel of monoclonal antibodies against boar sperm surface proteins was prepared by immunization of mice. Firstly, an indirect IF technique was used to screen a protein localization on a pig spermatozoa (epididymal, ejaculated and in vitro capacitated). Secondly, proteins isolated from the sperm surface of epididymal, ejaculated and in vitro capacitated sperm were separated by SDS-electrophoresis and used for antibody detection (Western blot) and a binding study with lectin-labeled ZP glycoproteins (Far Western blot). In the pig model, three proteins, which were recognized by monoclonal antibodies and interacted with porcine ZP glycoproteins, were immunoprecipitated and identified as acrosin precursor, RAB-2A protein, and lactadherin P47. Additionally, we tested the whole panel of 17 monoclonal antibodies with the sperm of other mammalian species, such as mouse, bull and human. In these species we found by using an IF technique that a few antibodies recognized proteins localized in the sperm acrosomal region. Antibodies named 1E3, 2E1 and 3B10 recognized proteins on the mouse epididymal sperm and 4C7 and 1E3 antibodies bound to proteins on the bull sperm. Further, we observed a cross-reactivity of these monoclonal antibodies in protein extracts of bull, mouse and human sperm. Especially, antibodies 4C7, 5C5, 1E3 and 2E1 detected proteins in extracts from bull sperm. Preselected antibodies could be used for detection of the sperm-ZP binding receptors in patients with normal and pathological spermiograms and serve as a tool for searching diagnostic fertility markers in the human.







Trvalý link: http://hdl.handle.net/11104/0273884