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鉴定精子抗原是开发一种避孕疫苗以减少穴居水鼠(Arvicola terrestris Scherman)繁殖数量的第一步。

Identification of sperm antigens as a first step towards the generation of a contraceptive vaccine to decrease fossorial water vole Arvicola terrestris Scherman proliferations.

作者信息

Grignard E, Cadet R, Saez F, Drevet J R, Vernet P

机构信息

Laboratoire Epididyme & Maturation des Gamètes, Université Blaise Pascal, UMR CNRS 6547-GEEM, 24 avenue des Landais, 63177, Aubière Cedex, France.

出版信息

Theriogenology. 2007 Sep 15;68(5):779-95. doi: 10.1016/j.theriogenology.2007.06.010. Epub 2007 Jul 23.

Abstract

Immunocontraceptive strategies have proved to be efficient in controlling fertility of various mammalian species. In the present study we have made the first steps towards the identification of Arvicola terrestris sperm antigens that could be used as targets in the development of a contraceptive vaccine to limit the proliferations of this pest rodent. Rabbit-raised polyclonal antisera directed against complete A. terrestris spermatozoa were used to identify and characterize on 2D-gels coupled with a MALDI-TOF mass spectrometry analysis A. terrestris sperm proteins. Amongst the proteins pinpointed by this approach some were further investigated based on their tissue- and/or sperm-specific expression, and their relevance to fertility or sperm/egg interaction. In parallel, three proteins that have been already reported in the literature to be appropriate targets for the development of contraceptive vaccines in other mammalian species have also been looked for in A. terrestris. With the selected protein targets, a reverse-PCR approach using degenerate primers was employed to amplify corresponding A. terrestris cDNAs. After conceptual translation and sequence alignment, different proteins were studied to determine zones with sufficient sequence divergence and of antigenic/immunogenic nature that could be used in future assays to immunize animals.

摘要

免疫避孕策略已被证明在控制各种哺乳动物的生育能力方面是有效的。在本研究中,我们朝着鉴定水田鼠精子抗原迈出了第一步,这些抗原可作为开发避孕疫苗的靶点,以限制这种有害啮齿动物的繁殖。用兔制备的针对完整水田鼠精子的多克隆抗血清,结合基质辅助激光解吸电离飞行时间质谱分析,在二维凝胶上鉴定和表征水田鼠精子蛋白。在通过这种方法确定的蛋白质中,根据它们的组织和/或精子特异性表达以及它们与生育能力或精子/卵子相互作用的相关性,对其中一些进行了进一步研究。同时,还在水田鼠中寻找了三种已在文献中报道的可作为其他哺乳动物避孕疫苗开发合适靶点的蛋白质。对于选定的蛋白质靶点,使用简并引物通过逆转录PCR方法扩增相应的水田鼠cDNA。经过概念性翻译和序列比对后,研究了不同的蛋白质,以确定具有足够序列差异且具有抗原性/免疫原性的区域,这些区域可用于未来免疫动物的试验。

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