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猫精子阳离子通道蛋白1(Catsper1)DNA疫苗的构建及其对雄性小鼠的抗生育作用。

Construction of a catsper1 DNA vaccine and its antifertility effect on male mice.

作者信息

Yu Qiong, Mei Xiao-Qin, Ding Xiao-Fang, Dong Ting-Ting, Dong Wei-Wei, Li Hong-Gang

机构信息

Family Planning Research Institute/Center of Reproductive Medicine, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.

Centre of Reproductive Medicine, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.

出版信息

PLoS One. 2015 May 18;10(5):e0127508. doi: 10.1371/journal.pone.0127508. eCollection 2015.

Abstract

Cation channel of sperm 1 (CATSPER1) is a unique sperm cation channel protein, and essential for sperm function and male fertility. CATSPER1 exclusively expresses in meiotic and postmeiotic spermatogenic cells, thus belongs to the spermatogenesis-specific antigen that escape central tolerance. We have previously demonstrated the immunocontraceptive potential of its transmembrane domains and pore region, and reported the antifertility effects of its B-cell epitopes on male mice. Aiming to develop DNA vaccine targeting CATSPER1 for male contraception, here the whole open reading frame of mouse Catsper1 was cloned into the plasmid pEGFP-N1 to obtain a DNA vaccine pEGFP-N1-Catsper1. The vaccine was confirmed to be transcribed and translated in mouse N2a cell in vitro and mouse muscle tissue in vivo. Intramuscular injection with the vaccine on male mice induced specific immune reaction and caused significant inhibition on sperm hyperactivated motility and progressive motility (P<0.001 for both), and consequently reduced male fertility. The fertility rate of experimental group was 40.9%, which was significant lower (P=0.012) than control group (81.8%). No significant change in mating behavior, sperm production and histology of testis/epididymis was observed. Given that Catsper1 exhibits a high degree of homology among different species, Catsper1 DNA vaccine might be a good strategy for developing an immunocontraceptive vaccine for human and animal use.

摘要

精子阳离子通道1(CATSPER1)是一种独特的精子阳离子通道蛋白,对精子功能和男性生育能力至关重要。CATSPER1仅在减数分裂和减数分裂后的生精细胞中表达,因此属于逃避中枢耐受的精子发生特异性抗原。我们之前已经证明了其跨膜结构域和孔区域的免疫避孕潜力,并报道了其B细胞表位对雄性小鼠的抗生育作用。为了开发针对CATSPER1的DNA疫苗用于男性避孕,在此将小鼠Catsper1的整个开放阅读框克隆到质粒pEGFP-N1中,以获得DNA疫苗pEGFP-N1-Catsper1。该疫苗在体外小鼠N2a细胞和体内小鼠肌肉组织中均被证实可转录和翻译。对雄性小鼠进行肌肉注射该疫苗可诱导特异性免疫反应,并对精子超活化运动和渐进性运动产生显著抑制作用(两者均P<0.001),从而降低雄性生育能力。实验组的生育率为40.9%,显著低于对照组(81.8%,P=0.012)。未观察到交配行为、精子产生以及睾丸/附睾组织学的显著变化。鉴于Catsper1在不同物种间具有高度同源性,Catsper1 DNA疫苗可能是开发用于人类和动物的免疫避孕疫苗的良好策略。

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