Midlej V, Vilela R, Dias A B, Benchimol M
Universidade Santa Ursula, Rio de Janeiro, Brazil.
Vet Parasitol. 2009 Nov 12;165(3-4):216-30. doi: 10.1016/j.vetpar.2009.07.021. Epub 2009 Jul 21.
Tritrichomonas foetus is an extracellular parasite of the reproductive tract in cattle. To investigate the cytopathic effects of T. foetus in deeper parts of the reproductive tract, a bovine primary oviduct epithelial cell system (BOECs) was developed. Reproductive tracts were obtained from cows and the effect of co-incubating T. foetus with BOECs was analyzed by scanning electron, transmission electron and fluorescence microscopy. Viability tests were performed using colorimetric methods, TUNEL (Terminal deoxynucleotidyltransferase-mediated dUTP nick-end labeling), fluorescein diacetate, propidium iodide, JC-1 and annexin-V. The results demonstrate that: (1) the in vitro oviduct epithelium is useful for interaction experiments with T. foetus; (2) T. foetus adheres to the BOECs as single separate cells, and later on the cells aggregate as large clusters; (3) the posterior region of the cell initiates the process of adhesion and forms filopodia and digitopodia; (4) T. foetus severely damages BOECs leaving imprints in the epithelial cells, wide intercellular spaces, and large lesions in the epithelium; and (5) T. foetus provokes bovine oviduct cell death by apoptosis and secondary necrosis. Our observations indicate the possibility that T. foetus can move through the reproductive tract to the oviduct and that infertility in cows can be mediated by an attack on the oviduct cells by T. foetus.
胎儿三毛滴虫是牛生殖道的一种细胞外寄生虫。为了研究胎儿三毛滴虫在生殖道深部的细胞病变效应,建立了牛原代输卵管上皮细胞系统(BOECs)。从奶牛获取生殖道组织,通过扫描电子显微镜、透射电子显微镜和荧光显微镜分析胎儿三毛滴虫与BOECs共同孵育的效应。使用比色法、TUNEL(末端脱氧核苷酸转移酶介导的dUTP缺口末端标记)、荧光素二乙酸酯、碘化丙啶、JC-1和膜联蛋白-V进行活力测试。结果表明:(1)体外输卵管上皮可用于与胎儿三毛滴虫的相互作用实验;(2)胎儿三毛滴虫以单个分离细胞的形式黏附于BOECs,随后细胞聚集成大的团块;(3)细胞后部启动黏附过程并形成丝状伪足和指状伪足;(4)胎儿三毛滴虫严重损伤BOECs,在上皮细胞中留下印记、细胞间隙变宽以及上皮出现大的病变;(5)胎儿三毛滴虫通过凋亡和继发性坏死引发牛输卵管细胞死亡。我们的观察结果表明,胎儿三毛滴虫有可能穿过生殖道到达输卵管,并且奶牛的不育可能是由胎儿三毛滴虫对输卵管细胞的攻击所介导的。