SALUVET, Animal Health Department, Faculty of Veterinary Sciences, Complutense University of Madrid, Ciudad Universitaria s/n, 28040 Madrid, Spain.
J Proteomics. 2012 Apr 18;75(8):2306-18. doi: 10.1016/j.jprot.2012.01.039. Epub 2012 Feb 10.
Neospora caninum is a cyst-forming parasite that has been recognised worldwide as a cause of cattle abortion and neuromuscular disease in dogs. Variations in genetic profiles, behaviour in vitro, and pathogenicity have been established among N. caninum isolates. However, it is unclear which parasite factors are implicated in this intra-specific diversity. Comparative analysis of protein expression patterns may define the determinants of biological diversity in N. caninum. Using DIGE and MALDI-TOF MS techniques, we quantified and identified differentially expressed proteins in the tachyzoite stage across three N. caninum isolates: the virulent Nc-Liv and Nc-Spain 7 isolates, and the attenuated Nc-Spain 1H isolate. Comparison between Nc-Spain 7 and Nc-Spain 1H extracts revealed 39 protein spots that were more abundant in Nc-Spain 7 and 21 in Nc-Spain 1H. Twenty-four spots were also increased in Nc-Spain 7 and 12 in Nc-Liv. Three protein spots were more abundant in the Nc-Liv extracts than in the Nc-Spain 1H extracts. MS analysis identified 11 proteins differentially expressed that are potentially involved in gliding motility and the lytic cycle of the parasite, and oxidative stress. These differences could help to explain variations in behaviour between isolates and provide a better knowledge of mechanisms associated with virulence.
刚地弓形虫是一种形成包囊的寄生虫，已被全球公认为导致牛流产和犬神经肌肉疾病的病原体。不同的刚地弓形虫分离株在遗传特征、体外行为和致病性方面存在差异。然而，目前尚不清楚哪些寄生虫因素与这种种内多样性有关。蛋白质表达谱的比较分析可能可以确定刚地弓形虫生物学多样性的决定因素。本研究使用 DIGE 和 MALDI-TOF MS 技术，对三种刚地弓形虫分离株（毒力株 Nc-Liv 和 Nc-Spain 7 以及弱毒株 Nc-Spain 1H）的速殖子阶段进行了定量和差异表达蛋白分析。Nc-Spain 7 和 Nc-Spain 1H 提取物之间的比较显示，Nc-Spain 7 中有 39 个蛋白斑点丰度更高，Nc-Spain 1H 中有 21 个蛋白斑点丰度更高。Nc-Spain 7 中也有 24 个斑点增加，Nc-Liv 中有 12 个斑点增加。Nc-Liv 提取物中有 3 个蛋白斑点比 Nc-Spain 1H 提取物中的更丰富。MS 分析鉴定出 11 种差异表达的蛋白，这些蛋白可能参与虫体的滑行运动和裂解周期，以及氧化应激。这些差异可能有助于解释分离株之间行为的变化，并提供与毒力相关的机制的更好的认识。
