Laboratório de Biologia Molecular de Cestódeos, Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul, Av Bento Gonçalves, 9500, Bloco IV, Prédio 43-421, Sala 206, 91501-970 Porto Alegre, Rio Grande do Sul, Brazil.
Parasitol Res. 2011 Mar;108(3):645-56. doi: 10.1007/s00436-010-2109-2. Epub 2010 Oct 15.
Despite the fact that cestodes represent major etiological agents of both human and domestic animal diseases, little is known about the molecular aspects of cestode development. In this work, Mesocestoides corti, a model cestode species, was studied from the early development of its larval form (tetrathyridium) into adult worms (strobilation) using different proteomic approaches. The protein profiles of M. corti tetrathyridia induced or not induced to undergo strobilation were compared. Proteomic mapping by two-dimensional gel electrophoresis showed the resolution of 248 and 154 spots from tetrathyridia that were subjected or not subjected to strobilation induction, respectively, allowing for the detection of at least nine spots exclusive to each group. Spot analysis by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) or MALDI-TOF MS/MS identified four reference proteins (six spots). LC-MS/MS analyses of protein extracts identified 66 proteins, eight of which were found exclusively in non-induced tetrathyridia, while 13 were found exclusively in strobilation-induced tetrathyridia. Among the proteins exclusively identified in strobilation-induced worms, there was a predominance of proteins with functions relating to chaperone activity and protein synthesis and turnover. Quantitative differential expression analysis between M. corti tetrathyridia prior to and after strobilation induction revealed six proteins upregulated in strobilation-induced worms; these proteins were involved in metabolic pathways, cell proliferation, and cytoskeletal rearrangement. Overall, despite the absence of a sequenced M. corti genome, using sequences from other platyhelminthes, we were able to establish comprehensive protein profiles for tetrathyridia prior to and after strobilation induction and identify several proteins potentially involved in the early events leading to strobilation.
尽管绦虫是人类和家畜疾病的主要病原体,但对于绦虫发育的分子方面知之甚少。在这项工作中,使用不同的蛋白质组学方法研究了 Mesocestoides corti,一种模式绦虫物种,从其幼虫形式(四叶期)向成虫(出尾)的早期发育。比较了诱导或不诱导出尾的 M. corti 四叶期的蛋白质图谱。二维凝胶电泳的蛋白质图谱显示,分别经历或不经历出尾诱导的四叶期的分辨率为 248 和 154 个斑点,从而可以检测到至少 9 个每个组特有的斑点。基质辅助激光解吸/电离飞行时间(MALDI-TOF)质谱(MS)或 MALDI-TOF MS/MS 的斑点分析鉴定了 4 种参考蛋白(6 个斑点)。蛋白质提取物的 LC-MS/MS 分析鉴定了 66 种蛋白质,其中 8 种仅存在于未诱导的四叶期,而 13 种仅存在于出尾诱导的四叶期。在仅在出尾诱导的蠕虫中鉴定的蛋白质中,存在大量与伴侣活性和蛋白质合成和周转相关的功能的蛋白质。在出尾诱导前后的 M. corti 四叶期的定量差异表达分析显示,在出尾诱导的蠕虫中上调了 6 种蛋白质;这些蛋白质参与代谢途径、细胞增殖和细胞骨架重排。总体而言,尽管缺乏已测序的 M. corti 基因组,但使用来自其他扁形动物的序列,我们能够建立出尾诱导前后四叶期的综合蛋白质图谱,并鉴定出几种可能参与出尾早期事件的蛋白质。