Genome-wide expression profiling of in vivo-derived bloodstream parasite stages and dynamic analysis of mRNA alterations during synchronous differentiation in Trypanosoma brucei
Genome-wide expression profiling of in vivo-derived bloodstream parasite stages and dynamic analysis of mRNA alterations during synchronous differentiation in Trypanosoma brucei
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DOI：
10.1186/1471-2164-10-427
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发表时间：
2009-09-11
期刊：
影响因子：
4.4
通讯作者：
Matthews, Keith
中科院分区：
文献类型：
--
作者：
Kabani, Sarah;Fenn, Katelyn;Matthews, Keith
背景：锥虫在其复杂的生命周期中经历了广泛的发育变化。其中至关重要的是细长和粗短的血流形式之间的过渡，此后，从粗短的舌蝇中肠前循环形式的分化。这些发育事件是高度调节，时间上可重复的，并伴随着几乎完全在转录后水平介导的表达变化。结果：在这项研究中，我们已经研究了，通过全基因组微阵列分析，基因的mRNA丰度在细长和矮胖形式的T。1细胞，并且也在其同步分化为原周期形式。总之，测定了代表源自五个单独小鼠感染的匹配寄生虫群体的分化的五个生物学重复，其中RNA在其同步分化为前循环形式的时间过程期间的关键生物学时间点衍生。重要的是，这些mRNA谱的生物学背景是通过测定每个群体中同时发生的细胞事件来建立的（表面抗原交换、形态学重构、细胞周期再进入），从而将观察到的基因表达变化与锥虫分化的良好建立的框架联系起来。使用严格的统计学分析和对实验预测的基因表达和表型变化的衍生谱的验证，我们已经建立了在这些重要的生命周期转变过程中受调控的基因表达谱。矮桩和原轮形式之间分化的高度同步性也意味着这些mRNA谱的研究与这种特征良好的发育过渡期间单个细胞内mRNA丰度的变化直接相关。
Background: Trypanosomes undergo extensive developmental changes during their complex life cycle. Crucial among these is the transition between slender and stumpy bloodstream forms and, thereafter, the differentiation from stumpy to tsetse-midgut procyclic forms. These developmental events are highly regulated, temporally reproducible and accompanied by expression changes mediated almost exclusively at the post-transcriptional level.Results: In this study we have examined, by whole-genome microarray analysis, the mRNA abundance of genes in slender and stumpy forms of T. brucei AnTat1.1 cells, and also during their synchronous differentiation to procyclic forms. In total, five biological replicates representing the differentiation of matched parasite populations derived from five individual mouse infections were assayed, with RNAs being derived at key biological time points during the time course of their synchronous differentiation to procyclic forms. Importantly, the biological context of these mRNA profiles was established by assaying the coincident cellular events in each population (surface antigen exchange, morphological restructuring, cell cycle re-entry), thereby linking the observed gene expression changes to the well-established framework of trypanosome differentiation.Conclusion: Using stringent statistical analysis and validation of the derived profiles against experimentally-predicted gene expression and phenotypic changes, we have established the profile of regulated gene expression during these important life-cycle transitions. The highly synchronous nature of differentiation between stumpy and procyclic forms also means that these studies of mRNA profiles are directly relevant to the changes in mRNA abundance within individual cells during this well-characterised developmental transition.