Laser capture microdissection enables transcriptomic analysis of dividing and quiescent liver stages of Plasmodium relapsing species.
Laser capture microdissection enables transcriptomic analysis of dividing and quiescent liver stages of Plasmodium relapsing species.
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激光捕获显微解剖可以实现疟原虫复发物种的分裂和静止肝阶段的转录组分析。
DOI：
10.1111/cmi.12735
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发表时间：
2017-08
影响因子：
3.4
通讯作者：
Mazier D
中科院分区：
文献类型：
--
作者：
Cubi R;Vembar SS;Biton A;Franetich JF;Bordessoulles M;Sossau D;Zanghi G;Bosson-Vanga H;Benard M;Moreno A;Dereuddre-Bosquet N;Le Grand R;Scherf A;Mazier D
间日疟原虫的休眠肝脏阶段形式(催眠体)是控制和根除感染的主要障碍。尽管进行了大量的研究工作，但催眠体的分子组成仍然不明确。在这里，我们应用了最先进的技术组合来产生第一个催眠虫转录组。我们开发了一种强大的激光解剖显微镜方法，从感染的猴肝细胞中分离食蟹猴疟原虫催眠体和裂殖体，并优化了RNA-SEQ分析，以获得这些阶段的第一个转录本。比较转录学分析发现，有120个转录本在催眠期与肝脏裂殖体分裂期有差异表达，其中69和51个mRNAs在催眠期表达上调或下调。这导致了对承诺和维持催眠动物休眠状态的潜在标志的识别，包括ApiAP2家族的三个转录调节因子，其中一个是食蟹猴和间日疟原虫所特有的，以及全球翻译抑制因子eIF2a激酶eIK2，所有这些都在催眠虫中上调。总之，这项工作不仅提供了一个初步的实验衍生的催眠动物分子标记清单，而且还确定了基因表达的转录和转录后调节可能是建立和维持静止的关键。
Dormant liver stage forms (hypnozoites) of the malaria parasite Plasmodium vivax present major hurdles to control and eradicate infection. Despite major research efforts, the molecular composition of hypnozoites remains ill defined. Here, we applied a combination of state‐of‐the‐art technologies to generate the first transcriptome of hypnozoites. We developed a robust laser dissection microscopy protocol to isolate individual Plasmodium cynomolgi hypnozoites and schizonts from infected monkey hepatocytes and optimized RNA‐seq analysis to obtain the first transcriptomes of these stages. Comparative transcriptomic analysis identified 120 transcripts as being differentially expressed in the hypnozoite stage relative to the dividing liver schizont, with 69 and 51 mRNAs being up‐ or down‐regulated, respectively, in the hypnozoites. This lead to the identification of potential markers of commitment to and maintenance of the dormant state of the hypnozoite including three transcriptional regulators of the ApiAP2 family, one of which is unique to P. cynomolgi and P. vivax, and the global translational repressor, eIF2a kinase eIK2, all of which are upregulated in the hypnozoite. Together, this work not only provides a primary experimentally‐derived list of molecular markers of hypnozoites but also identifies transcriptional and posttranscriptional regulation of gene expression as potentially being key to establishing and maintaining quiescence.
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影响因子：
6.7
作者：
Miao, Jun;Fan, Qi;Cui, Liwang
通讯作者：
Cui, Liwang
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Dembele, Laurent;Franetich, Jean-Francois;Mazier, Dominique
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Mazier, Dominique
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作者：
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通讯作者：
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