Development and immunity-related microRNAs of the lepidopteran model host Galleria mellonella.
Development and immunity-related microRNAs of the lepidopteran model host Galleria mellonella.
复制标题
DOI：
10.1186/1471-2164-15-705
复制
发表时间：
2014-08-23
期刊：
影响因子：
4.4
通讯作者：
Vilcinskas A
中科院分区：
文献类型：
--
作者：
Mukherjee K;Vilcinskas A
MicroRNAs （miRNAs）是一种小的非编码rna，在蛋白质合成的转录后调控中起着关键作用。虽然我们对它们在发育和免疫等复杂生理过程中的作用知之甚少，但由于模型系统的使用，我们的知识正在迅速扩大。大蜡蛾（Galleria mellonella）的幼虫现在被确定为感染昆虫或人类的病原体的模式宿主。为了建立在我们之前报道的全面的mellonella转录组的基础上，我们在这里描述了发育和免疫相关mirna的鉴定和分析，从而提供了有价值的额外数据，以促进该模型宿主用于复杂过程的分析。为了筛选大黄蜂（G. mellonella）(1)在变态过程中、(2)感染昆虫病原细菌嗜虫塞拉菌（Serratia entom病原细菌）或(3)感染寄生真菌绿僵菌（Metarhizium anisopliae）后差异表达的miRNA，我们设计了一个包含2000多个昆虫miRNA探针序列的微阵列。我们鉴定出的mirna在感染绿僵菌的蛹前（16）、蛹（22）和末龄幼虫(1)中显著表达，并与未处理的末龄幼虫进行比较作为参考。然后，我们使用转录组数据库通过考虑碱基对互补和最小自由能杂交来确定潜在的形成miRNA-mRNA双链的3 '非翻译区域。我们通过RT-PCR证实了选定的miRNAs（如miR-71、miR-263a和miR-263b）与其预测的靶mrna在末龄幼虫、蛹前和蛹中共表达。我们还发现了在细菌或真菌病原体感染时表达的miRNA，以及一个可能作为跨代免疫启动的候选介质的miRNA。这是首次在鳞翅目模型宿主G. mellonella中鉴定出预测在变态或感染反应中调节基因表达的miRNAs的研究。本文的在线版本（doi:10.1186/1471-2164-15-705）包含补充材料，可供授权用户使用。
MicroRNAs (miRNAs) are small non-coding RNAs that act as key players in the post-transcriptional regulation of protein synthesis. Although little is known about their role in complex physiological processes such as development and immunity, our knowledge is expanding rapidly, thanks to the use of model systems. The larvae of the greater wax moth Galleria mellonella are now established as model hosts for pathogens that infect insects or humans. To build on our previously-reported comprehensive G. mellonella transcriptome, here we describe the identification and analysis of development and immunity-related miRNAs, thus providing valuable additional data to promote the use of this model host for the analysis of complex processes. To screen for miRNAs that are differentially expressed in G. mellonella (1) during metamorphosis or (2) following infection with the entomopathogenic bacterium Serratia entomophila or (3) with the parasitic fungus Metarhizium anisopliae, we designed a microarray containing more than 2000 insect miRNA probe sequences. We identified miRNAs that were significantly expressed in pre-pupae (16), pupae (22) and last-instar larvae infected with M. anisopliae (1) in comparison with untreated last-instar larvae which were used as a reference. We then used our transcriptomic database to identify potential 3′ untranslated regions that form miRNA–mRNA duplexes by considering both base pair complementarity and minimum free energy hybridization. We confirmed the co-expression of selected miRNAs (such as miR-71, miR-263a and miR-263b) with their predicted target mRNAs in last-instar larvae, pre-pupae and pupae by RT-PCR. We also identified miRNAs that were expressed in response to infection with bacterial or fungal pathogens, and one miRNA that may act as a candidate mediator of trans-generational immune priming. This is the first study to identify miRNAs that are predicted to regulate genes expressed during metamorphosis or in response to infection in the lepidopteran model host G. mellonella. The online version of this article (doi:10.1186/1471-2164-15-705) contains supplementary material, which is available to authorized users.
登录
查看更多内容
影响因子：
3.3
作者：
Freitak D;Knorr E;Vogel H;Vilcinskas A
通讯作者：
Vilcinskas A
DOI：
10.1073/pnas.0907391106
发表时间：
2009-12-22
影响因子：
11.1
作者：
Gomez-Orte, Eva;Belles, Xavier
通讯作者：
Belles, Xavier
影响因子：
6.7
作者：
Martins NE;Faria VG;Teixeira L;Magalhães S;Sucena É
通讯作者：
Sucena É
影响因子：
64.8
作者：
Maynard, Craig L.;Elson, Charles O.;Hatton, Robin D.;Weaver, Casey T.
通讯作者：
Weaver, Casey T.
影响因子：
2.8
作者：
Mukherjee K;Fischer R;Vilcinskas A
通讯作者：
Vilcinskas A