长链非编码RNA(lncRNAs)是各种生物过程中的关键调节剂,如基因表达,发展,免疫防御。然而,关于lncRNAs在亚洲蜜蜂(Apiscerana)幼虫肠道发育中的功能知之甚少。这里,根据我们先前从4-获得的深度测序数据,5-,和6天大的A.cerana工人的幼虫肠道(Ac4,Ac5和Ac6组),进行了深入的全转录组研究,以破译表达模式,监管方式,和lncRNAs在A.cerana工幼虫肠道发育过程中的潜在作用,然后验证差异表达的lncRNAs(DElncRNAs)的相对表达和竞争性内源性RNA(ceRNA)轴内的靶向关系。在Ac4vs.Ac5和Ac5vs.Ac6比较组,527和498DElncRNAs被鉴定,分别,这表明lncRNAs在幼虫肠道发育过程中的动态表达。顺式作用分析表明,上述DElncRNAs的330个和393个相邻基因分别涉及29个和32个功能项,例如细胞过程和代谢过程;这些相邻基因也分别参与246和246个途径,例如Hedgehog信号途径和Wnt信号途径。此外,发现79和76个DElncRNAs作为潜在的反义lncRNAs,分别,与72和60个有义链mRNA相互作用。对竞争性内源性RNA(ceRNA)网络的研究表明,Ac4中的75(155)DElncRNA与Ac5(Ac5vs.Ac6)对比组可以靶向7(5)个DEmiRNA,并进一步结合334(248)个DEmRNA,可以注释33(29)个功能术语和186(210)个途径,包括12(16)个细胞免疫和体液免疫途径(溶酶体途径,坏死,MAPK信号通路,等。)和11(10)个发育相关信号通路(Wnt,河马,AMPK,等。).5个随机选择的DElncRNA的RT-qPCR检测证实了所用测序数据的可靠性。此外,双荧光素酶报告基因测定的结果表明,MSCRG.11294.1与miR-6001-y之间以及miR-6001-y与ncbi_107992440之间存在结合关系.这些结果表明,DElncRNAs可能通过调节源基因的转录来调节幼虫肠道的发育过程,与mRNA的相互作用,和ceRNA网络。我们的发现不仅对A.cerana幼虫肠道的发育机制产生了新的见解,但也为进一步的功能解剖提供了候选ceRNA轴。
Long non-coding RNAs (lncRNAs) are crucial modulators in a variety of biological processes, such as gene expression, development, and immune defense. However, little is known about the function of lncRNAs in the development of Asian honey bee (Apis cerana) larval guts. Here, on the basis of our previously obtained deep-sequencing data from the 4-, 5-, and 6-day-old larval guts of A. cerana workers (Ac4, Ac5, and Ac6 groups), an in-depth transcriptome-wide investigation was conducted to decipher the expression pattern, regulatory manners, and potential roles of lncRNAs during the developmental process of A. cerana worker larval guts, followed by the verification of the relative expression of differentially expressed lncRNAs (DElncRNAs) and the targeting relationships within a competing endogenous RNA (ceRNA) axis. In the Ac4 vs. Ac5 and Ac5 vs. Ac6 comparison groups, 527 and 498 DElncRNAs were identified, respectively, which is suggestive of the dynamic expression of lncRNAs during the developmental process of larval guts. A cis-acting analysis showed that 330 and 393 neighboring genes of the aforementioned DElncRNAs were respectively involved in 29 and 32 functional terms, such as cellular processes and metabolic processes; these neighboring genes were also respectively engaged in 246 and 246 pathways such as the Hedgehog signaling pathway and the Wnt signaling pathway. Additionally, it was found that 79 and 76 DElncRNAs as potential antisense lncRNAs may, respectively, interact with 72 and 60 sense-strand mRNAs. An investigation of competing endogenous RNA (ceRNA) networks suggested that 75 (155) DElncRNAs in the Ac4 vs. Ac5 (Ac5 vs. Ac6) comparison group could target 7 (5) DEmiRNAs and further bind to 334 (248) DEmRNAs, which can be annotated to 33 (29) functional terms and 186 (210) pathways, including 12 (16) cellular- and humoral-immune pathways (lysosome pathway, necroptosis, MAPK signaling pathway, etc.) and 11 (10) development-associated signaling pathways (Wnt, Hippo, AMPK, etc.). The RT-qPCR detection of five randomly selected DElncRNAs confirmed the reliability of the used sequencing data. Moreover, the results of a dual-luciferase reporter assay were indicative of the binding relationship between MSTRG.11294.1 and miR-6001-y and between miR-6001-y and ncbi_107992440. These results demonstrate that DElncRNAs are likely to modulate the developmental process of larval guts via the regulation of the source genes\' transcription, interaction with mRNAs, and ceRNA networks. Our findings not only yield new insights into the developmental mechanism underlying A. cerana larval guts, but also provide a candidate ceRNA axis for further functional dissection.