Center for Genomics and Bioinformatics Research, Division of Biological Science, Faculty of Science, Prince of Songkla University, Hat Yai, Songkhla 90112, Thailand.
National Center for Genetic Engineering and Biotechnology, 113 Thailand Science Park, Phahonyothin Rd., Khlong Luang, Pathum Thani 12120, Thailand.
Comp Biochem Physiol A Mol Integr Physiol. 2021 Nov;261:111045. doi: 10.1016/j.cbpa.2021.111045. Epub 2021 Aug 4.
The long non-coding RNAs (lncRNAs) have been known to play important roles in several biological processes as well as in reproduction. This study aimed to identify lncRNA in ovary female banana shrimp, Fenneropenaeus merguiensis, and investigate the potential role of lncPV13 in the vitellogenesis. After the in silico identification of the ovarian transcriptome, a total of 24,733 putative lncRNAs were obtained, and only 147 putative lncRNAs were significantly differentially expressed among the ovarian development stages. To validate the in silico identification of lncRNAs, the 16 lncRNAs with the highest differential expression in the transcriptome analysis were evaluated by RT-qPCR. The 6 lncRNAs showed higher expression levels in the mature stage than in the previtellogenic stage and were found in several tissues such as in eyestalks, brains, thoracic ganglia, gills, and muscle. Furthermore, most candidate lncRNAs were amplifiable in Litopenaeus vannamei's and Penaeus monodon's DNA but not in Macrobrachium rosenbergii's DNA, suggesting some lncRNAs are expressed in a species-specific manner among penaeid shrimp. In this study, the lncPV13 was investigated for its vitellogenin regulating function by RNA interference. The result indicates that the lncPV13 expression was suppressed in the ovary on day 7 after the injection of double-stranded RNA specific to lncPV13 (dslncPV13), while vitellogenin (Vg) expression was significantly decreased. In contrast, the gonad inhibiting hormone (GIH) expression was significantly increased in the lncPV13 knockdown shrimp. However, the oocyte proliferation was not significantly different between control and lncPV13 knockdown shrimp. This suggests that lncPV13 regulate Vg synthesis through GIH inhibition. Finally, our findings provide lncRNA information and potential lncRNAs involved in the vitellogenesis of female banana shrimp.
长链非编码 RNA(lncRNA)已被证明在多个生物过程以及生殖中发挥重要作用。本研究旨在鉴定雌性香蕉虾卵巢中的 lncRNA,并研究 lncPV13 在卵黄发生中的潜在作用。在卵巢转录组的计算机识别后,共获得了 24733 个推定的 lncRNA,只有 147 个推定的 lncRNA在卵巢发育阶段之间存在显著差异表达。为了验证 lncRNA 的计算机识别,通过 RT-qPCR 评估了转录组分析中差异表达最高的 16 个 lncRNA。这 6 个 lncRNA在成熟阶段的表达水平高于前卵黄发生阶段,并且在眼柄、脑、胸神经节、鳃和肌肉等几种组织中发现。此外,大多数候选 lncRNA 在凡纳滨对虾和斑节对虾的 DNA 中可扩增,但在罗氏沼虾的 DNA 中不可扩增,这表明一些 lncRNA 在对虾中以物种特异性的方式表达。在本研究中,通过 RNA 干扰研究了 lncPV13 对卵黄蛋白原的调节功能。结果表明,在注射双链 RNA 特异性 lncPV13(dslncPV13)后的第 7 天,卵巢中 lncPV13 的表达受到抑制,而卵黄蛋白原(Vg)的表达显著降低。相反,在 lncPV13 敲低虾中,性腺抑制激素(GIH)的表达显著增加。然而,在对照组和 lncPV13 敲低虾之间,卵母细胞增殖没有显著差异。这表明 lncPV13 通过抑制 GIH 调节 Vg 的合成。最后,我们的研究结果提供了 lncRNA 信息和参与雌性香蕉虾卵黄发生的潜在 lncRNA。