College of Animal Science & Technology, Gansu Agricultural University, Lanzhou 730070, PR China.
College of Animal Science & Technology, Gansu Agricultural University, Lanzhou 730070, PR China.
Int J Biol Macromol. 2023 Jun 1;239:124246. doi: 10.1016/j.ijbiomac.2023.124246. Epub 2023 Mar 31.
Rainbow trout (Oncorhynchus mykiss) is a typical cold-water fish. With global warming and extreme heat, high summer temperatures are the biggest threat to rainbow trout farming. Rainbow trout initiate stress defense mechanisms in response to thermal stimuli, and competing endogenous RNA (ceRNA) regulation of target genes (mRNAs) mediated by non-coding RNAs (microRNAs [miRNAs], long non-coding RNAs) may be the main strategy for responding to thermal stimuli and enhancing adaptation.
We screened the LOC110485411-novel-m0007-5p-hsp90ab1 ceRNA relationship pairs for affect heat stress in rainbow trout and validated their targeting relationships and functions based on preliminary high-throughput sequencing analysis results. The transfection of exogenous novel-m0007-5p mimics and inhibitors into primary rainbow trout hepatocytes effectively bound and inhibited the target genes hsp90ab1 and LOC110485411 without significant effects on hepatocyte viability, proliferation, and apoptosis. The inhibitory effect of novel-m0007-5p overexpression on hsp90ab1 and LOC110485411 under heat stress was time-efficient. Similarly, small interfering RNAs (siRNAs) affected hsp90ab1 mRNA expression by silencing LOC110485411 expression time-efficiently.
In conclusion, we found that in rainbow trout, LOC110485411 and hsp90ab1 can bind competitively to novel-m0007-5p via 'sponge adsorption' and that interference with LOC110485411 affects hsp90ab1 expression. These results provide potential for anti-stress drug screening in rainbow trout.
虹鳟(Oncorhynchus mykiss)是一种典型的冷水性鱼类。随着全球变暖及极端高温的出现,夏季高温是虹鳟养殖面临的最大威胁。虹鳟会在受到热刺激时启动应激防御机制,而非编码 RNA(microRNAs [miRNAs]、长非编码 RNA)介导的靶基因(mRNA)的竞争内源性 RNA(ceRNA)调控可能是应对热刺激和增强适应的主要策略。
我们筛选了影响虹鳟热应激的 LOC110485411-novel-m0007-5p-hsp90ab1 ceRNA 关系对,并基于初步高通量测序分析结果验证了它们的靶向关系和功能。将外源性 novel-m0007-5p 模拟物和抑制剂转染到原代虹鳟肝细胞中,可有效结合并抑制靶基因 hsp90ab1 和 LOC110485411,而对肝细胞活力、增殖和凋亡无明显影响。 novel-m0007-5p 过表达在热应激下对 hsp90ab1 和 LOC110485411 的抑制作用是高效的。同样,小干扰 RNA(siRNA)通过沉默 LOC110485411 表达,高效地影响 hsp90ab1 mRNA 的表达。
总之,我们发现虹鳟中的 LOC110485411 和 hsp90ab1 可以通过“海绵吸附”的方式与 novel-m0007-5p 竞争结合,干扰 LOC110485411 会影响 hsp90ab1 的表达。这些结果为虹鳟抗应激药物的筛选提供了潜力。