Key Laboratory of Agricultural Animal Genetics, Breeding, and Reproduction of Ministry of Education, College of Animal Science and Technology, Huazhong Agricultural University, Wuhan 430070, Hubei, China.
Laboratory Animal Centre, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, Hubei, China.
Biomed Res Int. 2020 Jan 30;2020:4045204. doi: 10.1155/2020/4045204. eCollection 2020.
Alternative splicing (AS) plays a significant role in regulating gene expression at the transcriptional level in eukaryotes. Flexibility and diversity of transcriptome and proteome can be significantly increased through alternative splicing of genes. In the present study, transcriptome data of peripheral immune organs including spleen and inguinal lymph nodes (ILN) were used to identify AS difference between PRRSV-resistant Tongcheng (TC) pigs and PRRSV-susceptible Large White (LW) pigs artificially infected with porcine reproductive and respiratory syndrome virus (PRRSV) . The results showed that PRRSV infection induced global alternative splicing events (ASEs) with different modes. Among them, 373 genes and 595 genes in the spleen and ILN of TC pigs, while 458 genes and 560 genes in the spleen and ILN of LW pigs had significantly differential ASEs. Alternative splicing was subject to tissue-specific and lineage-specific regulation in response to PRRSV infection. Enriched GO terms and pathways showed that genes with differential ASEs played important roles in transcriptional regulation, immune response, metabolism, and apoptosis. Furthermore, a splicing factor associated with apoptosis, , was significantly upregulated in LW pigs. Functional analysis on apoptosis associated genes was validated by RT-PCR and DNA sequencing. These findings revealed different response to PRRSV between PRRSV-resistant TC pigs and PRRSV-susceptible LW pigs at the level of alternative splicing, suggesting the potential relationship between AS and disease resistance to PRRSV.
可变剪接(AS)在真核生物的转录水平上对基因表达的调控起着重要作用。通过基因的可变剪接,可以显著增加转录组和蛋白质组的灵活性和多样性。本研究利用外周免疫器官(包括脾脏和腹股沟淋巴结(ILN))的转录组数据,鉴定了人工感染猪繁殖与呼吸综合征病毒(PRRSV)后 PRRSV 抗性通城(TC)猪和 PRRSV 易感大白猪(LW)之间的 AS 差异。结果表明,PRRSV 感染诱导了不同模式的全基因组可变剪接事件(ASEs)。其中,TC 猪脾脏和 ILN 中有 373 个基因和 595 个基因,LW 猪脾脏和 ILN 中有 458 个基因和 560 个基因发生了显著差异的 ASEs。可变剪接在受到 PRRSV 感染时受到组织特异性和谱系特异性的调节。富集的 GO 术语和途径表明,具有差异 ASEs 的基因在转录调控、免疫反应、代谢和凋亡中发挥重要作用。此外,LW 猪中与凋亡相关的剪接因子 显著上调。通过 RT-PCR 和 DNA 测序对凋亡相关基因进行了功能分析验证。这些发现揭示了 PRRSV 抗性 TC 猪和 PRRSV 易感 LW 猪在可变剪接水平上对 PRRSV 的不同反应,提示 AS 与 PRRSV 抗性之间可能存在关联。