Hou Fujun, He Shulin, Liu Yongjie, Zhu Xiaowen, Sun Chengbo, Liu Xiaolin
College of Animal Science and Technology, Northwest A&F University, Shaanxi Key Laboratory of Molecular Biology for Agriculture, Yangling 712100, China.
Fisheries College, Guangdong Ocean University, Guangdong 524088, China.
Dev Comp Immunol. 2014 Jun;44(2):255-60. doi: 10.1016/j.dci.2014.01.004. Epub 2014 Jan 14.
NF-κB dependent antimicrobial peptides (AMPs) are of critical importance in protecting insects or mammals from microorganisms infection. However, we still do not make clear signaling pathways in regulating AMPs expression in shrimps. In this study, RNAi approach was used to study differences between Toll signaling pathway and immune deficiency signaling pathway in regulating the transcription of NF-κB dependent AMPs post bacteria challenge. Results showed that the transcription level of anti-lipopolysaccharide factor was highly suppressed in Litopenaeus vannamei immune deficiency (LvIMD) silenced shrimps by gene specific dsRNA compared to Litopenaeus vannamei Toll (LvToll) silenced shrimps with or without Vibrio anguillarum and Micrococcus lysodeikticus challenge. Conversely the transcription level of penaeidin3a was significantly suppressed in LvToll silenced shrimps compared to LvIMD silenced shrimps. However, no obvious difference was found in regulating the transcription of CrustinP. Meanwhile, we found that silencing LvToll both down regulated the transcription of Dorsal and Relish while silencing LvIMD only down regulated the transcription of Relish. At last, shrimp survival experiment showed that post V. anguillarum challenge high mortality was found both in LvToll and LvIMD silenced groups while post M. lysodeikticus challenge we saw high mortality only in LvToll silenced group. Hence, we conclude that shrimp L. vannamei Toll pathway and IMD pathway might be different in regulating the transcription of NF-κB dependent AMPs and responding to bacteria challenge but not independent of each other.
核因子κB依赖性抗菌肽(AMPs)在保护昆虫或哺乳动物免受微生物感染方面至关重要。然而,我们仍不清楚对虾中调控AMPs表达的信号通路。在本研究中,采用RNA干扰方法研究Toll信号通路和免疫缺陷信号通路在细菌攻击后调控核因子κB依赖性AMPs转录方面的差异。结果表明,与经鳗弧菌和溶壁微球菌攻击或未攻击的凡纳滨对虾Toll(LvToll)基因沉默对虾相比,凡纳滨对虾免疫缺陷(LvIMD)基因沉默对虾中抗脂多糖因子的转录水平受到基因特异性双链RNA的高度抑制。相反,与LvIMD基因沉默对虾相比,LvToll基因沉默对虾中对虾素3a的转录水平受到显著抑制。然而,在调控CrustinP的转录方面未发现明显差异。同时,我们发现沉默LvToll会下调背侧蛋白和Relish的转录,而沉默LvIMD仅下调Relish的转录。最后,对虾存活实验表明,在鳗弧菌攻击后,LvToll和LvIMD基因沉默组均出现高死亡率,而在溶壁微球菌攻击后,仅在LvToll基因沉默组出现高死亡率。因此,我们得出结论,凡纳滨对虾Toll通路和IMD通路在调控核因子κB依赖性AMPs转录及应对细菌攻击方面可能存在差异,但并非相互独立。