Hipolito Sheryll Grospe, Shitara Aiko, Kondo Hidehiro, Hirono Ikuo
Graduate School of Marine Science and Technology, Tokyo University of Marine Science and Technology, Konan 4-5-7, Minato-ku, Tokyo 108-8477, Japan.
Graduate School of Marine Science and Technology, Tokyo University of Marine Science and Technology, Konan 4-5-7, Minato-ku, Tokyo 108-8477, Japan.
Dev Comp Immunol. 2014 Oct;46(2):461-9. doi: 10.1016/j.dci.2014.06.001. Epub 2014 Jun 11.
Crustins are important AMP that has been identified in crustaceans. In this study, the role of Marsupenaeus japonicus crustin-like peptide (MjCRS) was examined in vivo by RNA interference (RNAi) using double-stranded RNA (dsRNA). Tissue expression analysis revealed that MjCRS transcripts are expressed in different tissues tested with the highest expression observed in hemocytes. Treatment with double-stranded RNA specific to MjCRS led to a significant reduction of MjCRS transcripts within the hemocytes. When MjCRS was silenced and subsequently infected with Vibrio penaeicida final mortality was significantly higher compared with PBS and dsGFP treated groups. On the other hand, final mortalities of MjCRS silenced and PBS injected groups were not significantly different after infection with white spot virus, however, both are significantly higher compared with dsGFP treated group. V. penaeicida infection significantly decreased MjCRS expression at 3, 6, 12 and 24h followed by significant increase at 48 h post-infection. On the contrary, white spot infection significantly increased MjCRS expression at 6 and 12h and decreased at 48 h post-infection. dsRNA treatment alone decreased total hemocyte counts (THCs) and subsequent V. penaeicida or white spot virus infection further decreased THCs. VP28 gene expression was both similarly increased in PBS injected group and MjCRS silenced group at 24 and 48 h-post infection. Results suggest that MjCRS is involved in antibacterial defense and might not have critical function against viral infection.
甲壳素是在甲壳类动物中发现的重要抗菌肽。在本研究中,通过使用双链RNA(dsRNA)的RNA干扰(RNAi)在体内检测了日本对虾类甲壳素样肽(MjCRS)的作用。组织表达分析表明,MjCRS转录本在不同测试组织中均有表达,其中血细胞中的表达量最高。用针对MjCRS的双链RNA处理导致血细胞中MjCRS转录本显著减少。当MjCRS被沉默,随后感染致病性弧菌时,最终死亡率与PBS和dsGFP处理组相比显著更高。另一方面,在感染白斑病毒后,MjCRS沉默组和PBS注射组的最终死亡率没有显著差异,然而,两者均显著高于dsGFP处理组。致病性弧菌感染在感染后3、6、12和24小时显著降低MjCRS表达,但在感染后48小时显著增加。相反,白斑感染在感染后6和12小时显著增加MjCRS表达,在感染后48小时降低。单独的dsRNA处理降低了总血细胞计数(THCs),随后致病性弧菌或白斑病毒感染进一步降低了THCs。在感染后24和48小时,PBS注射组和MjCRS沉默组中的VP28基因表达均同样增加。结果表明,MjCRS参与抗菌防御,可能对病毒感染没有关键作用。