For intensive aquaculture in freshwater ponds, microcystin (MC-LR) generated from cyanobacterial blooms is one of the bottlenecks for the healthy and sustainable development of shrimp aquaculture industry. In this study, we measured the MC-LR content in the hepatopancreas and muscles of Litopenaeus vannamei stressed by MC-LR, and analyzed protein expression in the hepatopancreas using DIA high-throughput proteomics technology. The results showed that MC-LR content in the hepatopancreas and muscles reached the highest at 1 h after MC-LR injection, which was (6.12±0.45) μg·kg-1 and (5.00±0.19) μg·kg-1, respectively. Then, it decreased gra-dually, with that in the hepatopancreas being significantly higher than in muscles. We identified 820 differential expressed proteins, including 586 up-regulated and 234 down-regulated ones. Results of bioinformatics analysis showed that MC-LR stress significantly affected immune-related pathways such as lysosome, RIG-Ⅰ receptor signals and interleukin-2. It also altered energy metabolisms including citrate cycle, metabolism of starch and sucrose, and interconversion of pentose and glucoronate, which in turn led to the disorder of carbohydrate metabolism. In addition, MC-LR significantly upregulated 19 cytoskeleton-related blood shadow proteins and damaged the hepatopancreas cytoskeleton. It was concluded that MC-LR mainly affected the physiological processes associated with immunity, energy metabolism, and cytoskeleton in the hepatopancreas of L. vannamei.
在淡水池塘集约化养殖中,蓝藻水华爆发产生的微囊藻毒素(MC-LR)是制约南美白对虾养殖业健康可持续发展的瓶颈之一。本研究在分析MC-LR胁迫条件下凡纳滨对虾肝胰腺和肌肉中MC-LR含量的基础上,利用DIA高通量蛋白质组学技术探究对虾肝胰腺蛋白表达情况。结果表明: 注射MC-LR 1 h后,肝胰腺和肌肉中毒素含量达到最高,分别为(6.12±0.45)和(5.00±0.19) μg·kg-1,随后逐渐降低,但肝胰腺中毒素含量始终显著高于肌肉;共鉴定820个差异蛋白,其中上调蛋白586个,下调蛋白234个。通过生物信息学分析发现,MC-LR胁迫显著影响溶酶体、RIG-Ⅰ样受体信号和白细胞介素-2等免疫通路;在能量代谢方面主要影响柠檬酸循环、淀粉与蔗糖代谢和戊糖与葡萄糖醛酸转换等,导致碳水化合物代谢紊乱;引起19个与细胞骨架相关的血影蛋白显著上调,破坏对虾肝胰腺细胞骨架。表明MC-LR主要对凡纳滨对虾肝胰腺的免疫、能量代谢和细胞骨架产生影响。.
Keywords:
DIA.;