Studies on Stress Response Related Genes Affected by Acrylamide in Zebrafish (Danio rerio)
XU Yue1,2, LIANG Li-Qun2,*, SUN Bo2, CHANG Yu-Mei2, ZHAO Xue-Fei3
1 National Demonstration Center for Experimental Fisheries Science Education, Shanghai Ocean University, Shanghai 201306, China;
2 Heilongjiang River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Harbin 150070, China;
3 College of Wildlife and Protected Area, Northeast Forestry University, Harbin 150040, China
Abstract：Acrylamide (ACR), one of the critical environmental stressors, is widely used in soil stabilization, water treatment, industrial products, and certain foods, which toxicity was attracting global attention. This study focused on the toxic effects of ACR in non-neural organs such as the intestines. This study used 3-day-old zebrafish (Danio rerio) and laboratory-grown zebrafish intestine cells (ZFI) as experimental materials. Coagulant fluorescence markers and qRT-PCR were used to check some stress response genes, glutathione-disulfide reductase (GSR), X-box binding protein 1 (xbp1u), recombinant aquaporin 4 (aqp4), and calcium/calmodulin-dependent-protein (CaMK2g2) for expression testing. The 4 genes' functions were related to reaction mechanisms such as oxidative stress, endosurgen network stress, permeable pressure stress, and stress signal-transducing. The coagulation point imprinting results showed that the maackia amurensis lectin Ⅱ(MAL-Ⅱ), wheat germ agglutini (WGA), and peanut agglutinin (PNA) tricoagulant bars were at 12.5 and 25.0 mmol/L tolerances than the 0 mmol/L ACR tolerance. The bands showed varying degrees of shallow changes in zebrafish and ZFI in 45~60 kD, where the changes in the coagulant bands of ZFI at a tolerance concentration of 25.0 mmol/L were more pronounced. Besides, at a concentration of 25.0 mmol/L ACR, ACR toxicity caused the most significant damage to the body (18 h) and cells (5 h), and the above 4 genes associated with stress response showed that for ZFI, aqp4 and xbp1u showed very high significant expression (P<0.01), CaMK2g2 and GSR showed significant high expression in ZFI (P<0.05); For zebrafish, the expression of aqp4 increased significantly (P<0.05), xbp1u and GSR showed extremely high significant expression (P<0.01), and CaMK2g2 showed no significant change (P>0.05). The above results showed that ACR toxicity was systemic when it caused damage to cells, and the ACR toxicity mechanism was closely related to the non-biological stress response mechanism. Therefore, this study's significance lied in revealing the fish bodies' stress response mechanism to ACR stress. Multi-level interpretation of its toxicity mechanism would be conducive to protect fish and other aquatic organisms from ACR toxic intrusion to meet the growing aquatic food safety needs of China and the world's population.
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