Donghu Experimental Station of Lake Ecosystems, State Key Laboratory for Freshwater Ecology and Biotechnology of China, The Chinese Academy of Sciences, Wuhan, P R China.
Proteomics. 2011 May;11(10):2003-18. doi: 10.1002/pmic.201000442. Epub 2011 Apr 7.
Microcystin-leucine-arginine (MCLR) is the most toxic and the most commonly encountered variant of microcystins (MCs) in aquatic environment, and it has the potential for developmental toxicity. A number of previous studies have described the developing toxicity of MCLR based on conventional toxicological indices. However, the molecular mechanisms by which it expresses its toxicity during the early development remain largely unknown. To further our understanding of mechanisms of action and identify the potential protein biomarkers for MCLR exposure, a proteomic analysis was performed on developing zebrafish embryos exposed to 0.5 mg/L MCLR until 96 hours post-fertilization. 2-DE combined with MS was employed to detect and identify the protein profiles. Results showed that 75 spots from the 0.5 mg/L MCLR condition showed a significant increase or decrease in abundance compared with the control. In total, 40 proteins were identified. These proteins were mainly included in process related to oxidative stress, energetic metabolism, and the cytoskeleton assembly. MCLR exposure also affects the expression of the subunits of protein phosphatases 2A. Furthermore, the proteomic and transcriptional analysis of nine proteins was determined by Western blot and quantitative real-time PCR due to their correlation with the known MCLR toxic mechanisms. The consistent and discrepant results between protein and mRNA levels indicated complicated regulatory mechanisms of gene expression in response to MCLR exposure.
微囊藻毒素亮氨酸精氨酸(MCLR)是水生环境中最具毒性和最常见的微囊藻毒素(MCs)变体,具有潜在的发育毒性。许多先前的研究已经基于传统的毒理学指标描述了 MCLR 的发育毒性。然而,它在早期发育过程中表达毒性的分子机制在很大程度上仍然未知。为了进一步了解作用机制,并确定潜在的 MCLR 暴露的蛋白质生物标志物,对暴露于 0.5mg/L MCLR 的发育中的斑马鱼胚胎进行了蛋白质组学分析,直到受精后 96 小时。采用 2-DE 联合 MS 检测和鉴定蛋白质图谱。结果表明,与对照组相比,0.5mg/L MCLR 条件下的 75 个斑点的丰度显著增加或减少。总共鉴定出 40 种蛋白质。这些蛋白质主要包括与氧化应激、能量代谢和细胞骨架组装相关的过程。MCLR 暴露还会影响蛋白磷酸酶 2A 的亚基表达。此外,由于与已知的 MCLR 毒性机制相关,通过 Western blot 和定量实时 PCR 确定了 9 种蛋白质的蛋白质组学和转录分析。蛋白质和 mRNA 水平之间的一致和不一致结果表明,基因表达对 MCLR 暴露的复杂调控机制。