为探明JNK信号通路在鱼类胚胎发育过程中的作用,通过使用JNK信号特异性抑制剂SP600125对金鱼胚胎进行浸泡处理,采用荧光实时定量PCR和蛋白免疫印迹技术检测JNK1 mRNA和蛋白在胚胎不同发育时期的表达.结果显示:从受精卵发育至原肠胚期后,SP600125能够显著阻滞胚胎的发育时间,同时伴随大脑、眼睛、心脏和体节等组织器官的发育迟缓并出现畸形.qPCR检测结果显示JNK1在实验组和对照组中的表达模式没有明显变化,均表现为"升高-降低-升高-降低"的趋势.Western Blotting 检测结果显示JNK1蛋白的表达在对照组中的表达模式与mRNA的表达水平吻合,而实验组中,胚胎发育至原肠胚后JNK1蛋白持续保持较低水平的增加.研究表明,JNK信号通路在鱼类胚胎发育和组织器官发生及发育过程中发挥了重要作用.%The objective of this study was to explore the correlation of the c-Jun amino-terminal kinase(JNK)signaling pathway in the process of embryonic development of Goldfish(Carassius auratus).SP600125,a JNK-specific inhibitor,gives rise to high mortality in goldfish;the expression of JNK1 was studied and analyzed in different developmental phases of embryo by using Quantitative Real -time PCR and Western Blotting.It was found that SP600125 could significantly block the embryo development time after gastrula stage. SP600125 is also responsible for the severe abnormality of organs,especially the brain,eyes,heart and skeletal system.The qPCR a-nalysis revealed that JNK1 appeared as "high-low-high-low"expression pattern during different stages of embryonic development in ex -perimental group and control group.Western Blotting analysis showed that expression tendency of JNK 1 protein was in line with that of JNK1 mRNA in the control group.However,in the experimental group,JNK1 protein maintained a low level increase after gastrula sta-ges compare to the control group.Together, these results suggested that JNK signaling pathway plays an important role in embryonic development and organogenesis of fish.
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