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目的:研究n elavl1/HuR对斑马鱼肠神经系统早期发育的影响。n 方法:应用免疫组化方法检测elavl1、HuC/HuD在野生型斑马鱼肠道的表达。挑选25个斑马鱼胚胎,应用Western Blot法检测elavl1在受精后24~96 h野生型斑马鱼中的蛋白表达水平。在野生型斑马鱼胚胎处于1细胞期时分别显微注射4 ng、8 ng n elavl1a morpholino和8 ng control morpholino,挑选25个受精后24 h的胚胎采用Western Blot法检测elavl1的蛋白表达,免疫组化检测受精后72 h、96 h的胚胎斑马鱼肠道神经元的分布情况。n 结果:elavl1在斑马鱼全身都有表达,而HuC/HuD在第四天斑马鱼肠道部位出现表达;在受精后24~96 h时期,elavl1在斑马鱼中的蛋白表达量随斑马鱼生长而增加,且受精后96 h时elavl1蛋白量明显高于受精后24 h、48 h、72 h,差异有统计学意义(n P<0.05);分别注射4 ng、8 ngn elavl1a Mo和8 ng Con Mo后,发现注射8 ng Con Mo组elavl1蛋白含量高于其他两组,且差异有统计学意义(n P<0.05);敲降elavl1a后免疫组化显示在受精后72 h时三组都不能在肠道末端明显观察到神经元,受精后96 h时可以观察到n elavl1a Mo 8 ng组和n elavl1a Mo 4 ng组较Con Mo 8 ng组斑马鱼肠神经元数量有所减少,每组随机抽取5条斑马鱼统计肠道末端到前面四个体节长度的肠道神经元数量,发现n elavl1a Mo 8 ng组和n elavl1a Mo 4 ng组较Con Mo 8 ng组斑马鱼肠神经元数量差异有统计学意义(n P<0.05)。n 结论:elavl1a是斑马鱼肠神经发育过程中重要基因,敲降elavl1a会减少肠神经元细胞数量。n “,”Objective:To explore the effect of n elavl1/HuR on early development of zebrafish enteric nervous system.n Methods:Immunohistochemistry was employed for detecting the expressions of elavl1 and HuC/HuD in intestines of wild-type zebrafish. Twenty-five zebrafish embryos were selected and Western blot was utilized for detecting the protein level of elavl1 in wild-type embryos during 24hpf-96hpf. 4/8 ng n elavl1 morpholino or 8 ng control morpholino were injected into wild-type embryos at unicellular stage. Twenty-five embryos were selected for detecting the expression level of elavl1a in 24hpf-96hpf embryos by Western blot and enteric neurons in 72/96hpf embryos by immunohistochemistry.n Results:It was found that elavl1 was expressed ubiquitously while HuC/HuD only in intestinal tract in 96 hpf. elavl1 protein gradually increased with embryonic development at 24hpf-96hpf and elavl1 protein at 96hpf was significantly higher than that at 24/48/72hpf (n P<0.05). Moreover, after an injection of 4/8 ngn elavl1a morpholino and 8ng Con Mo respectively, the protein level of elavl1 was statistically higher in control group than that of n elavl1a morpholino injected groups (n P<0.05). After knockdown of elavl1, immunohistochemistry indicated that neurons disappeared at the end of intestine at 72hpf. At 96hpf, the count of intestinal neurons declined inn elavl1a morpholino injected groups as compared to control group. Five zebrafishes were randomly selected from each group for counting the number of intestinal neurons from the end of intestinal tract to four front segments. There was a significant reduction of intestinal neurons in n elavl1a morpholino injected groups as compared to control group (n P<0.05).n Conclusions:Thus n elavl1a plays a critical role in the development of zebrafish enteric nervous system and its knockdown reduces the count of enteric neurons.n