microRNA-10a/b通过抑制Mib1调节斑马鱼神经元的发育  

microRNA-10a/b Regulate Neuron Development by Repressing Mib1 in Zebrafish

作　　者：吕峰[1,2] 石运伟[3] 王新[3] 刘东[3] 严兴洪[1] 

机构地区：[1]上海海洋大学水产与生命学院,上海201306 [2]南通科技职业学院,南通226007 [3]南通大学江苏省神经再生重点实验室,南通226001

出　　处：《中国细胞生物学学报》2017年第2期191-196,共6页Chinese Journal of Cell Biology

基　　金：南通大学自然科学基金(批准号:12z053);南通市科技项目2015N应用基础研究–社会民生创新–现代农业(批准号:MS12015072)资助的课题~~

摘　　要：该文研究了microRNA-10(miR-10)家族miR-10a/b在斑马鱼胚胎发育时期神经管的表达及其对神经元发育的影响。通过斑马鱼胚胎整体原位杂交及Taq Man PCR技术分析研究miR-10a/b在斑马鱼胚胎期神经管的表达情况。利用吗啡啉(morpholino,Mo)修饰的反义寡核苷酸敲低技术建立miR-10a/b下调的斑马鱼模型,研究miR-10a/b下调后神经元发育异常的表型,并分析鉴定miR-10调控神经元发育的下游靶点。结果发现,受精后24 h(24 hours post fertilization,24 hpf)和48 hpf,miR-10a和miR-10b在斑马鱼神经管中高表达;miR-10a/b-Mo下调miR-10a/b的表达后,背神经管中神经元数量明显变少;下调Mib1(mindbomb E3 ubiquitin protein ligase 1)能挽救miR-10下调引起的神经元表型异常;miR-10a/b下调后胚胎神经管中Mib1表达显著上调。上述结果表明,miR-10a/b通过抑制Mib1的表达来影响斑马鱼神经元的发育。To study the role of miR-10a/b in neuronal development of zebrafish, firstly we investigated the expression of miR-10a/b in embryonic neural tube by whole-mount in situ hybridization and Taq Man PCR. Then we established zebrafish model with miR-10a/b knockdown using morpholino（Mo） oligonucleotide antisenses. Based on this model, we monitored the abnormal phenotype of neuron in neural tube, and further analyzed whether down-regulated mindbomb E3 ubiquitin protein ligase 1（Mib1） rescue abnormal phenotype caused by miR-10a-Mo injection. We found miR-10 a and miR-10 b were highly expressed in the zebrafish neural tube at 24 hours post fertilization（hpf） and 48 hpf. Neurons in dorsal neural tube were obviously reduced by miR-10a/b knockdown. In addition, down-regulated Mib1 could partially rescue neuron phenotype defects induced by miR-10 down-regulation and m RNA level of Mib1 in miR-10-Mo injected embryos was apparently increased. These results indicated that miR-10a/b affected the development of neurons in zebrafish by repressing the Mib1.

关 键 词：miR-10a MIR-10B 神经元 发育 Mib1 斑马鱼 

分 类 号：R-33[医药卫生]