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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。
The aim of this study was to investigate the expression of miR-10a / b in the neural tube of zebrafish during embryonic development and its effects on neuronal development. The expression of miR-10a / b in the neural tube of zebrafish embryos was analyzed by whole-mount in situ hybridization with zebrafish embryo and Taq Man PCR. The miR-10a / b downregulated zebrafish model was established by morpholino (Mo) modified antisense oligonucleotide knockdown technique to study the phenotype of neuronal dysplasia after miR-10a / b downregulation and to identify and identify miR-10 regulates downstream targets of neuronal development. The results showed that miR-10a and miR-10b were highly expressed in the zebrafish neural tube at 24 h post-fertilization (24 hpf) and 48 hpf, while miR-10a / b-Mo downregulated the expression of miR- , The number of neurons in dorsal canal significantly decreased; downregulation of Mib1 (E3 ubiquitin protein ligase 1) could rescue the abnormal phenotype of neurons induced by miR-10 downregulation; Mib1 expression in embryonic neural tube after miR-10a / b downregulation Significantly increased. The above results indicate that miR-10a / b affects the development of zebrafish neurons by inhibiting the expression of Mibl.