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在青鳉(Oryzias latipes)的神经发育过程中,miR-124参与了对多嘧啶序列结合蛋白1(PTBP1)的转录后调控。

MiR-124 is involved in post-transcriptional regulation of Polypyrimidine tract binding protein 1 (PTBP1) during neural development in the medaka, Oryzias latipes.

作者信息

Kato Yumiko, Kusakabe Rie, Inoue Kunio, Tochinai Shin

机构信息

1 Department of Natural History Sciences, Graduate School of Science, Hokkaido University, Kita 10 Nishi 8, Kita-ku, Sapporo, Hokkaido 060-0810, Japan.

出版信息

Zoolog Sci. 2013 Nov;30(11):891-900. doi: 10.2108/zsj.30.891.

Abstract

MicroRNAs (miRNAs) comprise a group of small noncoding RNA molecules thought to have contributed to the evolution of vertebrate brain homogeneity and diversity. The miRNA miR-124 is well conserved between invertebrates and vertebrates and is expressed abundantly in the central nervous system (CNS). We identified miR-124 in the medaka, Oryzias latipes, and investigated its role in neural development. The five candidate genes for medaka precursor miR-124 are unlinked on four different chromosomes and differ in nucleotide length. Their sequences suggest that they can generate functional miRNAs through conventional miRNA biogenesis by folding into stem-loop structures. Whole-mount in situ hybridization and northern blotting revealed that mature miR-124 is specifically expressed in the CNS and the eyes starting at two days post-fertilization. We also examined the sequences and expression of medaka Polypyrimidine tract binding protein 1 (Ptbp1), a possible direct target of miR-124. The 3'UTR of medaka Ptbp1 contains predicted binding motifs (target sites) for miR-124. A GFP reporter assay for the target sites or the entire 3'UTR showed that exogenous miR-124 silences PTBP1 expression in vivo. Our study suggests that medaka miR-124 is involved in post-transcriptional regulation of target genes in neural development and that medaka miR-124 homologs may have spatiotemporal roles different from those in other vertebrates.

摘要

微小RNA(miRNA)是一类小的非编码RNA分子,被认为对脊椎动物大脑的同质性和多样性的进化有贡献。miRNA miR-124在无脊椎动物和脊椎动物之间高度保守,并且在中枢神经系统(CNS)中大量表达。我们在青鳉(Oryzias latipes)中鉴定出miR-124,并研究了其在神经发育中的作用。青鳉前体miR-124的五个候选基因在四条不同的染色体上不连锁,并且核苷酸长度不同。它们的序列表明,它们可以通过折叠成茎环结构,通过传统的miRNA生物合成产生功能性miRNA。全胚胎原位杂交和Northern印迹显示,成熟的miR-124在受精后两天开始在中枢神经系统和眼睛中特异性表达。我们还检查了青鳉多嘧啶序列结合蛋白1(Ptbp1)的序列和表达,Ptbp1可能是miR-124的直接靶标。青鳉Ptbp1的3'UTR包含预测的miR-124结合基序(靶位点)。针对靶位点或整个3'UTR的GFP报告基因检测表明,外源性miR-124在体内使PTBP1表达沉默。我们的研究表明,青鳉miR-124参与神经发育中靶基因的转录后调控,并且青鳉miR-124同源物可能具有与其他脊椎动物不同的时空作用。

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