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海胆(Hemicentrotus pulcherrimus)的正常发育需要Dicer。

Dicer is required for the normal development of sea urchin, Hemicentrotus pulcherrimus.

作者信息

Okamitsu Yuka, Yamamoto Takashi, Fujii Takayoshi, Ochiai Hiroshi, Sakamoto Naoaki

机构信息

Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University, 1-3-1 Kagamiyama, Higashi-Hiroshima 739-8526, Japan.

出版信息

Zoolog Sci. 2010 Jun;27(6):477-86. doi: 10.2108/zsj.27.477.

DOI:10.2108/zsj.27.477
PMID:20528154
Abstract

MicroRNAs are single-stranded RNA molecules with a length of 19-25 nucleotides, which play roles in various biological phenomena, including development, differentiation, apoptosis, by regulating target gene expression. Although the presence of microRNA molecules in sea urchin and the expression of genes involved in microRNA biogenesis during sea urchin development have been reported recently, the function of microRNA in sea urchin development remains to be elucidated. In this study, to understand the function of microRNA in the early development of sea urchin, we focused on Dicer, an essential enzyme for biosynthesis of mature microRNA. We determined the nucleotide sequence of cDNA for a Dicer homolog in the sea urchin, Hemicentrotus pulcherrimus, HpDcr, and found that functional domains of Dicer proteins are conserved in HpDcr. Analyses of its pattern of expression showed that HpDcr mRNA is expressed in embryos at all developmental stages analyzed, and seems to distribute asymmetrically at the morula and later stages. Knockdown of HpDcr resulted in anomalous morphogenesis, such as impairment of gastrulation and skeletogenesis at the mesenchyme blastula stage and later stages, and alteration of mRNA levels of cell type-specific genes. Thus, HpDcr plays important roles in morphogenesis in sea urchin embryos, suggesting that miRNA could be involved in the early development of sea urchin by regulating target gene expression.

摘要

微小RNA是长度为19 - 25个核苷酸的单链RNA分子,通过调控靶基因表达在包括发育、分化、凋亡等多种生物学现象中发挥作用。尽管最近有报道称在海胆中存在微小RNA分子以及在海胆发育过程中参与微小RNA生物合成的基因的表达情况,但微小RNA在海胆发育中的功能仍有待阐明。在本研究中,为了解微小RNA在海胆早期发育中的功能,我们聚焦于Dicer,这是成熟微小RNA生物合成所必需的一种酶。我们测定了海胆(光棘球海胆)中Dicer同源物HpDcr的cDNA核苷酸序列,发现Dicer蛋白的功能域在HpDcr中是保守的。对其表达模式的分析表明,HpDcr mRNA在所分析的所有发育阶段的胚胎中均有表达,并且在桑椹胚及之后的阶段似乎呈不对称分布。敲低HpDcr会导致形态发生异常,如在间充质囊胚期及之后阶段原肠胚形成和骨骼形成受损,以及细胞类型特异性基因的mRNA水平改变。因此,HpDcr在海胆胚胎的形态发生中发挥重要作用,这表明微小RNA可能通过调控靶基因表达参与海胆的早期发育。

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