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一项功能基因组学筛选鉴定出一种输入蛋白-α 同源物是涡虫再生过程中干细胞功能和组织模式形成的调节因子。

A functional genomics screen identifies an Importin-α homolog as a regulator of stem cell function and tissue patterning during planarian regeneration.

作者信息

Hubert Amy, Henderson Jordana M, Cowles Martis W, Ross Kelly G, Hagen Matthew, Anderson Christa, Szeterlak Claudia J, Zayas Ricardo M

机构信息

Department of Biology, San Diego State University, San Diego, CA, 92182-4614, USA.

Department of Biological Sciences, Southern Illinois University Edwardsville, Edwardsville, IL, 62026-0001, USA.

出版信息

BMC Genomics. 2015 Oct 12;16:769. doi: 10.1186/s12864-015-1979-1.

Abstract

BACKGROUND

Planarians are renowned for their regenerative capacity and are an attractive model for the study of adult stem cells and tissue regeneration. In an effort to better understand the molecular mechanisms underlying planarian regeneration, we performed a functional genomics screen aimed at identifying genes involved in this process in Schmidtea mediterranea.

METHODS

We used microarrays to detect changes in gene expression in regenerating and non-regenerating tissues in planarians regenerating one side of the head and followed this with high-throughput screening by in situ hybridization and RNAi to characterize the expression patterns and function of the differentially expressed genes.

RESULTS

Along with five previously characterized genes (Smed-cycD, Smed-morf41/mrg-1, Smed-pdss2/dlp1, Smed-slbp, and Smed-tph), we identified 20 additional genes necessary for stem cell maintenance (Smed-sart3, Smed-smarcc-1, Smed-espl1, Smed-rrm2b-1, Smed-rrm2b-2, Smed-dkc1, Smed-emg1, Smed-lig1, Smed-prim2, Smed-mcm7, and a novel sequence) or general regenerative capability (Smed-rbap46/48-2, Smed-mcm2, Smed-ptbp1, and Smed-fen-1) or that caused tissue-specific defects upon knockdown (Smed-ddc, Smed-gas8, Smed-pgbd4, and Smed-b9d2). We also found that a homolog of the nuclear transport factor Importin-α plays a role in stem cell function and tissue patterning, suggesting that controlled nuclear import of proteins is important for regeneration.

CONCLUSIONS

Through this work, we described the roles of several previously uncharacterized genes in planarian regeneration and implicated nuclear import in this process. We have additionally created an online database to house our in situ and RNAi data to make it accessible to the planarian research community.

摘要

背景

涡虫以其再生能力而闻名,是研究成体干细胞和组织再生的理想模型。为了更好地理解涡虫再生的分子机制,我们进行了一项功能基因组学筛选,旨在鉴定参与地中海涡虫这一过程的基因。

方法

我们使用微阵列检测头部一侧再生和未再生组织中基因表达的变化,随后通过原位杂交和RNA干扰进行高通量筛选,以表征差异表达基因的表达模式和功能。

结果

除了五个先前已表征的基因(Smed-cycD、Smed-morf41/mrg-1、Smed-pdss2/dlp1、Smed-slbp和Smed-tph)外,我们还鉴定出另外20个对干细胞维持(Smed-sart3、Smed-smarcc-1、Smed-espl1、Smed-rrm2b-1、Smed-rrm2b-2、Smed-dkc1、Smed-emg1、Smed-lig1、Smed-prim2、Smed-mcm7和一个新序列)、一般再生能力(Smed-rbap46/48-2、Smed-mcm2、Smed-ptbp1和Smed-fen-1)必不可少的基因,或在敲低后导致组织特异性缺陷的基因(Smed-ddc、Smed-gas8、Smed-pgbd4和Smed-b9d2)。我们还发现核转运因子输入蛋白-α的一个同源物在干细胞功能和组织模式形成中起作用,这表明蛋白质的可控核输入对再生很重要。

结论

通过这项工作,我们描述了几个先前未表征的基因在涡虫再生中的作用,并表明核输入参与了这一过程。我们还创建了一个在线数据库来存储我们的原位和RNA干扰数据,以便涡虫研究界能够访问。

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