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Cell-Specific mRNA Profiling of the Caenorhabditis elegans Somatic Gonadal Precursor Cells Identifies Suites of Sex-Biased and Gonad-Enriched Transcripts

机译:秀丽隐杆线虫体细胞性腺前体细胞的细胞特异mRNA谱鉴定了性偏爱和性腺富集转录本的套件。

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摘要

The Caenorhabditis elegans somatic gonad differs greatly between the two sexes in its pattern of cell divisions, migration, and differentiation. Despite decades of study, the genetic pathways directing early gonadal development and establishing sexual dimorphism in the gonad remain largely unknown. To help define the genetic networks that regulate gonadal development, we employed cell-specific RNA-seq. We identified transcripts present in the somatic gonadal precursor cells and their daughter cells of each sex at the onset of sexual differentiation. We identified several hundred gonad-enriched transcripts, including the majority of known regulators of early gonadal development, and transgenic reporter analysis confirmed the effectiveness of this approach. Before the division of the somatic gonad precursors, few sex-biased gonadal transcripts were detectable; less than 6 hr later, after their division, we identified more than 250 sex-biased transcripts, of which about a third were enriched in the somatic gonad compared to the whole animal. This indicates that a robust sex-biased developmental program, some of it gonad-specific, initiates in the somatic gonadal precursor cells around the time of their first division. About 10% of male-biased transcripts had orthologs with male-biased expression in the early mouse gonad, suggesting possible conservation of gonad sex differentiation. Cell-specific analysis also identified approximately 70 previously unannotated mRNA isoforms that are enriched in the somatic gonad. Our data illustrate the power of cell-specific transcriptome analysis and suggest that early sex differentiation in the gonad is controlled by a relatively small suite of differentially expressed genes, even after dimorphism has become apparent.
机译:秀丽隐杆线虫的体细胞性腺在细胞分裂,迁移和分化的模式上有很大的不同。尽管进行了数十年的研究,但是指导性腺早期发育和在性腺中建立性二态性的遗传途径仍然未知。为了帮助定义调节性腺发育的遗传网络,我们采用了细胞特异性RNA-seq。我们鉴定了在性分化开始时每种性别的体细胞性腺前体细胞及其子代细胞中存在的转录本。我们鉴定了数百种富含性腺的转录本,包括大多数已知的早期性腺发育调控因子,而转基因记者分析证实了这种方法的有效性。在体细胞性腺前体分裂之前,几乎没有检测到性别偏向的性腺转录本。不到6小时后,在将它们分开后,我们鉴定出250多个性别偏向性转录本,与整个动物相比,其中约有1/3富含体细胞性腺。这表明一个强大的性别偏见的发育程序(其中有些是性腺特异的)在第一次分裂时就在体细胞性腺前体细胞中启动。大约10%的男性偏向性转录本在早期的小鼠性腺中具有直系同源表达且男性偏向的表达,这表明可能保留了性腺的性别分化。细胞特异性分析还确定了大约70种先前未注释的mRNA异构体,这些异构体富含体细胞的性腺。我们的数据说明了细胞特异性转录组分析的能力,并表明性腺的早期性别分化受相对较小的一组差异表达基因控制,即使在二态性变得明显之后也是如此。

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