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秀丽隐杆线虫的胚胎肌肉转录组

The embryonic muscle transcriptome of Caenorhabditis elegans.

作者信息

Fox Rebecca M, Watson Joseph D, Von Stetina Stephen E, McDermott Joan, Brodigan Thomas M, Fukushige Tetsunari, Krause Michael, Miller David M

机构信息

Department of Cell and Developmental Biology, Vanderbilt University, 465 21st Ave. S., Nashville, TN 37232-8240, USA.

出版信息

Genome Biol. 2007;8(9):R188. doi: 10.1186/gb-2007-8-9-r188.

Abstract

BACKGROUND

The force generating mechanism of muscle is evolutionarily ancient; the fundamental structural and functional components of the sarcomere are common to motile animals throughout phylogeny. Recent evidence suggests that the transcription factors that regulate muscle development are also conserved. Thus, a comprehensive description of muscle gene expression in a simple model organism should define a basic muscle transcriptome that is also found in animals with more complex body plans. To this end, we applied microarray profiling of Caenorhabtidis elegans cells (MAPCeL) to muscle cell populations extracted from developing C. elegans embryos.

RESULTS

We used fluorescence-activated cell sorting to isolate myo-3::green fluorescent protein (GFP) positive muscle cells, and their cultured derivatives, from dissociated early C. elegans embryos. Microarray analysis identified 7,070 expressed genes, 1,312 of which are enriched in the myo-3::GFP positive cell population relative to the average embryonic cell. The muscle enriched gene set was validated by comparisons with known muscle markers, independently derived expression data, and GFP reporters in transgenic strains. These results confirm the utility of MAPCeL for cell type specific expression profiling and reveal that 60% of these transcripts have human homologs.

CONCLUSION

This study provides a comprehensive description of gene expression in developing C. elegans embryonic muscle cells. The finding that more than half of these muscle enriched transcripts encode proteins with human homologs suggests that mutant analysis of these genes in C. elegans could reveal evolutionarily conserved models of muscle gene function, with ready application to human muscle pathologies.

摘要

背景

肌肉产生力量的机制在进化上由来已久;肌节的基本结构和功能组件在整个系统发育过程中是所有活动动物所共有的。最近的证据表明,调节肌肉发育的转录因子也是保守的。因此,在一个简单的模式生物中对肌肉基因表达进行全面描述,应该能够定义一个在身体结构更复杂的动物中也能找到的基本肌肉转录组。为此,我们将秀丽隐杆线虫细胞微阵列分析(MAPCeL)应用于从发育中的秀丽隐杆线虫胚胎中提取的肌肉细胞群体。

结果

我们使用荧光激活细胞分选技术从解离的早期秀丽隐杆线虫胚胎中分离出肌球蛋白3::绿色荧光蛋白(GFP)阳性的肌肉细胞及其培养衍生物。微阵列分析鉴定出7070个表达基因,其中1312个相对于平均胚胎细胞在肌球蛋白3::GFP阳性细胞群体中富集。通过与已知肌肉标记物、独立获得的表达数据以及转基因菌株中的GFP报告基因进行比较,验证了肌肉富集基因集。这些结果证实了MAPCeL在细胞类型特异性表达谱分析中的实用性,并揭示这些转录本中有60%具有人类同源物。

结论

本研究全面描述了发育中的秀丽隐杆线虫胚胎肌肉细胞中的基因表达。这些肌肉富集转录本中超过一半编码具有人类同源物的蛋白质,这一发现表明在秀丽隐杆线虫中对这些基因进行突变分析可能揭示肌肉基因功能的进化保守模型,并可直接应用于人类肌肉病理学研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ece4/2375026/4b7fffeab7e5/gb-2007-8-9-r188-1.jpg

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