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出生后发育中小鼠视网膜的全基因组表达分析。

Global gene expression analysis of the developing postnatal mouse retina.

作者信息

Dorrell Michael I, Aguilar Edith, Weber Christoph, Friedlander Martin

机构信息

Departments of Cell Biology, The Scripps Research Institute, La Jolla, California 92037, USA.

出版信息

Invest Ophthalmol Vis Sci. 2004 Mar;45(3):1009-19. doi: 10.1167/iovs.03-0806.

Abstract

PURPOSE

Postnatal mouse retinal development involves glial and neuronal differentiation, vascularization, and the onset of vision. In the current study, the gene expression profiles of thousands of genes in the developing postnatal mouse retina were analyzed and compared in a large-scale, unbiased microarray gene expression analysis.

METHODS

For each of eight different time points during postnatal mouse retinal development, two separate sets of 30 retinas were pooled for RNA isolation, and gene expression was analyzed by hybridization to gene chips in triplicate (Mu74Av2; Affymetrix, Santa Clara, CA). Genes were sorted into clusters based on their expression profiles and intensities. Validation was accomplished by comparing the microarray expression profiles with real-time RT-PCR analysis of selected genes and by comparing selected expression profiles with predicted profiles based on previous studies.

RESULTS

The Mu74Av2 chip contains more than 6000 known genes and 6500 estimated sequence tags (ESTs) from the mouse Unigene database. Of these, 2635 known gene sequences and 2794 ESTs were expressed at least threefold above background levels during retinal development. Expressed genes were clustered based on expression profiles allowing potential functions for specific genes during retinal development to be inferred by comparison to developmental events occurring at each time point. Specific data and potential functions for genes with various profiles are discussed. All data can be viewed online at http://www.scripps.edu/cb/friedlander/gene_expression/.

CONCLUSIONS

Expression analysis of thousands of different genes during normal postnatal mouse retinal development as reported in this study demonstrates that such an approach can be used to correlate gene expression with known functional differentiation, presenting the opportunity to infer functional correlates between gene expression and specific postnatal developmental events.

摘要

目的

出生后小鼠视网膜发育涉及神经胶质细胞和神经元分化、血管生成以及视觉的开始。在本研究中,在大规模、无偏倚的微阵列基因表达分析中,对出生后发育中的小鼠视网膜中数千个基因的基因表达谱进行了分析和比较。

方法

在出生后小鼠视网膜发育的八个不同时间点,每组收集30个视网膜,共两组,用于RNA分离,并通过与基因芯片杂交进行三次重复分析基因表达(Mu74Av2;Affymetrix,加利福尼亚州圣克拉拉)。根据基因的表达谱和强度将其分类为簇。通过将微阵列表达谱与所选基因的实时RT-PCR分析进行比较,以及将所选表达谱与基于先前研究的预测谱进行比较来完成验证。

结果

Mu74Av2芯片包含来自小鼠Unigene数据库的6000多个已知基因和6500个估计序列标签(EST)。其中,2635个已知基因序列和2794个EST在视网膜发育过程中表达水平至少比背景水平高3倍。根据表达谱对表达的基因进行聚类,通过与每个时间点发生的发育事件进行比较,可以推断特定基因在视网膜发育过程中的潜在功能。讨论了具有各种谱的基因的具体数据和潜在功能。所有数据可在http://www.scripps.edu/cb/friedlander/gene_expression/在线查看。

结论

本研究报道的正常出生后小鼠视网膜发育过程中数千个不同基因的表达分析表明,这种方法可用于将基因表达与已知的功能分化相关联,为推断基因表达与特定出生后发育事件之间的功能相关性提供了机会。

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