An in situ hybridzation-based screen for heterogeneously expressed genes in mouse ES cells
An in situ hybridzation-based screen for heterogeneously expressed genes in mouse ES cells
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DOI：
10.1016/j.gep.2007.10.009
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发表时间：
2008-02-01
影响因子：
1.2
通讯作者：
Ko, Minoru S. H.
中科院分区：
文献类型：
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作者：
Carter, Mark G.;Stagg, Carole A.;Ko, Minoru S. H.
我们以前报道，Zscan 4在小鼠胚胎干细胞（ES）中表现出异质性表达模式。为了鉴定具有相似表达模式的基因，我们对ES细胞培养物进行了244个基因的高通量原位杂交检测。大多数基因参与转录调控，并选择使用基于微阵列的比较基因表达谱在ES和胚胎癌（EC）细胞与分化的细胞类型。Pou 5 f1（Oct 4，Oct 3/4）和Krt 8（EndoA）用作对照。在ES细胞集落上检测到147个基因（60%）的杂交信号。其中大多数（136个基因）在ES细胞集落中表现出相对均一的表达。然而，我们发现两个基因明确地显示Zscan 4样斑点表达模式（斑点集落模式; Whsc 2和Rhox 9）。我们还发现9个基因表现出相对异质性的表达模式（嵌合体集落模式：Zfp 42/Rex 1、Rest、Atf 4、Pa 2g 4、E2 f2、Nanog、Dppa 31 Pgc 71 Ste 11 a、Esrrb和Fscn 1）。在这些基因中，Zfp 42/Rex 1在通过CytoSpin制备的单个ES细胞中表现出明确的异质性表达。这些结果表明，在ES细胞培养物中存在不同类型或状态的细胞，否则被认为是未分化的和同质的，这表明ES细胞培养物中以前未被认识到的复杂性。由爱思唯尔公司出版
We previously reported that Zscan4 showed heterogeneous expression patterns in mouse embryonic stem (ES) cells. To identify genes that show similar expression patterns, we carried out high-throughput in situ hybridization assays on ES cell cultures for 244 genes. Most of the genes are involved in transcriptional regulation, and were selected using microarray-based comparisons of gene expression profiles in ES and embryonal carcinoma (EC) cells versus differentiated cell types. Pou5f1 (Oct4, Oct3/4) and Krt8 (EndoA) were used as controls. Hybridization signals were detected on ES cell colonies for 147 genes (60%). The majority (136 genes) of them showed relatively homogeneous expression in ES cell colonies. However, we found that two genes unequivocally showed Zscan4-like spotted expression pattern (spot-in-colony pattern; Whsc2 and Rhox9). We also found that nine genes showed relatively heterogeneous expression pattern (mosaicin-colony pattern: Zfp42/Rex1, Rest, Atf4, Pa2g4, E2f2, Nanog, Dppa31Pgc71Ste11a, Esrrb, and Fscn1). Among these genes, Zfp42/Rex1 showed unequivocally heterogeneous expression in individual ES cells prepared by the CytoSpin. These results show the presence of different types or states of cells within ES cell cultures otherwise thought to be undifferentiated and homogeneous, suggesting a previously unappreciated complexity in ES cell cultures. Published by Elsevier B.V.