High-throughput screen for genes predominantly expressed in the ICM of mouse blastocysts by whole mount in situ hybridization

被引:69
作者
Yoshikawa, T
Piao, YL
Zhong, JH
Matoba, R
Carter, MG
Wang, YX
Goldberg, I
Ko, MSH [1 ]
机构
[1] NIA, Dev Genom & Aging Sect, Genet Lab, NIH, Baltimore, MD 21224 USA
[2] NIA, Image Informat & Computat Biol Unit, Genet Lab, NIH, Baltimore, MD 21224 USA
关键词
preimplantation embryo; whole mount in situ hybridization; high-throughput screen; hybridization chamber; ICM; TE;
D O I
10.1016/j.modgep.2005.06.003
中图分类号
Q [生物科学];
学科分类号
07 ; 0710 ; 09 ;
摘要
Mammalian preimplantation embryos provide an excellent opportunity to study temporal and spatial gene expression in whole mount in situ hybridization (WISH). However, large-scale studies are made difficult by the size of the embryos (similar to 60 mu m diameter) and their fragility. We have developed a chamber system that allows parallel processing of embryos without the aid of a microscope. We first selected 91 candidate genes that were transcription factors highly expressed in blastocysts, and more highly expressed in embryonic (ES) than in trophoblast (TS) stem cells. We then used the WISH to identify 48 genes expressed predominantly in the inner cell mass (ICM) and to follow several of these genes in all seven preimplantation stages. The ICM-predominant expressions of these genes suggest their involvement in the pluripotency of embryonic cells. This system provides a useful tool to a systematic genome-scale analysis of preimplantation embryos. Published by Elsevier B.V.
引用
收藏
页码:213 / 224
页数:12
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