High-throughput screen for genes predominantly expressed in the ICM of mouse blastocysts by whole mount in situ hybridization.
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 ( approximately 60mum 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.
Yoshikawa
Toshiyuki
T
Developmental Genomics and Aging Section, Laboratory of Genetics, National Institute on Aging, National Institutes of Health, Baltimore, MD 21224, USA.
Piao
Yulan
Y
Zhong
Jinhui
J
Matoba
Ryo
R
Carter
Mark G
MG
Wang
Yuxia
Y
Goldberg
Ilya
I
Ko
Minoru S H
MS
D000818:Animals; D001755:Blastocyst / Q000166:cytology* / Q000378:metabolism*; D019295:Computational Biology; D018507:Gene Expression Regulation, Developmental; D005820:Genetic Testing / Q000379:methods; D023281:Genomics / Q000379:methods / Q000706:statistics & numerical data; D017403:In Situ Hybridization / Q000295:instrumentation / Q000379:methods*; D066298:In Vitro Techniques; D051379:Mice