UniGene cDNA array-based monitoring of transcriptome changes during mouse placental development.
The placenta is a highly specialized organ essential for embryonic growth and development. Here, we have applied cDNA subtraction between extraembryonic tissues of early- (day 7.5 of gestation) and late-stage embryos (day 17.5) to generate stage-specific cDNA pools that were used for screening of high-density mouse UniGene cDNA arrays containing 25,000 clones. A total of 638 clones were identified, 488 with the e7.5-specific probe and 150 with the e17.5-specific probe. Importantly, 363/638 (56.9%) of the hybridizing clones were not known to be expressed during placental development before. Differential regulation was confirmed by Northern blot and in situ hybridization for a total of 44/44 of positive clones. Thus, this combination of cDNA subtraction and array hybridization was highly successful for identification of genes expressed and regulated during placental development. These included growth factors and receptors, components of the transcriptional and translational machinery, cell cycle regulators, molecular chaperones, and cytoskeletal elements. The extensive in situ hybridization analysis revealed extraembryonic structures with a high density of differentially expressed genes, most strikingly the ectoplacental cone and the spongiotrophoblast. This large-scale identification of genes regulated during placentogenesis is extremely useful to further elucidate the molecular basis of extraembryonic development.
Hemberger
M
M
Department of Biochemistry and Molecular Biology, University of Calgary, Calgary, AB, Canada T2N 4N1. mhemberg@ucalgary.ca
Cross
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Ropers
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Lehrach
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Fundele
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Himmelbauer
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D000818:Animals; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics*; D005786:Gene Expression Regulation; D017403:In Situ Hybridization; D051379:Mice; D008815:Mice, Inbred Strains; D020411:Oligonucleotide Array Sequence Analysis; D010920:Placenta / Q000378:metabolism; D010929:Placentation; D014158:Transcription, Genetic
D018076:DNA, Complementary