UniGene cDNA array-based monitoring of transcriptome changes during mouse placental development
- Myriam Hemberger*,†,‡,
- James C. Cross*,
- Hans-Hilger Ropers†,
- Hans Lehrach†,
- Reinald Fundele†, and
- Heinz Himmelbauer†,‡
- *Department of Biochemistry and Molecular Biology, University of Calgary, Calgary, AB, Canada T2N 4N1; and †Max-Planck-Institut für Molekulare Genetik, D-14195 Berlin-Dahlem, Germany
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Edited by R. Michael Roberts, University of Missouri-Columbia, Columbia, MO, and approved September 14, 2001 (received for review July 27, 2001)
Abstract
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.
Footnotes
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↵ ‡ To whom reprint requests should be addressed at: Myriam Hemberger, Department of Biochemistry and Molecular Biology, University of Calgary, 3330 Hospital Drive Northwest, Calgary, AB, Canada T2N 4N1. E-mail: mhemberg{at}ucalgary.ca; or Heinz Himmelbauer, Max-Planck-Institut für Molekulare Genetik, Ihnestrasse 73, D-14195 Berlin, Germany. E-mail: himmelbauer{at}molgen.mpg.de.
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This paper was submitted directly (Track II) to the PNAS office.
- Abbreviation:
- EST,
- expressed sequence tag
- Copyright © 2001, The National Academy of Sciences
