cDNA microarray analysis of bovine embryo gene expression profiles during the pre-implantation period.
AuthorUshizawa, K; Herath, CB; Kaneyama, K; Shiojima, S; Hirasawa, A; Takahashi, T; Imai, K; Ochiai, K; Tokunaga, T; Tsunoda, Y; ...
Source TitleReproductive Biology and Endocrinology
PublisherSpringer Science and Business Media LLC
University of Melbourne Author/sHerath, Chandana
AffiliationMedicine (Austin & Northern Health)
Document TypeJournal Article
CitationsUshizawa, K., Herath, C. B., Kaneyama, K., Shiojima, S., Hirasawa, A., Takahashi, T., Imai, K., Ochiai, K., Tokunaga, T., Tsunoda, Y., Tsujimoto, G. & Hashizume, K. (2004). cDNA microarray analysis of bovine embryo gene expression profiles during the pre-implantation period.. Reprod Biol Endocrinol, 2 (1), pp.77-. https://doi.org/10.1186/1477-7827-2-77.
Access StatusOpen Access
Open Access at PMChttp://www.ncbi.nlm.nih.gov/pmc/articles/PMC535809
BACKGROUND: After fertilization, embryo development involves differentiation, as well as development of the fetal body and extra-embryonic tissues until the moment of implantation. During this period various cellular and molecular changes take place with a genetic origin, e.g. the elongation of embryonic tissues, cell-cell contact between the mother and the embryo and placentation. To identify genetic profiles and search for new candidate molecules involved during this period, embryonic gene expression was analyzed with a custom designed utero-placental complementary DNA (cDNA) microarray. METHODS: Bovine embryos on days 7, 14 and 21, extra-embryonic membranes on day 28 and fetuses on days 28 were collected to represent early embryo, elongating embryo, pre-implantation embryo, post-implantation extra-embryonic membrane and fetus, respectively. Gene expression at these different time points was analyzed using our cDNA microarray. Two clustering algorithms such as k-means and hierarchical clustering methods identified the expression patterns of differentially expressed genes across pre-implantation period. Novel candidate genes were confirmed by real-time RT-PCR. RESULTS: In total, 1,773 individual genes were analyzed by complete k-means clustering. Comparison of day 7 and day 14 revealed most genes increased during this period, and a small number of genes exhibiting altered expression decreased as gestation progressed. Clustering analysis demonstrated that trophoblast-cell-specific molecules such as placental lactogens (PLs), prolactin-related proteins (PRPs), interferon-tau, and adhesion molecules apparently all play pivotal roles in the preparation needed for implantation, since their expression was remarkably enhanced during the pre-implantation period. The hierarchical clustering analysis and RT-PCR data revealed new functional roles for certain known genes (dickkopf-1, NPM, etc) as well as novel candidate genes (AW464053, AW465434, AW462349, AW485575) related to already established trophoblast-specific genes such as PLs and PRPs. CONCLUSIONS: A large number of genes in extra-embryonic membrane increased up to implantation and these profiles provide information fundamental to an understanding of extra-embryonic membrane differentiation and development. Genes in significant expression suggest novel molecules in trophoblast differentiation.
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