title: Regulation of blastocyst stage gene expression and outgrowth interferon tau activity of somatic cell clone aggregates.
abstract: The inefficiency of mammalian somatic cell cloning is associated with abnormal gene expression presumably caused by errors in reprogramming of the transplanted genome. In the mouse, aggregation of four-cell stage clones leads to an improvement of both gene expression and development. To determine whether clone-clone aggregation at postgenomic activation stages influences gene expression in bovine clones, we profiled, in single and aggregated embryos at the blastocyst stage, expression of developmentally relevant genes namely Oct4, Dnmt1, Dnmt3, Glut1, Glut3, and a housekeeping gene, Poly(A) polymerase (PolyA) by real-time RT-PCR. Compared to embryos generated by in vitro fertilization (IVF), individual clones more frequently exhibited transcript levels that were more than twofold higher or lower than the average value of IVF embryos. This was observed less often in clone aggregates for Oct4, Dnmt1, Dnmt3, and PolyA, but not for Glut1 and Glut3. The analysis of interferon tau bioactivity as a marker of trophectoderm function in blastocyst outgrowths showed that both single clones and clone aggregates have less extraembryonic potential in vitro compared to IVF embryos, with no apparent consequence of aggregation. These findings indicate that aggregation of bovine clones with each other at later cleavage stages can change gene expression patterns at preimplantation stages, but does not rescue trophectoderm function in vitro.
authors.affiliation: Center for Animal Transgenesis and Germ Cell Research, University of Pennsylvania, Kennett Square, Pennsylvania 19348, USA.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Kurosaka
authors.forename: Sigrid
authors.initials: S
authors.lastname: Eckardt
authors.forename: Alan D
authors.initials: AD
authors.lastname: Ealy
authors.forename: K John
authors.initials: KJ
authors.lastname: McLaughlin
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000166:cytology*; D002417:Cattle; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D019976:Cloning, Organism / Q000379:methods*; D017931:DNA Primers / Q000737:chemistry; D004475:Ectoderm / Q000378:metabolism; D010064:Embryo Implantation; D005307:Fertilization in Vitro; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D009865:Oocytes / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D016875:tau Proteins / Q000378:metabolism*
chemical_list: D017931:DNA Primers; D012333:RNA, Messenger; D016875:tau Proteins