title: Cloned pre-implantation mouse embryos show correct timing but altered levels of gene expression.
abstract: Mammalian embryos obtained by somatic nuclear transfer (NT) struggle to survive throughout development, encountering a number of hurdles leading to wrong functional reprogramming of the donor genome. However, despite these obstacles, some of these embryos continue their development, as if the required transcriptional functions are somehow satisfied. The amount of information gathered on the kinetics and quantitative profile of gene expression in NT pre-implantation embryos is still scarce and limited to a handful of genes described in two species, bovine and mouse. Using a single-cell sensitive semi-quantitative RT-PCR, we have compared the onset and profile of abundance of Hprt, Tsx, Bex1, Bax, Cpt2, and Oct4 genes, in in vitro fertilised and NT-derived mouse 1-cell, 2-cell, 4-cell embryos, morulae, and blastocysts. The genes analysed were activated in NT embryos at approximately the correct time compared to control embryos, indicating that the reprogramming phenomenon is developmentally regulated and that the somatic genome is quickly rearranged towards an embryonic-type of expression during the early stages of segmentation. Despite the right timing of genes onset, the high degree of variability in the number of transcripts found in NT embryos at the latest stages of pre-implantation development, suggests that genome reprogramming is incomplete and inaccurate.
authors.affiliation: Laboratorio di Biologia dello Sviluppo, Dipartimento di Biologia Animale e Centro d'Eccellenza in Biologia Applicata, Universita' degli Studi di Pavia, Pavia, Italy.
authors.forename: Vittorio
authors.initials: V
authors.lastname: Sebastiano
authors.forename: Luca
authors.initials: L
authors.lastname: Gentile
authors.forename: Silvia
authors.initials: S
authors.lastname: Garagna
authors.forename: Carlo Alberto
authors.initials: CA
authors.lastname: Redi
authors.forename: Maurizio
authors.initials: M
authors.lastname: Zuccotti
mesh_terms: D000818:Animals; D001483:Base Sequence; D001755:Blastocyst / Q000378:metabolism*; D019976:Cloning, Organism; D047108:Embryonic Development / Q000235:genetics / Q000502:physiology; D005307:Fertilization in Vitro; D018507:Gene Expression Regulation, Developmental; D051379:Mice / Q000196:embryology* / Q000235:genetics*; D008969:Molecular Sequence Data; D014158:Transcription, Genetic