title: Rapidly cleaving bovine two-cell embryos have better developmental potential and a distinctive mRNA pattern.
abstract: Mammalian embryos that rapidly reach the two-cell stage in culture have a higher probability of becoming viable blastocysts. Our goal was to separate two-cell bovine embryos based on their zygotic cleavage timing, and to assess their global mRNA levels. Following in vitro fertilization, all embryos that cleaved by 29.5 hpi (early) were cultured separately from those that divided at 46 hpi (late). The blastocyst rates were 46.1 ± 3.7% and 6.1 ± 3.4% for early- and late-cleavers, respectively (P < 0.01). Seven replicates of selected two-cell embryos were collected at each time point for microarray characterization (n = 4) and quantitative reverse-transcriptase PCR (n = 3); the rest were left in culture for blastocyst evaluation. A total of 774 and 594 probes were preferentially present in early- and late-cleaving embryos, respectively (fold change ± 1.5, P < 0.05), with important contrasts related to cell cycle, gene expression, RNA processing, and protein degradation functions. A total of 12 transcripts were assessed by quantitative PCR, of which ATM, ATR, CTNNB1, MSH6, MRE11A, PCNA, APC, CENPE, and GRB2 were in agreement with the hybridization results. Since most of these molecules are directly or indirectly associated with cell-cycle regulation, DNA damage response, and transcription control, our results strongly suggest key roles for those biological functions in mammalian preimplantation development.
authors.affiliation: Faculté des Sciences de l'Agriculture et de l'Alimentation, Département des Sciences Animales, Pavillon INAF, Centre de Recherche en Biologie de la Reproduction (CRBR), Université Laval, Québec, Canada.
authors.forename: Ernesto
authors.initials: E
authors.lastname: Orozco-Lucero
authors.forename: Isabelle
authors.initials: I
authors.lastname: Dufort
authors.forename: Claude
authors.initials: C
authors.lastname: Robert
authors.forename: Marc-André
authors.initials: MA
authors.lastname: Sirard
mesh_terms: D000818:Animals; D002417:Cattle / Q000196:embryology*; D002453:Cell Cycle / Q000235:genetics; D002970:Cleavage Stage, Ovum / Q000378:metabolism / Q000502:physiology*; D005307:Fertilization in Vitro; D046228:Microarray Analysis; D012333:RNA, Messenger / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors; D014158:Transcription, Genetic / Q000235:genetics; D059467:Transcriptome / Q000235:genetics*
chemical_list: D012333:RNA, Messenger