title: Effects of high hydrostatic pressure on genomic expression profiling of porcine parthenogenetic activated and cloned embryos.
abstract: Handmade cloning (HMC) has been used to generate transgenic pigs for biomedical research. Recently, we found that parthenogenetic activation (PA) of porcine oocytes and improved HMC efficiency could be achieved by treatment with sublethal high hydrostatic pressure (HHP). However, the molecular mechanism underlying the effects of HHP treatment on embryonic development is poorly understood and so was investigated in the present study. Thus, in the present study, we undertook genome-wide gene expression analysis in HHP-treated and untreated oocytes, as well as in 4-cell and blastocyst stage embryos derived by PA or HMC. Hierarchical clustering depicted stage-specific genomic expression profiling. At the 4-cell and blastocyst stages, 103 and 163 transcripts were differentially expressed between the HMC and PA embryos, respectively (P<0.05). These transcripts are predominantly involved in regulating cellular differentiation, gene expression and cell-to-cell signalling. We found that 44 transcripts were altered by HHP treatment, with most exhibiting lower expression in HHP-treated oocytes. Genes involved in embryonic development were prominent among the transcripts affected by HHP. Two of these genes (INHBB and ME3) were further validated by quantitative reverse transcription-polymerase chain reaction. We also observed that HHP treatment activated expression of the imprinting gene DLX5 in 4-cell PA embryos. In conclusion, our genomic expression profiling data suggest that HHP alters the RNA constitution in porcine oocytes and affects the expression of imprinting genes during embryonic development.
authors.affiliation: Department of Biomedicine, Aarhus University, Wilhelm Meyer Alle 4, 8000, Aarhus C, Denmark.
authors.forename: Lin
authors.initials: L
authors.lastname: Lin
authors.affiliation: Department of Biomedicine, Aarhus University, Wilhelm Meyer Alle 4, 8000, Aarhus C, Denmark.
authors.forename: Yonglun
authors.initials: Y
authors.lastname: Luo
authors.affiliation: Department of Animal Science, Aarhus University, Blichers Alle 20, 8830, Tjele, Denmark.
authors.forename: Peter
authors.initials: P
authors.lastname: Sørensen
authors.affiliation: Department of Biomedicine, Aarhus University, Wilhelm Meyer Alle 4, 8000, Aarhus C, Denmark.
authors.forename: Helle
authors.initials: H
authors.lastname: Prætorius
authors.affiliation: BGI/HuaDa, Beishan Road 10, 518000, Shenzhen, China.
authors.forename: Gabor
authors.initials: G
authors.lastname: Vajta
authors.affiliation: Department of Animal Science, Aarhus University, Blichers Alle 20, 8830, Tjele, Denmark.
authors.forename: Henrik
authors.initials: H
authors.lastname: Callesen
authors.affiliation: Department of Animal Breeding and Genetics, Szent István University, István u. 2, Budapest, 1078, Hungary.
authors.forename: Csaba
authors.initials: C
authors.lastname: Pribenszky
authors.affiliation: Department of Biomedicine, Aarhus University, Wilhelm Meyer Alle 4, 8000, Aarhus C, Denmark.
authors.forename: Lars
authors.initials: L
authors.lastname: Bolund
authors.affiliation: Department of Molecular Biology and Genetics, Center for Quantitative Genetics and Genomics, Aarhus University, Blichers Alle 20, 8830, Tjele, Denmark.
authors.forename: Torsten Nygård
authors.initials: TN
authors.lastname: Kristensen
mesh_terms: D000367:Age Factors; D000818:Animals; D019976:Cloning, Organism / Q000379:methods / Q000662:veterinary*; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental / Q000235:genetics / Q000502:physiology*; D018392:Genomic Imprinting; D006874:Hydrostatic Pressure; D046228:Microarray Analysis; D010312:Parthenogenesis / Q000235:genetics / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013552:Swine / Q000196:embryology*