title: High hydrostatic pressure treatment improves the quality of in vitro-produced ovine blastocysts.
abstract: Exposure to sub-lethal hydrostatic pressure (HP) treatment is emerging as an approach to improve the general resistance of gametes and embryos to in vitro conditions. The present study was aimed to evaluate the effect of HP treatment on in vitro-produced ovine blastocysts. Experiment 1 was aimed to define optimal treatment parameters: two different HP treatments were applied to blastocysts and embryo survival was evaluated. In Experiment 2, HP parameters (40 MPa, 70 min, 38 °C) selected in Experiment 1 were used to treat blastocysts. Embryo quality was assessed and compared with untreated controls by counting total cell number, the inner cell mass (ICM) and trophectoderm (TE) cells and by evaluating nuclear picnosis. HP-treated blastocysts were processed for gene expression analysis (AQP3, ATP1A1, BAX, CDH1, HSP90β, NANOG, OCT4 and TP53) 1, 5h after the end of HP exposure. Results showed that the hatching rate of embryos treated at 40 MPa was significantly higher than that of the 60 MPa-treated group (P<0.01) and similar to untreated embryos. Blastocysts exposed at 40 MPa showed higher ICM (P<0.05) and TE (P<0.01) cell number and a lower percentage of picnotic nuclei (P<0.05) compared with the control group. Significantly lower abundance for BAX (P<0.01) and OCT4 (P<0.05) transcripts were observed in HP embryos than in the control group. In conclusion, treatment with HP improved the quality of in vitro-produced ovine blastocysts by increasing their cell number and reducing the proportion of nuclear picnosis.
authors.affiliation: Department of Veterinary Clinics and Pathology, University of Sassari, Via Vienna, 07100 Sassari, Italy. luis@uniss.it
authors.forename: Luisa
authors.initials: L
authors.lastname: Bogliolo
authors.forename: Federica
authors.initials: F
authors.lastname: Ariu
authors.forename: Giovanni
authors.initials: G
authors.lastname: Leoni
authors.forename: Stefania
authors.initials: S
authors.lastname: Uccheddu
authors.forename: Daniela
authors.initials: D
authors.lastname: Bebbere
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D001755:Blastocyst / Q000502:physiology*; D053624:Blastocyst Inner Cell Mass / Q000166:cytology; D002452:Cell Count; D002467:Cell Nucleus / Q000502:physiology; D017931:DNA Primers / Q000235:genetics; D047108:Embryonic Development / Q000502:physiology*; D005307:Fertilization in Vitro / Q000379:methods* / Q000662:veterinary*; D018507:Gene Expression Regulation, Developmental / Q000502:physiology*; D006874:Hydrostatic Pressure; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012756:Sheep / Q000196:embryology*; D016019:Survival Analysis
chemical_list: D017931:DNA Primers