title: Transcriptome Analysis of Pig In Vivo, In Vitro-Fertilized, and Nuclear Transfer Blastocyst-Stage Embryos Treated with Histone Deacetylase Inhibitors Postfusion and Activation Reveals Changes in the Lysosomal Pathway.
abstract: Genetically modified pigs are commonly created via somatic cell nuclear transfer (SCNT). Treatment of reconstructed embryos with histone deacetylase inhibitors (HDACi) immediately after activation improves cloning efficiency. The objective of this experiment was to evaluate the transcriptome of SCNT embryos treated with suberoylanilide hydroxamic acid (SAHA), 4-iodo-SAHA (ISAHA), or Scriptaid as compared to untreated SCNT, in vitro-fertilized (IVF), and in vivo (IVV) blastocyst-stage embryos. SAHA (10 μM) had the highest level of blastocyst development at 43.9%, and all treatments except 10 μM ISAHA had the same percentage of blastocyst development as Scriptaid (p<0.05). Two treatments, 1.0 μM ISAHA and 1.0 μM SAHA, had higher mean cell number than No HDACi treatment (p<0.021). Embryo transfers performed with 10 μM SAHA- and 1 μM ISAHA-treated embryos resulted in the birth of healthy piglets. GenBank accession numbers from up- and downregulated transcripts were loaded into the Database for Annotation, Visualization and Integrated Discovery to identify enriched biological themes. HDACi treatment yielded the highest enrichment for transcripts within the Kyoto Encyclopedia of Genes and Genomes (KEGG) Pathway, lysosome. The mean intensity of LysoTracker was lower in IVV embryos compared to IVF and SCNT embryos (p<0.0001). SAHA and ISAHA can successfully be used to create healthy piglets from SCNT.
authors.affiliation: 1 Division of Animal Sciences, National Swine Research and Resource Center, University of Missouri , Columbia, MO, 65211.
authors.forename: Kristin M
authors.initials: KM
authors.lastname: Whitworth
authors.affiliation: 1 Division of Animal Sciences, National Swine Research and Resource Center, University of Missouri , Columbia, MO, 65211.
authors.forename: Jiude
authors.initials: J
authors.lastname: Mao
authors.affiliation: 2 Department of Animal and Poultry Science, Virginia Tech , Blacksburg, VA, 24061.
authors.forename: Kiho
authors.initials: K
authors.lastname: Lee
authors.affiliation: 3 Informatics Research Core Facility, University of Missouri , Columbia, MO, 65211.
authors.forename: William G
authors.initials: WG
authors.lastname: Spollen
authors.affiliation: 1 Division of Animal Sciences, National Swine Research and Resource Center, University of Missouri , Columbia, MO, 65211.
authors.forename: Melissa S
authors.initials: MS
authors.lastname: Samuel
authors.affiliation: 1 Division of Animal Sciences, National Swine Research and Resource Center, University of Missouri , Columbia, MO, 65211.
authors.forename: Eric M
authors.initials: EM
authors.lastname: Walters
authors.affiliation: 1 Division of Animal Sciences, National Swine Research and Resource Center, University of Missouri , Columbia, MO, 65211.
authors.forename: Lee D
authors.initials: LD
authors.lastname: Spate
authors.affiliation: 1 Division of Animal Sciences, National Swine Research and Resource Center, University of Missouri , Columbia, MO, 65211.
authors.forename: Randall S
authors.initials: RS
authors.lastname: Prather
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000187:drug effects* / Q000378:metabolism; D005260:Female; D005307:Fertilization in Vitro; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology*; D006877:Hydroxamic Acids / Q000032:analysis / Q000494:pharmacology; D006898:Hydroxylamines / Q000494:pharmacology; D008247:Lysosomes / Q000187:drug effects / Q000201:enzymology; D053652:Nuclear Transfer Techniques; D011804:Quinolines / Q000494:pharmacology; D034421:Sus scrofa / Q000196:embryology*; D059467:Transcriptome; D000077337:Vorinostat
chemical_list: D056572:Histone Deacetylase Inhibitors; D006877:Hydroxamic Acids; D006898:Hydroxylamines; D011804:Quinolines; C410733:scriptaid; D000077337:Vorinostat