title: In vivo-derived horse blastocysts show transcriptional upregulation of developmentally important genes compared with in vitro-produced horse blastocysts.
abstract: In vitro-produced (IVP) equine blastocysts can give rise to successful pregnancies, but their morphology and developmental rate differ from those of in vivo-derived equine blastocysts. The aim of the present study was to evaluate this difference at the genetic level. Suppression subtractive hybridisation (SSH) was used to construct a cDNA library enriched for transcripts preferentially expressed in in vivo-derived equine blastocysts compared with IVP blastocysts. Of the 62 different genes identified in this way, six genes involved in embryonic development (BEX2, FABP3, HSP90AA1, MOBKL3, MCM7 and ODC) were selected to confirm this differential expression by reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR). Using RT-qPCR, five genes were confirmed to be significantly upregulated in in vivo-derived blastocysts (i.e. FABP3, HSP90AA1 (both P<0.05), ODC, MOBKL3 and BEX2 (P<0.005 for all three)), confirming the results of the SSH. There was no significant difference in MCM7 expression between IVP and in vivo-derived blastocysts. In conclusion, five genes that are transcriptionally upregulated in in vivo-derived equine blastocysts compared with IVP blastocysts have been identified. Because of their possible importance in embryonic development, the expression of these genes can be used as a marker to evaluate in vitro embryo production systems in the horse.
authors.affiliation: Department of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, B-9820 Merelbeke, Belgium. katrien.smits@ugent.be
authors.forename: Katrien
authors.initials: K
authors.lastname: Smits
authors.forename: Karen
authors.initials: K
authors.lastname: Goossens
authors.forename: Ann
authors.initials: A
authors.lastname: Van Soom
authors.forename: Jan
authors.initials: J
authors.lastname: Govaere
authors.forename: Maarten
authors.initials: M
authors.lastname: Hoogewijs
authors.forename: Luc J
authors.initials: LJ
authors.lastname: Peelman
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics; D000818:Animals; D001755:Blastocyst / Q000378:metabolism*; D050556:Fatty Acid-Binding Proteins / Q000235:genetics; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary*; D018507:Gene Expression Regulation, Developmental; D018841:HSP90 Heat-Shock Proteins / Q000235:genetics; D006736:Horses / Q000196:embryology*; D009419:Nerve Tissue Proteins / Q000235:genetics; D009693:Nucleic Acid Hybridization; D009955:Ornithine Decarboxylase / Q000235:genetics; D011247:Pregnancy; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015854:Up-Regulation
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D050556:Fatty Acid-Binding Proteins; D018841:HSP90 Heat-Shock Proteins; D009419:Nerve Tissue Proteins; D009955:Ornithine Decarboxylase