title: Global proteomic characterization of uterine histotroph recovered from beef heifers yielding good quality and degenerate day 7 embryos.
abstract: The objective was to analyze the proteomic composition of uterine flushes collected from beef heifers on day 7 after insemination. Estrus was synchronized in crossbred beef heifers by using a protocol with a controlled intravaginal drug releasing device. Heifers detected in standing estrus (within 24-48 h after removal of controlled intravaginal drug releasing device) were inseminated (estrus = day 0) with frozen-thawed semen from a single ejaculate of a bull with proven fertility. Heifers from which an embryo was recovered (after slaughter on day 7) were classified as either having a viable embryo (morula/blastocyst stage) or a degenerate embryo (arrested at the 2- to 16-cell stage). The overall recovery rate (viable and degenerate combined) was 64%. Global liquid chromatography coupled to tandem mass spectrometry proteomic analysis of the histotroph collected identified 40 high-confidence proteins present on day 7; 26 proteins in the viable group, 10 in the degenerate group, and 4 shared between both groups. Five proteins (platelet-activating factor acetylhydrolase IB subunit γ [PAFAH1B3], tubulin α-1D chain, tubulin β-4A chain, cytochrome C, and dihydropyrimidinase-related protein-2) were unique or more abundant in the histotroph collected from animals with a viable embryo, and 1 protein (S100-A4) was more abundant in the histotroph collected from animals with a degenerate embryo. Of interest, PAFAH1B3, detected only in histotroph from the group yielding viable embryos, belongs to the group of platelet-activating factors that are known to be important for the development of the pre-implantation embryo in other species. To our knowledge this is the first report of PAFAH1B3 in relation to bovine early embryonic development.
authors.affiliation: School of Veterinary Medicine, University College Dublin, Belfield, Dublin 4, Ireland. Electronic address: marijke.beltman@ucd.ie.
authors.forename: M E
authors.initials: ME
authors.lastname: Beltman
authors.affiliation: School of Veterinary Medicine, University College Dublin, Belfield, Dublin 4, Ireland; Teagasc, Animal and Bioscience Research Centre, Animal & Grassland Research and Innovation Centre, Athenry, County Galway, Ireland.
authors.forename: M P
authors.initials: MP
authors.lastname: Mullen
authors.affiliation: Conway Institute, University College Dublin, Belfield, Dublin 4, Ireland.
authors.forename: G
authors.initials: G
authors.lastname: Elia
authors.affiliation: Conway Institute, University College Dublin, Belfield, Dublin 4, Ireland; National Institute for Bioprocessing Research & Training, Blackrock, County Dublin, Ireland.
authors.forename: M
authors.initials: M
authors.lastname: Hilliard
authors.affiliation: Teagasc, Animal and Bioscience Research Centre, Animal & Grassland Research and Innovation Centre, Athenry, County Galway, Ireland.
authors.forename: M G
authors.initials: MG
authors.lastname: Diskin
authors.affiliation: School of Agriculture and Food Science, University College Dublin, Belfield, Dublin 4, Ireland.
authors.forename: A C
authors.initials: AC
authors.lastname: Evans
authors.affiliation: School of Veterinary Medicine, University College Dublin, Belfield, Dublin 4, Ireland; Teagasc, Animal and Bioscience Research Centre, Animal & Grassland Research and Innovation Centre, Athenry, County Galway, Ireland; Conway Institute, University College Dublin, Belfield, Dublin 4, Ireland.
authors.forename: M A
authors.initials: MA
authors.lastname: Crowe
mesh_terms: D000818:Animals; D002417:Cattle / Q000378:metabolism*; D002853:Chromatography, Liquid / Q000662:veterinary; D047108:Embryonic Development / Q000502:physiology*; D005260:Female; D063990:Gene Ontology; D011247:Pregnancy; D011374:Progesterone / Q000097:blood; D040901:Proteomics / Q000379:methods; D021241:Spectrometry, Mass, Electrospray Ionization / Q000662:veterinary; D014599:Uterus / Q000378:metabolism*
chemical_list: D011374:Progesterone
keywords: Embryo development; Histotroph; Proteomics