title: Proteome analysis of early lineage specification in bovine embryos.
abstract: During mammalian embryo development, the zygote undergoes embryonic cleavage in the oviduct and reaches the uterus at the morula stage, when compaction and early lineage specification take place. To increase knowledge about the associated changes of the embryonic protein repertoire, we performed a comprehensive proteomic analysis of in vitro produced bovine morulae and blastocysts (six biological replicates), using an iTRAQ-based approach. A total of 560 proteins were identified of which 502 were quantified. The abundance of 140 proteins was significantly different between morulae and blastocysts, among them nucleophosmin (NPM1), eukaryotic translation initiation factor 5A-1 (EIF5A), receptor of activated protein kinase C 1 (GNB2L1/RACK1), and annexin A6 (ANXA6) with increased, and glutathione S-transferase mu 3 (GSTM3), peroxiredoxin 2 (PRDX2), and aldo-keto reductase family 1 member B1 (AKR1B1) with decreased abundance in blastocysts. Seventy-three percent of abundance altered proteins increased, reflecting an increase of translation activity in this period. This is further supported by an increase in the abundance of proteins involved in the translation machinery and the synthesis of ATP. Additionally, a complementary 2D saturation DIGE analysis led to the detection of protein isoforms, e.g. of GSTM3 and PRDX2, relevant for this period of mammalian development, and exemplarily verified the results of the iTRAQ approach. In summary, our systematic differential proteome analysis of bovine morulae and blastocysts revealed new molecular correlates of early lineage specification and differentiation events during bovine embryogenesis.
authors.affiliation: Laboratory for Functional Genome Analysis (LAFUGA), Gene Center, Ludwig-Maximilians-Universität München, Munich, Germany.
authors.forename: Myriam
authors.initials: M
authors.lastname: Demant
authors.forename: Daniela R
authors.initials: DR
authors.lastname: Deutsch
authors.forename: Thomas
authors.initials: T
authors.lastname: Fröhlich
authors.forename: Eckhard
authors.initials: E
authors.lastname: Wolf
authors.forename: Georg J
authors.initials: GJ
authors.lastname: Arnold
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000737:chemistry / Q000378:metabolism*; D002417:Cattle; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D007553:Isotope Labeling; D009028:Morula / Q000737:chemistry / Q000378:metabolism*; D060066:Protein Interaction Maps / Q000502:physiology*; D011506:Proteins / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D020543:Proteome / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D040901:Proteomics; D053719:Tandem Mass Spectrometry
chemical_list: D011506:Proteins; D020543:Proteome
keywords: Animal proteomics; Blastocyst; Embryo; Pluripotency; Saturation DIGE; iTRAQ