04571naa a2200409 a 450000100080000000500110000800800410001902200140006002400270007410000210010124501070012226000090022950011210023852022710135965300230363065300270365365300220368065300110370265300100371365300280372365300130375170000210376470000190378570000270380470000200383170000150385170000230386670000150388970000220390470000170392670000160394370000220395970000280398170000170400970000170402677301180404310631492022-12-02       2022    bl uuuu     u00u1 u    #d  a0936-67687 a10.1111/rda.141222DOI1 aPASSOS, J. R. S.  aGlobal proteomic analysis of preimplantational ovine embryos produced in vitro.h[electronic resource]  c2022  aArticle history: Received 15 February 2022; Accepted 1 April 2022. -- Funding text - The experiments presently described were conducted at the facilities of the (Fundacion IRAUy, Montevideo, Uruguay) and at the (UBAL) of the , Uruguay. Specially, the authors thank Dr. Rosario Durán and Dr. Alejandro Leyva for kindly assisting us in the proteomic experiment. Finnacial support was provided by Fundacion IRAUy; PRONEX 02/2015 (Programa de Apoio a Núcleos de Excelência Pronex/Funcap/CNPq); the Brazilian Research Council?CNPq (grants # 313160/2017‐1 and 438773/2018‐7); Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Brazil. Instituto de Reproducción Animal Uruguay Unidad de Biotecnología en Animales de Laboratorio Institut Pasteur de Montevideo. -- Corresponding author: A. Moura, A.; Laboratório de Fisiologia e Ciências Ômicas, Departamento de Zootecnia, Universidade Federal do Ceará, Fortaleza, Brazil; email:arlindo.moura@gmail.com -- Menchaca, A.; Instituto de Reproducción Animal Uruguay, Fundación IRAUy, Montevideo, Uruguay;  mail:menchaca.alejo@gmail.com  aABSTRACT. -The present study was conducted to characterize the major proteome of preimplantation (D6) ovine embryos produced in vitro. COCs were aspirated from antral follicles (2–6 mm), matured and fertilized in vitro and cultured until day six. Proteins were ex- tracted separately from three pools of 45 embryos and separately run in SDS-PAGE. Proteins from each pool were individually subjected to in-gel digestion followed by LC-MS/MS. Three ‘raw files’ and protein  lists were produced by Pattern Lab software, but only proteins present in all three lists were used for the bioinformatics analyses. There were 2,262 proteins identified in the 6-day-old ovine  embryos, including al- bumin, zona pellucida glycoprotein 2, 3 and 4, peptidyl arginine deiminase 6, actin cytoplasmic 1, gamma-actin 1, pyruvate kinase, heat shock protein 90 and protein disulfide isomerase, among others. Major biological processes linked to the sheep embryo proteome were translation, protein transport and protein stabilization, and molecular functions, defined as ATP binding, oxygen carrier activity and oxygen bind- ing. There were 42 enriched functional clusters  according to the 2,147 genes (UniProt database). Ten selected clusters with potential association with embryo development included translation, structural constituent of ribosomes, ribosomes, nucleosomes, structural constituent of the cytoskeleton, microtubule-based process, translation initiation factor activity, regulation of translational initiation, cell body and nucleotide biosynthetic process. The most representative KEEG pathways were ribosome, oxida- tive phosphorylation, glutathione metabolism, gap junction, mineral absorption, DNA replication and cGMP-PKG signalling pathway. Analyses of functional clusters clearly showed differences associated  with the proteome of preimplantation (D6) sheep em- bryos generated after in vitro fertilization in comparison with in vivo counterparts (Sanchez et al., 2021; https://doi.org/10.1111/rda.13897), confirming that the quality of in vitro derived blastocysts are unlike those produced in vivo. The present study portrays the first comprehensive overview of the proteome of preimplantational ovine embryos grown in vitro.   © 2022 Wiley-VCH GmbH.  aEmbryo development  aIn vitro fertilization  aMass spectrometry  aOocyte  aOvine  aPLATAFORMA SALUD ANIMAL  aProteins1 aGUERREIRO, D. D.1 aOTÁVIO, K. S.1 aSANTOS-NETO, P. C. DOS1 aSOUZA-NEVES, M.1 aCUADRO, F.1 aNUÑEZ-OLIVERA, R.1 aCRISPO, M.1 aBEZERRA, M. J. B.1 aSILVA, R. F.1 aLIMA, L. F.1 aFIGUEIREDO, J. R.1 aBUSTAMANTE-FILHO, I. C.1 aMENCHACA, A.1 aMOURA, A. A.  tReproduction in Domestic Animals, 2022, Volume 57, Issue 7; pages 784-797. doi: https://doi.org/10.1111/rda.14122