title: Supplementation of bovine embryo culture medium with L-arginine improves embryo quality via nitric oxide production.
abstract: Nitric oxide (NO) is a cell-signaling molecule that regulates a variety of molecular pathways. We investigated the role of NO during preimplantation embryonic development by blocking its production with an inhibitor or supplementing in vitro bovine embryo cultures with its natural precursor, L-arginine, over different periods. Endpoints evaluated included blastocyst rates, development kinetics, and embryo quality. Supplementation with the NO synthase inhibitor N-Nitro-L-arginine-methyl ester (L-NAME) from Days 1 to 8 of culture decreased blastocyst (P < 0.05) and hatching (P < 0.05) rates. When added from Days 1 to 8, 50 mM L-arginine decreased blastocyst rates (P < 0.001); in contrast, when added from Days 5 to 8, 1 mM L-arginine improved embryo hatching rates (P < 0.05) and quality (P < 0.05) as well as increased POU5F1 gene expression (P < 0.05) as compared to the untreated control. Moreover, NO levels in the medium during this culture period positively correlated with the increased embryo hatching rates and quality (P < 0.05). These data suggest exerts its positive effects during the transition from morula to blastocyst stage, and that supplementing the embryo culture medium with L-arginine favors preimplantation development of bovine embryos.
authors.affiliation: Federal Rural University of Amazon, Capitão-Poço, Pará, Brazil.
authors.forename: Priscila Di Paula Bessa
authors.initials: PD
authors.lastname: Santana
authors.forename: Thiago Velasco Guimarães
authors.initials: TV
authors.lastname: Silva
authors.forename: Nathália Nogueira
authors.initials: NN
authors.lastname: da Costa
authors.forename: Bruno Barauna
authors.initials: BB
authors.lastname: da Silva
authors.forename: Timothy Frederick
authors.initials: TF
authors.lastname: Carter
authors.forename: Marcela da Silva
authors.initials: Mda S
authors.lastname: Cordeiro
authors.forename: Bruno José Martins
authors.initials: BJ
authors.lastname: da Silva
authors.forename: Simone do Socorro Damasceno
authors.initials: Sdo S
authors.lastname: Santos
authors.forename: Anderson Manoel
authors.initials: AM
authors.lastname: Herculano
authors.forename: Paulo Roberto
authors.initials: PR
authors.lastname: Adona
authors.forename: Otávio Mitio
authors.initials: OM
authors.lastname: Ohashi
authors.forename: Moysés dos Santos
authors.initials: Mdos S
authors.lastname: Miranda
mesh_terms: D000818:Animals; D001120:Arginine / Q000494:pharmacology*; D001755:Blastocyst / Q000166:cytology / Q000378:metabolism*; D002417:Cattle; D003470:Culture Media / Q000737:chemistry / Q000494:pharmacology; D046149:Embryo Culture Techniques; D047108:Embryonic Development / Q000187:drug effects*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D019331:NG-Nitroarginine Methyl Ester / Q000494:pharmacology; D009569:Nitric Oxide / Q000378:metabolism*
chemical_list: D003470:Culture Media; D004791:Enzyme Inhibitors; D009569:Nitric Oxide; D001120:Arginine; D019331:NG-Nitroarginine Methyl Ester