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Consequences of Nitric Oxide Synthase Inhibition During Bovine Oocyte Maturation on Meiosis and Embryo Development
Authors:KRL Schwarz  PRL Pires  THC de Bem  PR Adona   CLV Leal
Affiliation:Departamento de Ciências Básicas, Faculdade de Zootecnia e Engenharia de Alimentos, Universidade de São Paulo, Pirassununga-SP, Brazil
Abstract:The importance of nitric oxide synthase (NOS) in bovine oocyte maturation was investigated. Oocytes were in vitro matured with the NOS inhibitor Nw- l -nitro-arginine methyl-ester (10−7, 10−5 and 10−3  m l -NAME) and metaphase II (MII) rates and embryo development and quality were assessed. The effect of l -NAME (10−7  m ) during pre-maturation and/or maturation on embryo development and quality was also assessed. l -NAME decreased MII rates (78–82%, p < 0.05) when compared with controls without l -NAME (96%). Cleavage (77–88%, p > 0.05), Day 7 blastocyst rates (34–42%, p > 0.05) and total cell numbers in blastocysts were similar for all groups (146–171 cells, p > 0.05). Day 8 blastocyst TUNEL positive cells (3–4 cells) increased with l -NAME treatment (p < 0.05). For oocytes cultured with l -NAME during pre-maturation and/or maturation, Day 8 blastocyst development (26–34%) and Day 9 hatching rates (15–22%) were similar (p > 0.05) to controls pre-matured and matured without NOS inhibition (33 and 18%, respectively), while total cell numbers (Day 9 hatched blastocysts) increased (264–324 cells, p < 0.05) when compared with the controls (191 cells). TUNEL positive cells increased when NOS was inhibited only during the maturation period (8 cells, p < 0.05) when compared with the other groups (3–4 cells). NO may be involved in meiosis progression to MII and its deficiency during maturation increases apoptosis in embryos produced in vitro . Nitric oxide synthase inhibition during pre-maturation and/or maturation affects embryo quality.
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