title: Supplementation of IVF medium with melatonin: effect on sperm functionality and in vitro produced bovine embryos.
abstract: Gamete co-incubation generates high free radical levels surrounding growing zygotes which may impair subsequent embryo viability. Melatonin eliminates a wide variety of free radicals; hence, we tried to improve in vitro embryo production by adding melatonin to in vitro fertilisation (IVF) media in high (Exp. 1) and low concentrations (Exp. 2), and we evaluated its effect on bull sperm function during IVF co-incubation time (Exp. 3). In Experiment 1, we supplemented IVF media culture with 0.01, 0.1 and 1 mmol of melatonin, along with a no melatonin control group. In Experiment 2, melatonin levels were reduced to 10, 100 and 1000 nmol, with a no melatonin control group. In Experiment 3, spermatozoa were incubated in IVF media with melatonin (as Exp. 2) and functional parameters were analysed at 0, 4 and 18 h. In Experiment 1, only 1 mmol melatonin showed lesser blastocyst rates than control (C: 23.2 ± 6.7% versus 1 mmol: 2.0 ± 1.7%). In Experiment 2, no statistical differences were found in cleavage percentage, blastocyst percentage and total cell count for any melatonin treatment. In Experiment 3, sperm samples with 1000 nmol melatonin had a significantly higher wobbler (WOB) coefficient, a lower percentage of intact acrosomes, a lower percentage of viable spermatozoa with ROS, greater DNA fragmentation and higher DNA oxidation than controls. Total fluorescence intensity for ROS at 10 nmol melatonin was significantly greater than controls (P < 0.05). IVF media with 1 mmol melatonin is deleterious for embryo development, and in lower concentrations, it modulated sperm functionality, but had no effects on embryo production.
authors: Centro de Biotecnología de la Reproducción (BIOREN-CEBIOR), Facultad de Medicina, Universidad de La Frontera, Temuco, Chile.
authors: C
authors: C
authors: Cheuquemán
authors: Centro de Biotecnología de la Reproducción (BIOREN-CEBIOR), Facultad de Medicina, Universidad de La Frontera, Temuco, Chile.
authors: M E
authors: ME
authors: Arias
authors: Centro de Biotecnología de la Reproducción (BIOREN-CEBIOR), Facultad de Medicina, Universidad de La Frontera, Temuco, Chile.|Departamento de Ciencias Básicas, Universidad de La Frontera, Temuco, Chile.
authors: J
authors: J
authors: Risopatrón
authors: Centro de Biotecnología de la Reproducción (BIOREN-CEBIOR), Facultad de Medicina, Universidad de La Frontera, Temuco, Chile.|Departamento de Ciencias Básicas, Universidad de La Frontera, Temuco, Chile.|Departamento de Ciencias Agronómicas y Recursos Naturales, Facultad de Ciencias Agropecuarias y Forestales, Universidad de La Frontera, Temuco, Chile.
authors: R
authors: R
authors: Felmer
authors: Centro ANDROGEN, La Coruña, Spain.
authors: J
authors: J
authors: Álvarez
authors: Departamento de Medicina i Cirurgia Animals, Universitat Autònoma de Barcelona, Bellaterra, Spain.
authors: T
authors: T
authors: Mogas
authors: Centro de Biotecnología de la Reproducción (BIOREN-CEBIOR), Facultad de Medicina, Universidad de La Frontera, Temuco, Chile.|Departamento de Ciencias Preclínicas, Facultad de Medicina, Universidad de La Frontera, Temuco, Chile.
authors: R
authors: R
authors: Sánchez
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000187:drug effects; D002417:Cattle / Q000196:embryology; D002970:Cleavage Stage, Ovum / Q000187:drug effects; D003470:Culture Media; D053938:DNA Fragmentation / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D004622:Embryo, Mammalian / Q000187:drug effects*; D005307:Fertilization in Vitro / Q000379:methods / Q000662:veterinary*; D008297:Male; D008550:Melatonin / Q000627:therapeutic use*; D010084:Oxidation-Reduction / Q000187:drug effects; D017382:Reactive Oxygen Species / Q000378:metabolism; D013094:Spermatozoa / Q000187:drug effects*
chemical_list: D003470:Culture Media; D017382:Reactive Oxygen Species; D008550:Melatonin
keywords: Gamete co-incubation; in vitro fertilisation; melatonin; oxidative stress; sperm function