title: In vitro embryos production after oocytes treatment with forskolin.
abstract: The inhibition of nuclear maturation allows time for the oocyte to accumulate molecules that are important for embryonic development. Thus, the objective of this work was to evaluate the effect of blocking oocyte meiosis with the addition of forskolin, an efficient inhibitor of nuclear maturation, in in vitro maturation (IVM) medium. Forskolin was added to the IVM medium for 6 h at concentrations of 0.1 mM, 0.05 mM or 0.025 mM, then the oocytes were allowed to mature in drug-free medium for 18 h. The oocytes were assessed for the stage of nuclear maturation, the activity and distribution of mitochondria, oocyte ultrastructure, the number of viable cells and the apoptosis rate. After forskolin treatment, the oocytes were fertilized in vitro and cultured for 7 days. On day 7, the blastocyst rate, the ultrastructure, the number of intact cells and the apoptosis rate of the blastocysts were measured. No differences were observed for the stage of nuclear maturation of the oocyte, the mitochondrial activity and distribution, the blastocyst rate or total number of intact cells. However, a higher rate of apoptosis was observed in the blastocysts produced from oocytes blocked for 6 h with the higher concentration of forskolin (P < 0.05). We conclude that all the experimental groups reached the MII stage after the addition of forskolin and that the highest concentration of forskolin caused cellular degeneration without harming embryo production on the 7th day.
authors.affiliation: São Paulo State University (UNESP),School of Veterinary Medicine and Animal Science (FMVZ),Department of Animal Reproduction and Veterinary Radiology,Rubião Jr. s/n,Botucatu/SP,Brazil.
authors.forename: Daniela Martins
authors.initials: DM
authors.lastname: Paschoal
authors.affiliation: Department of Animal Reproduction and Veterinary Radiology,School of Veterinary Medicine and Animal Science (FMVZ),São Paulo State University (UNESP),Botucatu,SP,Brazil.
authors.forename: Rosiára Rosária Dias
authors.initials: RR
authors.lastname: Maziero
authors.affiliation: Department of Animal Reproduction and Veterinary Radiology,School of Veterinary Medicine and Animal Science (FMVZ),São Paulo State University (UNESP),Botucatu,SP,Brazil.
authors.forename: Mateus José
authors.initials: MJ
authors.lastname: Sudano
authors.affiliation: Department of Animal Reproduction and Veterinary Radiology,School of Veterinary Medicine and Animal Science (FMVZ),São Paulo State University (UNESP),Botucatu,SP,Brazil.
authors.forename: Midyan Daroz
authors.initials: MD
authors.lastname: Guastali
authors.affiliation: Department of Animal Reproduction and Veterinary Radiology,School of Veterinary Medicine and Animal Science (FMVZ),São Paulo State University (UNESP),Botucatu,SP,Brazil.
authors.forename: Luis Eduardo
authors.initials: LE
authors.lastname: Vergara
authors.affiliation: Department of Animal Reproduction and Veterinary Radiology,School of Veterinary Medicine and Animal Science (FMVZ),São Paulo State University (UNESP),Botucatu,SP,Brazil.
authors.forename: Letícia Ferrari
authors.initials: LF
authors.lastname: Crocomo
authors.affiliation: Department of Animal Reproduction and Veterinary Radiology,School of Veterinary Medicine and Animal Science (FMVZ),São Paulo State University (UNESP),Botucatu,SP,Brazil.
authors.forename: João Ferreira de
authors.initials: JF
authors.lastname: Lima-Neto
authors.affiliation: Department of Animal Reproduction and Veterinary Radiology,School of Veterinary Medicine and Animal Science (FMVZ),São Paulo State University (UNESP),Botucatu,SP,Brazil.
authors.forename: Luis Carlos Oña
authors.initials: LC
authors.lastname: Magalhães
authors.affiliation: Department of Animal Reproduction and Veterinary Radiology,School of Veterinary Medicine and Animal Science (FMVZ),São Paulo State University (UNESP),Botucatu,SP,Brazil.
authors.forename: Bianca Andriolo
authors.initials: BA
authors.lastname: Monteiro
authors.affiliation: Department of Animal Reproduction and Veterinary Radiology,School of Veterinary Medicine and Animal Science (FMVZ),São Paulo State University (UNESP),Botucatu,SP,Brazil.
authors.forename: Tatiana da Silva
authors.initials: Tda S
authors.lastname: Rascado
authors.affiliation: Department of Clinical Surgery and Animal Reproduction,School of Veterinary Medicine (FMVA),São Paulo State University (UNESP),Araçatuba,SP,Brazil.
authors.forename: Alício
authors.initials: A
authors.lastname: Martins
authors.affiliation: Department of Basic Science,School of Animal Science and Food Engineering (FZEA),São Paulo University (USP),Pirassununga/SP,Brazil.
authors.forename: Claudia Lima Verde
authors.initials: CL
authors.lastname: Leal
authors.affiliation: Department of Animal Reproduction and Veterinary Radiology,School of Veterinary Medicine and Animal Science (FMVZ),São Paulo State University (UNESP),Botucatu,SP,Brazil.
authors.forename: Fernanda da Cruz
authors.initials: Fda C
authors.lastname: Landim-Alvarenga
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000187:drug effects*; D002417:Cattle; D002478:Cells, Cultured; D005576:Colforsin / Q000494:pharmacology*; D047108:Embryonic Development / Q000187:drug effects; D005260:Female; D005307:Fertilization in Vitro / Q000379:methods* / Q000662:veterinary; D008297:Male; D008540:Meiosis / Q000187:drug effects; D009865:Oocytes / Q000166:cytology / Q000187:drug effects*; D013997:Time Factors; D014665:Vasodilator Agents / Q000494:pharmacology
chemical_list: D014665:Vasodilator Agents; D005576:Colforsin
keywords: Apoptosis; Forskolin; IVM; Meiosis; cAMP