title: Insulin influences developmental competence of bovine oocytes cultured in α-MEM plus follicle-simulating hormone.
abstract: The aim of this study was to evaluate the dose-response effect of insulin, plus follicle-simulating hormone (FSH) at a fixed concentration, in a serum-free defined culture medium (DCM) on the in vitro maturation of bovine cumulus-oocyte complexes (COCs). For oocyte nuclear maturation, the expression levels of GDF9, GLUT1, PRDX1 and HSP70.1 transcripts related to oocyte and embryo developmental competence were analysed. For in vitro maturation (IVM), cumulus-oocyte complexes from slaughterhouse ovaries were distributed into four groups based on insulin concentration added to serum-free DCM, which was composed of alpha minimum essential medium (α-MEM), as basal medium: (1) DCM control: 0 ng/ml; (2) DCM1: 1 ng/ml; (3) DCM10: 10 ng/ml; and (4) DCM100: 100 ng/ml. After IVM, the nuclear status of a sample of oocytes was analysed and the other oocytes were submitted for in vitro fertilization (IVF) and in vitro culture (IVC). Different concentrations of insulin did not affect significantly the nuclear maturation and cleavage rate (72 h post-insemination) across all groups. Blastocyst rate (192 h post-insemination) did not differ in DCM control (24.3%), DCM1 (27.0%) and DCM10 (26.3%) groups, but the DCM100 (36.1%) group showed a greater blastocyst rate (P 0.05) was observed at the different insulin concentrations. The results indicated that insulin added to DCM influenced levels of transcripts related to cellular stress (HSP70-1 and PRDX1) and oocyte competence (GDF9) in bovine oocytes and at higher concentrations enhanced blastocyst production.
authors.affiliation: Laboratory for the Study of Reproduction,Biological Institute,University of Brasília,DF,70910-900,Brazil.
authors.forename: Gustavo Bruno
authors.initials: GB
authors.lastname: Mota
authors.affiliation: Laboratory for the Study of Reproduction,Biological Institute,University of Brasília,DF,70910-900,Brazil.
authors.forename: Ingrid
authors.initials: I
authors.lastname: Oliveira e Silva
authors.affiliation: Laboratory for the Study of Reproduction,Biological Institute,University of Brasília,DF,70910-900,Brazil.
authors.forename: Danielle Kaiser
authors.initials: DK
authors.lastname: de Souza
authors.affiliation: Laboratory for the Study of Reproduction,Biological Institute,University of Brasília,DF,70910-900,Brazil.
authors.forename: Flavia
authors.initials: F
authors.lastname: Tuany
authors.affiliation: Embrapa Dairy Cattle,Juiz de Fora,MG,36038-330,Brazil.
authors.forename: Michele Munk
authors.initials: MM
authors.lastname: Pereira
authors.affiliation: Embrapa Dairy Cattle,Juiz de Fora,MG,36038-330,Brazil.
authors.forename: Luiz Sergio de Almeida
authors.initials: LS
authors.lastname: Camargo
authors.affiliation: Laboratory for the Study of Reproduction,Biological Institute,University of Brasília,DF,70910-900,Brazil.
authors.forename: Alzira Amélia Martins
authors.initials: AA
authors.lastname: Rosa e Silva
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000187:drug effects / Q000502:physiology; D002417:Cattle; D002478:Cells, Cultured; D003470:Culture Media / Q000494:pharmacology; D054885:Cumulus Cells; D004305:Dose-Response Relationship, Drug; D046149:Embryo Culture Techniques; D005260:Female; D005307:Fertilization in Vitro; D005640:Follicle Stimulating Hormone / Q000494:pharmacology*; D005786:Gene Expression Regulation / Q000187:drug effects; D051272:Glucose Transporter Type 1 / Q000235:genetics; D059471:In Vitro Oocyte Maturation Techniques / Q000379:methods; D007328:Insulin / Q000494:pharmacology*; D008297:Male; D009865:Oocytes / Q000187:drug effects* / Q000502:physiology*; D009930:Organic Chemicals / Q000494:pharmacology
chemical_list: D003470:Culture Media; D051272:Glucose Transporter Type 1; D007328:Insulin; D009930:Organic Chemicals; C420642:alpha minimal essential medium; D005640:Follicle Stimulating Hormone
keywords: Bovine; In vitro maturation; Insulin; Oocytes; Transcripts