title: Effects of FGF10 on bovine oocyte meiosis progression, apoptosis, embryo development and relative abundance of developmentally important genes in vitro.
abstract: Fibroblast growth factor (FGF10) acts at the cumulus oocyte complex, increasing the expression of cumulus cell expansion-related genes and oocyte competency genes. We tested the hypothesis that addition of FGF10 to the maturation medium improves oocyte maturation, decreases the percentage of apoptotic oocytes and increases development to the blastocyst stage while increasing the relative abundance of developmentally important genes (COX2, CDX2 and PLAC8). In all experiments, oocytes were matured for 22 h in TCM-199 supplemented with 0, 2.5, 10 or 50 ng/ml FGF10. In Experiment 1, after maturation, oocytes were stained with Hoechst to evaluate meiosis progression (metaphase I, intermediary phases and extrusion of the first polar body) and submitted to the TUNEL assay to evaluate apoptosis. In Experiment 2, oocytes were fertilized and cultured to the blastocyst stage. Blastocysts were frozen for analysis of COX2, CDX2 and PLAC8 relative abundance. In Experiment 1, 2.5 ng/ml FGF10 increased (p < 0.05) the percentage of oocytes with extrusion of the first polar body (35%) compared to 0, 10 and 50 ng/ml FGF10 (21, 14 and 12%, respectively) and FGF10 decreased the percentage of oocytes that were TUNEL positive in all doses studied. In Experiment 2, there was no difference in the percentage of oocytes becoming blastocysts between treatments and control. Real-time RT-PCR showed a tendency of 50 ng/ml FGF10 to increase the relative abundance of COX2 and PLAC8 and of 10 ng/ml FGF10 to increase CDX2. In conclusion, the addition of FGF10 to the oocyte maturation medium improves oocyte maturation in vitro, decreases the percentage of apoptotic oocytes and tends to increase the relative abundance of developmentally important genes.
authors.affiliation: Department of Pharmacology, Institute of Biosciences, São Paulo State University (UNESP), Botucatu, SP, Brazil.
authors.forename: R F
authors.initials: RF
authors.lastname: Pomini Pinto
authors.forename: P K
authors.initials: PK
authors.lastname: Fontes
authors.forename: B
authors.initials: B
authors.lastname: Loureiro
authors.forename: A C
authors.initials: AC
authors.lastname: Sousa Castilho
authors.forename: J
authors.initials: J
authors.lastname: Sousa Ticianelli
authors.forename: E
authors.initials: E
authors.lastname: Montanari Razza
authors.forename: R A
authors.initials: RA
authors.lastname: Satrapa
authors.forename: J
authors.initials: J
authors.lastname: Buratini
authors.forename: C
authors.initials: C
authors.lastname: Moraes Barros
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D001755:Blastocyst / Q000737:chemistry / Q000502:physiology; D000071616:CDX2 Transcription Factor; D002417:Cattle; D003470:Culture Media; D054885:Cumulus Cells / Q000502:physiology; D051546:Cyclooxygenase 2 / Q000235:genetics; D046149:Embryo Culture Techniques / Q000662:veterinary; D047108:Embryonic Development / Q000187:drug effects*; D005260:Female; D005307:Fertilization in Vitro / Q000187:drug effects; D051526:Fibroblast Growth Factor 10 / Q000008:administration & dosage / Q000494:pharmacology*; D050437:Genes, Developmental; D018398:Homeodomain Proteins / Q000235:genetics; D020287:In Situ Nick-End Labeling; D059471:In Vitro Oocyte Maturation Techniques; D008540:Meiosis / Q000187:drug effects*; D009865:Oocytes / Q000737:chemistry / Q000166:cytology*; D011257:Pregnancy Proteins / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D015534:Trans-Activators / Q000235:genetics
chemical_list: D000071616:CDX2 Transcription Factor; D003470:Culture Media; D051526:Fibroblast Growth Factor 10; D018398:Homeodomain Proteins; D011257:Pregnancy Proteins; D012333:RNA, Messenger; D015534:Trans-Activators; D051546:Cyclooxygenase 2