title: Methylation dynamics during folliculogenesis and early embryo development in sheep.
abstract: Genome-wide DNA methylation reprogramming occurs during mammalian gametogenesis and early embryogenesis. Post-fertilization demethylation of paternal and maternal genomes is considered to occur by an active and passive mechanism respectively, in most mammals but sheep; in this species no loss of methylation was observed in either pronucleus. Post-fertilization reprogramming relies on methylating and demethylating enzymes and co-factors that are stored during oocyte growth, concurrently with the re-methylation of the oocyte itself. The crucial remodelling of the oocyte epigenetic baggage often overlaps with potential interfering events such as exposure to assisted reproduction technologies or environmental changes. Here, we report a temporal analysis of methylation dynamics during folliculogenesis and early embryo development in sheep. We characterized global DNA methylation and hydroxymethylation by immunofluorescence and relatively quantified the expression of the enzymes and co-factors mainly responsible for their remodelling (DNA methyltransferases (DNMTs), ten-eleven translocation (TET) proteins and methyl-CpG-binding domain (MBD) proteins). Our results illustrate for the first time the patterns of hydroxymethylation during oocyte growth. We observed different patterns of methylation and hydroxymethylation between the two parental pronuclei, suggesting that male pronucleus undergoes active demethylation also in sheep. Finally, we describe gene-specific accumulation dynamics for methylating and demethylating enzymes during oocyte growth and observe patterns of expression associated with developmental competence in a differential model of oocyte potential. Our work contributes to the understanding of the methylation dynamics during folliculogenesis and early embryo development and improves the overall picture of early rearrangements that will originate the embryo epigenome.
authors.affiliation: Department of Veterinary MedicineUniversity of Sassari, Sassari, Italy.
authors.forename: Laura
authors.initials: L
authors.lastname: Masala
authors.affiliation: Department of Veterinary MedicineUniversity of Sassari, Sassari, Italy.
authors.forename: Giovanni Pietro
authors.initials: GP
authors.lastname: Burrai
authors.affiliation: Department of Veterinary MedicineUniversity of Sassari, Sassari, Italy.
authors.forename: Emanuela
authors.initials: E
authors.lastname: Bellu
authors.affiliation: Department of Veterinary MedicineUniversity of Sassari, Sassari, Italy.
authors.forename: Federica
authors.initials: F
authors.lastname: Ariu
authors.affiliation: Department of Veterinary MedicineUniversity of Sassari, Sassari, Italy.
authors.forename: Luisa
authors.initials: L
authors.lastname: Bogliolo
authors.affiliation: Department of Veterinary MedicineUniversity of Sassari, Sassari, Italy.
authors.forename: Sergio
authors.initials: S
authors.lastname: Ledda
authors.affiliation: Department of Veterinary MedicineUniversity of Sassari, Sassari, Italy dbebbere@uniss.it.
authors.forename: Daniela
authors.initials: D
authors.lastname: Bebbere
mesh_terms: D000818:Animals; D002467:Cell Nucleus; D019175:DNA Methylation; D004622:Embryo, Mammalian / Q000166:cytology / Q000502:physiology*; D047108:Embryonic Development / Q000235:genetics*; D005260:Female; D005306:Fertilization / Q000235:genetics*; D018507:Gene Expression Regulation, Developmental; D006080:Ovarian Follicle / Q000166:cytology / Q000502:physiology*; D012756:Sheep