Transcriptomic effects of alginate hydrogel applied to the production of bovine embryos
Giuliana de A. Ferronato,
Paola M. da S. Rosa,
Alessandra Bridi,
Angélica Camargo dos Santos,
Ricardo P. Nociti,
Marcos Roberto Chiaratti,
Felipe Perecin,
Flávio V. Meirelles,
Juliano R. Sangalli,
Juliano C. da Silveira
Affiliations
Giuliana de A. Ferronato
Faculty of Animal Science and Food Engineering, Department of Veterinary Medicine, University of São Paulo, Pirassununga, SP, Brazil
Paola M. da S. Rosa
Faculty of Animal Science and Food Engineering, Department of Veterinary Medicine, University of São Paulo, Pirassununga, SP, Brazil
Alessandra Bridi
Faculty of Animal Science and Food Engineering, Department of Veterinary Medicine, University of São Paulo, Pirassununga, SP, Brazil
Angélica Camargo dos Santos
Universidade Federal de São Carlos, Centro de Ciências Biológicas e da Saúde, Departamento de Genética e Evolução, São Carlos, SP, Brazil
Ricardo P. Nociti
Faculty of Animal Science and Food Engineering, Department of Veterinary Medicine, University of São Paulo, Pirassununga, SP, Brazil
Marcos Roberto Chiaratti
Universidade Federal de São Carlos, Centro de Ciências Biológicas e da Saúde, Departamento de Genética e Evolução, São Carlos, SP, Brazil
Felipe Perecin
Faculty of Animal Science and Food Engineering, Department of Veterinary Medicine, University of São Paulo, Pirassununga, SP, Brazil
Flávio V. Meirelles
Faculty of Animal Science and Food Engineering, Department of Veterinary Medicine, University of São Paulo, Pirassununga, SP, Brazil
Juliano R. Sangalli
Faculty of Animal Science and Food Engineering, Department of Veterinary Medicine, University of São Paulo, Pirassununga, SP, Brazil
Juliano C. da Silveira
Faculty of Animal Science and Food Engineering, Department of Veterinary Medicine, University of São Paulo, Pirassununga, SP, Brazil; Corresponding author.
In vitro-produced blastocysts are exposed to different stimuli when compared with in vivo ones. This includes the culture of in vitro embryos in a sturdy petri-dish, while in vivo embryos develop in a soft and dynamic structure. Here we hypothesized that a softer environment could differently modulate the in vitro produced embryos. To that aim, presumptive zygotes were produced by in vitro fertilization and divided into three groups: 1) Cultured in a regular Petri dish - Control (CON); 2) Cultured on top of an alginate hydrogel surface (TOP); 3) Encapsulated inside an alginate hydrogel sphere (ENC) and cultured. We observed a decrease in blastocyst rate in TOP and ENC compared with the CON. Profiling of 383 bovine miRNAs, we found 3 miRNAs involved in cell proliferation being differently modulated by the TOP and ENC groups (miR-1246; miR-1260b, and miR-541). Analyzing global levels of DNA methylation and hydroxymethylation, we observed increased levels of the two marks in the TOP group when compared with the CON and ENC systems. RNA sequencing (RNA-seq) analysis carried out using blastocysts showed alterations in several developmentally important genes among the three groups. In summary, our results indicate that in vitro embryo production was possible to achieve up to the blastocyst stage. However, with the experimental conditions used herein, the alginate hydrogels adversely affected the embryo development, which were paralleled by epigenetic and transcriptomic changes.
