Yahan Li1, Paula Tríbulo2,3, Mohammad Reza Bakhtiarizadeh4, Luiz Gustavo Siqueira5, Tieming Ji6, Rocío Melissa Rivera1, Peter James Hansen7,8. 1. Division of Animal Sciences, University of Missouri, Columbia, MO, USA. 2. Instituto de Reproduccion Animal Córdoba, X5145, Córdoba, Argentina, Spain. 3. Instituto de Ciencias Basicas Universidad Nacional de Villa Maria, X5900, Córdoba, Agentina, Spain. 4. Department of Animal and Poultry Science, College of Aburaihan, University of Tehran, Tehran, Iran. 5. Embrapa Gado de Leite, Juiz de Fora, Minas Gerais, 36038-330, Brazil. 6. Department of Statistics, University of Missouri, Columbia, MO, USA. 7. Department of Animal Sciences, University of Florida, Gainesville, FL, USA. pjhansen@ufl.edu. 8. , Gainesville, USA. pjhansen@ufl.edu.
Abstract
PURPOSE: We tested whether in vitro production (IVP) causes changes in DNA methylation in fetal liver and skeletal muscle and if exposure of cultured embryos to colony-stimulating factor 2 (CSF2) alters DNA methylation. METHODS: Female fetuses were produced by artificial insemination or transfer of an IVP embryo. Embryos were treated from days 5 to 7 after fertilization with CSF2 or vehicle. DNA methylation in fetal liver and skeletal muscle was determined by post-bisulfite adaptor tagging-based sequencing. The degree of DNA methylation for CpG sites in 50-bp windows of the promoter region 500 bp upstream of the transcriptional start site was compared between treatments. RESULTS: For liver, there were 12 genes (6% of those analyzed) in which DNA methylation was affected by treatment, with one 50-bp window per gene affected by treatment. For muscle, the degree of DNA methylation was affected by treatment for 32 windows (19% of the total windows analyzed) representing 28 distinct genes (23% of analyzed genes). For 19 of the 28 genes in muscle, the greatest deviation in DNA methylation was for the CSF2 group. CONCLUSION: Results are consistent with alterations in the methylome being one of the mechanisms by which IVP can result in altered fetal development and postnatal function in the resultant offspring. In addition, results indicate that maternally derived cell-signaling molecules can regulate the pattern of DNA methylation.
PURPOSE: We tested whether in vitro production (IVP) causes changes in DNA methylation in fetal liver and skeletal muscle and if exposure of cultured embryos to colony-stimulating factor 2 (CSF2) alters DNA methylation. METHODS: Female fetuses were produced by artificial insemination or transfer of an IVP embryo. Embryos were treated from days 5 to 7 after fertilization with CSF2 or vehicle. DNA methylation in fetal liver and skeletal muscle was determined by post-bisulfite adaptor tagging-based sequencing. The degree of DNA methylation for CpG sites in 50-bp windows of the promoter region 500 bp upstream of the transcriptional start site was compared between treatments. RESULTS: For liver, there were 12 genes (6% of those analyzed) in which DNA methylation was affected by treatment, with one 50-bp window per gene affected by treatment. For muscle, the degree of DNA methylation was affected by treatment for 32 windows (19% of the total windows analyzed) representing 28 distinct genes (23% of analyzed genes). For 19 of the 28 genes in muscle, the greatest deviation in DNA methylation was for the CSF2 group. CONCLUSION: Results are consistent with alterations in the methylome being one of the mechanisms by which IVP can result in altered fetal development and postnatal function in the resultant offspring. In addition, results indicate that maternally derived cell-signaling molecules can regulate the pattern of DNA methylation.
Entities:
Keywords:
CSF2; DNA methylation; Fetal programming; In vitro–produced embryo
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