Banner
Current IssueEditorial BoardOnline First ArticlesArchive
Current IssueEditorial BoardOnline First ArticlesArchive

volume 48 issue 3 (june 2014) : 221-226,   Doi: 10.5958/j.0976-0555.48.3.047

THE LEVELS OF DNA METHYLATION OF SHEEP CLONED EMBRYOS IN DIFFERENT DEVELOPMENT STAGES

L
L.B. Ma*
X
X.Y. He
1School of Mathematics, Physics and Biological Engineering, Inner Mongolia University of Science & Technology, Baotou, Inner Mongolia, China
Cite article:- Ma* L.B., He X.Y. (2025). THE LEVELS OF DNA METHYLATION OF SHEEP CLONED EMBRYOS IN DIFFERENT DEVELOPMENT STAGES. Indian Journal of Animal Research. 48(3): 221-226. doi: 10.5958/j.0976-0555.48.3.047.

ABSTRACT

DNA methylation is a kind of epigenetic modification. During the early development of cloned embryos, parental genomic DNA undergoes active- and passive demethylation as well as low level of DNA methylation in cloned embryos at morula stage. In this study, sheep skin fibroblasts were used as nuclear donors, and sheep oocytes were used as nuclear recipient, sheep cloned embryos were constructed by the technology of somatic cell nuclear transfer, the development ability of cloned embryos was evaluated. Moreover, the dynamic change in DNA methylation level was also examined by the technology of methyl sensitive amplified polymorphism. The results showed that sheep cloned embryo had the ability to develop to blastocyst stage, and the level of DNA methylation decreased from nonactivated 1-cell stage to 4-cell stage, and then increased onward to blastocyst stage; while the level of DNA hemimethylation decreased from nonactivated 1-cell stage to 2-cell stage, then increased onward to blastocyst stage; moreover, the level of total methylation (including hemi- and methylation) decreased from nonactivated 1-cell stage to 4-cell stage, and then increased onward to blastocyst stage. This study indicated that incomplete DNA methylation could be a reason of low developmental ability and high loss rate of cloned embryos/fetuses in early pregnancy and low efficiency of somatic cell nuclear transfer.

REFERENCES

  1. Armstrong L., Lako M., Dean W. and Stojkovic M. (2006) Epigenetic modification is central to genome reprogramming in somatic cell nuclear transfer. Stem Cells, 24: 805-814.
  2. Aina R., Sgorbati S., Santagostino A., Labra M., Ghiani A. and Citterio S. (2004) Specific hypomethylation lf DNA is induced by heavy metals in white clover and industrial hemp. Physiol Plant, 121: 472-480.
  3. Beaujean N., Taylor J., Gardner J., Wilmut I., Meehan R. and Young L. (2004) Effect of limited DNA methylation reprogramming in the normal sheep embryo on somatic cell nuclear transfer. Bio. Reprod, 71: 185-193.
  4. Benhattar J. and Clément G. (2004) Methylation-sensitive single-strand conformation analysis: a rapid method to screen for and analyze DNA methylation. Methods Mol. Biol., 287: 181-193.
  5. Couldrey C. and Wells D.N. (2013) DNA methylation at a bovine alpha satellite I repeat CpG site during development following fertilization and somatic cell nuclear transfer. PLoS One, 8: e55153.
  6. de Montera B., Boulanger L., Taourit S., Renard J.P. and Eggen A. (2004) Genetic identity of clones and methods to explore DNA. Cloning Stem Cells, 6: 133-139.
  7. Han Y.M., Kang Y.K., Koo D.B. and Lee K.K. (2003) Nuclear reprogramming of cloned embryos produced in-vitro. Theriogenology, 59: 33-44.
  8. Herman J.G., Graff J.R., Myöhänen S., Nelkin B.D. and Baylin S.B. (1996) Methylation-specific PCR: a novel PCR assay for methylation status of CpG islands. Proc Natl Acad Sci USA, 93: 9821-9826.
  9. Kang Y.K., Koo D.B., Park J.S., Choi Y.H., Kim H.N., Chang W.K., Lee K.K. and Han Y.M. (2001) Typical demethylation events in cloned pig embryos. Clues on species-specific differences in epigenetic reprogramming of a cloned donor genome. J Biol Chem, 276: 39980–39984.
  10. Kang Y.K., Park J.S., Koo D.B., Choi Y.H., Kim S.U., Lee K.K. and Han Y.M. (2002) Limited demethylation leaves mosaictype methylation states in cloned bovine pre-implantation embryos. EMBO J, 21: 1092–1100.
  11. Karpinski P., Szmida E., Misiak B., Ramsey D., Leszczynski P., Bebenek M., Sedziak T., Grzebieniak Z., Jonkisz A., Lebioda A. and Sasiadek M.M. (2012) Assessment of three epigenotypes in colorectal cancer by combined bisulfite restriction analysis. Mol Carcinog, 51: 1003-1008.
  12. Lin L., Li Q., Zhang L., Zhao D., Dai Y. and Li N. (2008) Aberrant epigenetic changes and gene expression in cloned cattle dying around birth. BMC Dev Biol, 8: 14.
  13. Loi P., Ptak G., Barboni B., Fulka J.Jr., Cappai P. and Clinton M. (2001) Genetic rescue of an endangered mammal by cross-species nuclear transfer using post-mortem somatic cells. Nat Biotechnol, 19: 962-964.
  14. Ma L.B., Yang L., Hua S., Cao J.W., Li J.X. and Zhang Y. (2008a). Development in vitro and mitochondrial fate of interspecies cloned embryos. Reprod Domest Anim, 43: 279-285.
  15. Ma L.B., Yang L., Zhang Y., Cao J.W., Hua S. and Li J.X. (2008b) Quantitative analysis of mitochondrial RNA in goat- sheep cloned embryos. Mol Reprod Dev, 75: 33-39.
  16. Reik W., Dean W. and Walter J. (2001) Epigenetic reprogramming in mammalian development. Science, 293: 1089-1093.
  17. Xiong L.Z., Xu C.G., Saghai Maroof M.A. and Zhang Q. (1999) Patterns of cytosine methylation in an elite rice hybrid and its parental lines, detected by a methylation-sensitive amplification polymorphism technique. Mol Gen Genet, 261: 439-446.
Published In
Indian Journal of Animal Research
Article Metrics
Metrics Icon
0
Views
0
Citations
Reviewed By
Share Article
Download Article
In this Article
    Contact us
    Follow us

    volume 48 issue 3 (june 2014) : 221-226,   Doi: 10.5958/j.0976-0555.48.3.047

    THE LEVELS OF DNA METHYLATION OF SHEEP CLONED EMBRYOS IN DIFFERENT DEVELOPMENT STAGES

    L
    L.B. Ma*
    X
    X.Y. He
    1School of Mathematics, Physics and Biological Engineering, Inner Mongolia University of Science & Technology, Baotou, Inner Mongolia, China
    Cite article:- Ma* L.B., He X.Y. (2025). THE LEVELS OF DNA METHYLATION OF SHEEP CLONED EMBRYOS IN DIFFERENT DEVELOPMENT STAGES. Indian Journal of Animal Research. 48(3): 221-226. doi: 10.5958/j.0976-0555.48.3.047.

    ABSTRACT

    DNA methylation is a kind of epigenetic modification. During the early development of cloned embryos, parental genomic DNA undergoes active- and passive demethylation as well as low level of DNA methylation in cloned embryos at morula stage. In this study, sheep skin fibroblasts were used as nuclear donors, and sheep oocytes were used as nuclear recipient, sheep cloned embryos were constructed by the technology of somatic cell nuclear transfer, the development ability of cloned embryos was evaluated. Moreover, the dynamic change in DNA methylation level was also examined by the technology of methyl sensitive amplified polymorphism. The results showed that sheep cloned embryo had the ability to develop to blastocyst stage, and the level of DNA methylation decreased from nonactivated 1-cell stage to 4-cell stage, and then increased onward to blastocyst stage; while the level of DNA hemimethylation decreased from nonactivated 1-cell stage to 2-cell stage, then increased onward to blastocyst stage; moreover, the level of total methylation (including hemi- and methylation) decreased from nonactivated 1-cell stage to 4-cell stage, and then increased onward to blastocyst stage. This study indicated that incomplete DNA methylation could be a reason of low developmental ability and high loss rate of cloned embryos/fetuses in early pregnancy and low efficiency of somatic cell nuclear transfer.

    REFERENCES

    1. Armstrong L., Lako M., Dean W. and Stojkovic M. (2006) Epigenetic modification is central to genome reprogramming in somatic cell nuclear transfer. Stem Cells, 24: 805-814.
    2. Aina R., Sgorbati S., Santagostino A., Labra M., Ghiani A. and Citterio S. (2004) Specific hypomethylation lf DNA is induced by heavy metals in white clover and industrial hemp. Physiol Plant, 121: 472-480.
    3. Beaujean N., Taylor J., Gardner J., Wilmut I., Meehan R. and Young L. (2004) Effect of limited DNA methylation reprogramming in the normal sheep embryo on somatic cell nuclear transfer. Bio. Reprod, 71: 185-193.
    4. Benhattar J. and Clément G. (2004) Methylation-sensitive single-strand conformation analysis: a rapid method to screen for and analyze DNA methylation. Methods Mol. Biol., 287: 181-193.
    5. Couldrey C. and Wells D.N. (2013) DNA methylation at a bovine alpha satellite I repeat CpG site during development following fertilization and somatic cell nuclear transfer. PLoS One, 8: e55153.
    6. de Montera B., Boulanger L., Taourit S., Renard J.P. and Eggen A. (2004) Genetic identity of clones and methods to explore DNA. Cloning Stem Cells, 6: 133-139.
    7. Han Y.M., Kang Y.K., Koo D.B. and Lee K.K. (2003) Nuclear reprogramming of cloned embryos produced in-vitro. Theriogenology, 59: 33-44.
    8. Herman J.G., Graff J.R., Myöhänen S., Nelkin B.D. and Baylin S.B. (1996) Methylation-specific PCR: a novel PCR assay for methylation status of CpG islands. Proc Natl Acad Sci USA, 93: 9821-9826.
    9. Kang Y.K., Koo D.B., Park J.S., Choi Y.H., Kim H.N., Chang W.K., Lee K.K. and Han Y.M. (2001) Typical demethylation events in cloned pig embryos. Clues on species-specific differences in epigenetic reprogramming of a cloned donor genome. J Biol Chem, 276: 39980–39984.
    10. Kang Y.K., Park J.S., Koo D.B., Choi Y.H., Kim S.U., Lee K.K. and Han Y.M. (2002) Limited demethylation leaves mosaictype methylation states in cloned bovine pre-implantation embryos. EMBO J, 21: 1092–1100.
    11. Karpinski P., Szmida E., Misiak B., Ramsey D., Leszczynski P., Bebenek M., Sedziak T., Grzebieniak Z., Jonkisz A., Lebioda A. and Sasiadek M.M. (2012) Assessment of three epigenotypes in colorectal cancer by combined bisulfite restriction analysis. Mol Carcinog, 51: 1003-1008.
    12. Lin L., Li Q., Zhang L., Zhao D., Dai Y. and Li N. (2008) Aberrant epigenetic changes and gene expression in cloned cattle dying around birth. BMC Dev Biol, 8: 14.
    13. Loi P., Ptak G., Barboni B., Fulka J.Jr., Cappai P. and Clinton M. (2001) Genetic rescue of an endangered mammal by cross-species nuclear transfer using post-mortem somatic cells. Nat Biotechnol, 19: 962-964.
    14. Ma L.B., Yang L., Hua S., Cao J.W., Li J.X. and Zhang Y. (2008a). Development in vitro and mitochondrial fate of interspecies cloned embryos. Reprod Domest Anim, 43: 279-285.
    15. Ma L.B., Yang L., Zhang Y., Cao J.W., Hua S. and Li J.X. (2008b) Quantitative analysis of mitochondrial RNA in goat- sheep cloned embryos. Mol Reprod Dev, 75: 33-39.
    16. Reik W., Dean W. and Walter J. (2001) Epigenetic reprogramming in mammalian development. Science, 293: 1089-1093.
    17. Xiong L.Z., Xu C.G., Saghai Maroof M.A. and Zhang Q. (1999) Patterns of cytosine methylation in an elite rice hybrid and its parental lines, detected by a methylation-sensitive amplification polymorphism technique. Mol Gen Genet, 261: 439-446.
    In this Article
      Contact us
      Follow us
      Published In
      Indian Journal of Animal Research

      Editorial Board

      View all (0)