Dynamic changes in histone modification underlie the transition of gametic chromatin to embryonic chromatin. These changes are thought to be required for initiation of embryonic gene transcription and acquisition of pluripotency. Abnormalities in these transitions are likely to result in embryonic arrest at early preimplantation stages. Early embryonic mortality is a major cause of infertility in dairy cattle and humans. In bovine embryos, methylation of histone H3 lysine 27 is one of the epigenetic marks that are dynamically remodeled. Trimethylation of histone H3 lysine 27 is associated with silencing of transcription. We hypothesize that JMJD3, a histone H3 lysine 27 demethylases, is required for removal of methylation marks from H3K27, which in turn allows the initiation of embryonic gene transcription and normal embryo development. To test this hypothesis we will induce downregulation of JMJD3 in bovine oocytes and preimplantation embryos by siRNA injection.
Specific Aim 1 will determine whether maternally inherited JMJD3 is required for normal bovine embryonic development.
Specific Aim 2 will make use of chromatin immunoprecipitation to assess the role of JMJD3 on histone methylation reprogramming at the time of embryonic genome activation.
Specific aim 3 is designed to examine the role of JMJD3 on genome activation by analyzing the transcriptome of early embryos using RNA-Seq. Together;these experiments will expand our understanding of the basic mechanisms underlying embryonic development. This information will aid in developing diagnostic tools and interventions to treat infertility disorders in animals and people. Also, uncovering the nuclear reprogramming mechanisms elicited by the oocyte will help improve the success of somatic cell nuclear transfer for the generation of cloned animals for agricultural and biomedical applications and autologous stem cells for regenerative medicine applications.
A greater understanding of the biological mechanisms regulating epigenetic transformation during early embryonic development is critical to addressing problems of embryonic lethality observed in the agricultural production and human fertility fields. Results from this study will have direct impacts on the diagnosis and treatment of animal and human fertility disorders, while simultaneously bettering foundational knowledge that can be applied to nuclear reprogramming methodologies necessary for the generation of cloned animals and pluripotent stem cells.
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|Iqbal, Khursheed; Chitwood, James L; Meyers-Brown, Geraldine A et al. (2014) RNA-seq transcriptome profiling of equine inner cell mass and trophectoderm. Biol Reprod 90:61|
|Bakhtari, Azizollah; Ross, Pablo J (2014) DPPA3 prevents cytosine hydroxymethylation of the maternal pronucleus and is required for normal development in bovine embryos. Epigenetics 9:1271-9|
|Chitwood, James L; Rincon, Gonzalo; Kaiser, German G et al. (2013) RNA-seq analysis of single bovine blastocysts. BMC Genomics 14:350|
|Canovas, Sebastian; Cibelli, Jose B; Ross, Pablo J (2012) Jumonji domain-containing protein 3 regulates histone 3 lysine 27 methylation during bovine preimplantation development. Proc Natl Acad Sci U S A 109:2400-5|
|Bogliotti, Yanina S; Ross, Pablo J (2012) Mechanisms of histone H3 lysine 27 trimethylation remodeling during early mammalian development. Epigenetics 7:976-81|