73 ABNORMAL REPROGRAMMING OF HISTONE ACETYLATION IN CLONED BOVINE EMBRYOS
G. Wee A , D.-B. Koo A , M.-J. Kang B , S.J. Moon B , K.-K. Lee A and Y.-M. Han AA Laboratory of Development and Differentiation, Korea Research Institute of Bioscience and Biotechnology, Daejeon, Korea
B Department of Animal Science, Chonnam National University, Gwangju, Korea. Email: ymhan@kribb.re.kr
Reproduction, Fertility and Development 17(2) 186-187 https://doi.org/10.1071/RDv17n2Ab73
Submitted: 1 August 2004 Accepted: 1 October 2004 Published: 1 January 2005
Abstract
Histone acetylation plays an important role in the chromatin structure prior to zygotic gene expression during early embryonic development. Successful animal clones indicate that differentiated somatic nuclei must be reprogrammed to some extent during pre-implantation development. However, the molecular mechanisms regarding epigenetic reprogramming of somatic nuclei in the early-stage embryos are poorly understood. To test this, the patterns of hyperacetylated histone H4 lysine 5 (AcH4K5) in the nuclear-transferred (NT) embryos were monitored, comparing in vitro fertilized (IVF) embryos and Trichostatin A (TSA)-NT embryos with TSA-treated cells. The intensity signals of AcH4K5 were observed in early-stage embryos and somatic cells (bovine ear skin fibroblasts composed of about 80% at G0/G1 stage) by immunofluorescence analysis with anti-AcH4K5 using image the analyzer system, SigmaScan-pro V5.01 (SPSS, Inc., Chicago, IL, USA). Our data were analyzed by analysis of variance (ANOVA) using an SAS package (SAS Institute, Inc., Cary, NC, USA). Somatic cells were exposed to TSA (1 μM for 60 h), a specific inhibitor of histone deacetylase (HDAC), to induce hyperacetylation prior to somatic cell nuclear transfer. Signal intensity for AcH4K5 in TSA-treated cells (n = 80) was significantly increased (P < 0.05), which was approximately double compared to that of normal cells (n = 80). In normal cells, histone H4 acetylation was profoundly reduced from the pro-metaphase to the early telophase and then reappeared at the late telophase. Acetylation signals of TSA-treated cells gradually increased to the early anaphase, abruptly decreased at the late anaphase and the early telophase, and recovered during late telophase. During early embryonic development (1 cell to 8 cell stage), NT embryos (n = 8) were hypoacetylated at the metaphase, whereas IVF (n = 10) and TSA-NT embryos (n = 8) were hyperacetylated. Our findings demonstrate that aberrant epigenetic reprogramming of histone modification occurs as early as the pronuclear stage in cloned embryos.
