TSA对克隆胚胎染色质结构相关基因,DNA甲基化与发育相关基因表达的影响

大更丰富

CLONING AND STEM CELLSVolume 10, Number 1, 2008© Mary Ann Liebert, Inc.DOI: 10.1089/clo.2007.0066

The Effects of Trichostatin A on mRNA Expression

of Chromatin Structure-, DNA Methylation-,

and Development-Related Genes in

Cloned Mouse Blastocysts

XIANGPING LI,* YOKO KATO,* YUTA TSUJI, and YUKIO TSUNODA

ABSTRACT

Trichostatin A (TSA) is the most potent histone deacetylase (HDAC) inhibitor known. Wepreviously reported that treatment of mouse somatic cell nuclear-transferred (SCNT) oocyteswith TSA significantly increased the blastocyst rate, blastocyst cell number, and full-term de-velopment. How TSA enhances the epigenetic remodeling ability of somatic nuclei and theexpression of development-related genes, however, is not known. In the present study, wecompared the expression patterns of nine genes involved in chromatin structure and DNAmethylation, and seven development-related genes in blastocysts developed from SCNToocytes treated with and without TSA, and in blastocysts developed in vivoand in vitrous-ing real-time reverse transcription-polymerase chain reaction. In vivo-recovered blastocystsand blastocysts developed from TSA-treated SCNT oocytes exhibited similar expression pat-terns for Hdac1, 2, and 3, CBP, PCAF, and Dnmt3b genes compared with in vitro-developedblastocysts and blastocysts developed from SCNT oocytes without TSA treatment. There weresignificantly lower expression levels of Hdac1 and Hdac2 transcripts in TSA-treated and invivo-recovered blastocysts than in TSA-untreated and in vitro-developed blastocysts. Thefinding that TSA treatment of SCNT oocytes significantly upregulated Sox2 and cMyc tran-scripts in blastocysts indicated that both transcripts are TSA-responsive genes. Thus, TSAtreatment of mouse SCNT oocytes decreased the expression of chromatin structure- and DNAmethylation-related genes, and increased the expression of Sox2 and cMyc genes in blasto-cysts. Such modifications might be a reason for the high developmental potential of mouseSCNT oocytes treated with TSA.

cessfully cloned from adult somatic cells. Al-though mice were cloned from somatic cells in

HEFIRSTSUCCESSFULnuclear transplantation 1998 (Wakayama et al., 1998), production effi-in mammals was performed in mice (Mc-ciency of cloned mice is rather lower comparedGrath and Solter, 1983). In 1997, sheep (Wilmutwith bovine (Tsunoda and Kato, 2002). Moreover,et al., 1997) were the first mammalian species suc-cloned mice have a high rate of abnormalities,

INTRODUCTION

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Laboratory of Animal Reproduction, College of Agriculture, Kinki University, Nara, Japan.*These authors contributed equally to this work.

Present address: Animal Reproduction Institute, Guangxi University. Daxue Road 100, Nanning 530005, P.R. China.

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