Effects of histone acetylation status on the early development of in vitro porcine transgenic cloned embryos.
The purpose of this study was to investigate the effects of the histone deacetylase (HDAC) inhibitor trichostatin A (TSA) on transgene expression and development of porcine transgenic cloned embryos, specifically focusing on effects derived from TSA-treated donor cells or TSA-treated reconstructed embryos. The results showed that TSA treatment on reconstructed embryos modified the acetylation status, which significantly improved the development of porcine somatic cell nuclear transfer (SCNT) embryos in vitro, but not donor cells. Furthermore, the treatment of reconstructed embryos with TSA enhanced expression of the pluripotency-related gene POU5F1 and stimulated expression of the anti-apoptotic gene BCL-2. Enhanced green fluorescent protein (EGFP) mRNA expression of every group dropped drastically from donor cells to blastocysts. Interestingly, TSA is likely to prevent a decline in EGFP expression in nuclear reprogramming of porcine SCNT embryos. However DNA hypomethylation induced by modified histone acetylation of donor cells treated with TSA was significantly more effective in increasing EGFP expression in SCNT blastocysts. In conclusion, the acetylation status of both donor cells and reconstructed embryos modified by TSA treatment increased transgene expression and improved nuclear reprogramming and the developmental potential of porcine transgenic SCNT embryos.
1 College of Veterinary Medicine, Nanjing Agricultural University , Jiangsu, 210095, China .
Biping
B
Luo
Shiqiang
S
Ju
Caroline W
CW
Muneri
Rong
R
Rui
D000107:Acetylation; D000818:Animals; D030801:Animals, Genetically Modified; D001755:Blastocyst / Q000378:metabolism*; D019976:Cloning, Organism; D019175:DNA Methylation; D046149:Embryo Culture Techniques; D047108:Embryonic Development / Q000187:drug effects*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D056572:Histone Deacetylase Inhibitors; D006657:Histones / Q000378:metabolism*; D006877:Hydroxamic Acids / Q000494:pharmacology*; D053652:Nuclear Transfer Techniques / Q000662:veterinary*; D050814:Octamer Transcription Factor-3 / Q000378:metabolism; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D013552:Swine
D056572:Histone Deacetylase Inhibitors; D006657:Histones; D006877:Hydroxamic Acids; D050814:Octamer Transcription Factor-3; D019253:Proto-Oncogene Proteins c-bcl-2; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; C012589:trichostatin A