Document Detail

Trichostatin A improved epigenetic modifications of transfected cells but did not improve subsequent cloned embryo development.
MedLine Citation:
PMID:  18855246     Owner:  NLM     Status:  MEDLINE    
Reprogramming impairment of DNA methylation may be partly responsible for the low efficiency in somatic cell nuclear transfer. In this study, bovine fibroblast cells were transfected with enhancer green fluorescence protein (eGFP), and then treated with a histone-deacetylase inhibitor, trichostatin A (TSA). The results showed that the effect of TSA on transfected cells was dose dependent. When the TSA concentration was over 5 ng/ml, cell proliferation was significantly inhibited. The majority of the cells died when TSA reached 100 ng/ml (P < 0.01). The number of cells in the S phase was significantly decreased in the 5- to 50-ng/ml TSA-treated groups, while the majority of the cells were at the G0/G1 phases. The number of eGFP-expressed cells were approximately twofold higher in 25-ng/ml (30.5%) and 50-ng/ml (29.5%) TSA groups than the control (15.0%). Reduced DNA methylation and improved histone acetylation were observed when the cells were treated with 10 to 50 ng/ml of TSA. Transfer of the TSA-treated cells to enucleated recipient oocytes resulted in similar cleavage rates among the experimental groups and the control. Cells treated with 50 ng/ml of TSA resulted in significantly lower blastocyst development (9.9%) than the other experimental and the control groups (around 20%). Analysis of the putative blastocysts showed that 86.7% of the embryos derived from TSA-treated cells were eGFP positive, which was higher than that from untreated cells (68.8%). In conclusion, treatment of transfected cells with TSA decreased the genome DNA methylation level, increased histone acetylation, and eGFP gene expression was activated. Donor cells with reduced DNA methylation did not improve subsequent cloned embryo development; however, transgene expression was improved in cloned embryos.
Xia Wu; Yan Li; Guang-Peng Li; Dongshan Yang; Yongli Yue; Lingling Wang; Kehan Li; Penghui Xin; Shorgan Bou; Haiquan Yu
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Animal biotechnology     Volume:  19     ISSN:  1532-2378     ISO Abbreviation:  Anim. Biotechnol.     Publication Date:  2008  
Date Detail:
Created Date:  2008-10-15     Completed Date:  2008-12-24     Revised Date:  2009-11-19    
Medline Journal Info:
Nlm Unique ID:  9011409     Medline TA:  Anim Biotechnol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  211-24     Citation Subset:  IM    
The Key Laboratory of Mammalian Reproductive Biology and Biotechnology of the Ministry of Education, Inner Mongolia University, Hohhot, China.
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MeSH Terms
Acetylation / drug effects
Cattle / embryology*,  genetics,  metabolism
Cloning, Organism
DNA Methylation / drug effects
Embryonic Development / drug effects*,  genetics
Enzyme Inhibitors / pharmacology
Epigenesis, Genetic / drug effects
Fibroblasts / drug effects,  physiology
Green Fluorescent Proteins / biosynthesis,  genetics
Histone Deacetylase Inhibitors
Histones / metabolism
Hydroxamic Acids / pharmacology*
Nuclear Transfer Techniques / veterinary*
Transfection / veterinary
Reg. No./Substance:
0/Enzyme Inhibitors; 0/Histone Deacetylase Inhibitors; 0/Histones; 0/Hydroxamic Acids; 0/enhanced green fluorescent protein; 147336-22-9/Green Fluorescent Proteins; 58880-19-6/trichostatin A

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