Document Detail

Cell-free extracts from mammalian oocytes partially induce nuclear reprogramming in somatic cells.
MedLine Citation:
PMID:  19164171     Owner:  NLM     Status:  MEDLINE    
Nuclear transfer has been regarded as the only reliable tool for studying nuclear reprogramming of mammalian somatic cells by oocytes. However, nuclear transfer is not well suited for biochemical analyses of the molecular mechanisms of reprogramming. A cell-free system from oocytes is an attractive alternative way to mimic reprogramming in vitro, since a large number of cells can be treated and analyzed. Nevertheless, a cell-free system using oocytes has not been developed in mammals. Here, cell extracts from porcine oocytes were prepared and their ability to induce nuclear reprogramming was evaluated. Extracts from metaphase II (MII) oocytes erased the machinery for regulating gene expression in reversibly permeabilized somatic cells. For example, the extracts caused histone deacetylation and the disappearance of TATA box-binding protein from the nuclei. However, MII-extract-treated cells did not show any obvious changes after cell culture. In contrast, extracts from germinal vesicle (GV) oocytes activated pluripotent marker genes, especially NANOG, and induced partial dedifferentiation after cell culture. The activation of pluripotent marker genes by GV extracts was associated with histone acetylation that was induced during extract treatment. These results indicate that GV- and MII-oocyte extracts have different roles on nuclear reprogramming. Furthermore, both oocyte extracts induced site-specific demethylation in the upstream region of NANOG. These results indicate that cell-free extracts derived from GV- and MII-oocytes could be useful for studying the mechanisms involved in nuclear reprogramming.
Kei Miyamoto; Tomoyuki Tsukiyama; Yang Yang; Ning Li; Naojiro Minami; Masayasu Yamada; Hiroshi Imai
Publication Detail:
Type:  In Vitro; Journal Article; Research Support, Non-U.S. Gov't     Date:  2009-01-21
Journal Detail:
Title:  Biology of reproduction     Volume:  80     ISSN:  0006-3363     ISO Abbreviation:  Biol. Reprod.     Publication Date:  2009 May 
Date Detail:
Created Date:  2009-04-29     Completed Date:  2009-06-19     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0207224     Medline TA:  Biol Reprod     Country:  United States    
Other Details:
Languages:  eng     Pagination:  935-43     Citation Subset:  IM    
Laboratory of Reproductive Biology, Graduate School of Agriculture, Kyoto University, Kitashirakawa, Kyoto, Japan.
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MeSH Terms
Acetylation / drug effects
Cell Differentiation / drug effects
Cell Extracts / pharmacology
Cell Nucleus / drug effects*,  metabolism*
Cell-Free System
Cells, Cultured
DNA Methylation / drug effects
Genes, Homeobox / drug effects
Histones / metabolism
Nuclear Reprogramming / drug effects*,  genetics,  physiology
Oocytes / metabolism*
Sus scrofa
Transcriptional Activation / drug effects
Reg. No./Substance:
0/Cell Extracts; 0/Histones

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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