| Cell-free extracts from mammalian oocytes partially induce nuclear reprogramming in somatic cells. | |
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MedLine Citation:
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PMID: 19164171 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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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. |
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Authors:
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Kei Miyamoto; Tomoyuki Tsukiyama; Yang Yang; Ning Li; Naojiro Minami; Masayasu Yamada; Hiroshi Imai |
Publication Detail:
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Type: In Vitro; Journal Article; Research Support, Non-U.S. Gov't Date: 2009-01-21 |
Journal Detail:
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Title: Biology of reproduction Volume: 80 ISSN: 0006-3363 ISO Abbreviation: Biol. Reprod. Publication Date: 2009 May |
Date Detail:
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Created Date: 2009-04-29 Completed Date: 2009-06-19 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 0207224 Medline TA: Biol Reprod Country: United States |
Other Details:
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Languages: eng Pagination: 935-43 Citation Subset: IM |
Affiliation:
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Laboratory of Reproductive Biology, Graduate School of Agriculture, Kyoto University, Kitashirakawa, Kyoto, Japan. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Acetylation
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drug effects Animals Cell Differentiation / drug effects Cell Extracts / pharmacology Cell Nucleus / drug effects*, metabolism* Cell-Free System Cells, Cultured DNA Methylation / drug effects Female Genes, Homeobox / drug effects Histones / metabolism Nuclear Reprogramming / drug effects*, genetics, physiology Oocytes / metabolism* Sus scrofa Transcriptional Activation / drug effects |
| Chemical | |
Reg. No./Substance:
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0/Cell Extracts; 0/Histones |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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