Document Detail


Oocyte extracts for the study of meiotic M-M transition.
MedLine Citation:
PMID:  16739743     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
In meiotic cell cycles, meiosis I (MI) is followed by meiosis II (MII) without an intervening S phase, whereas in mitotic cell cycles, an S phase necessarily alternates with an M phase. For the study of mitotic cell cycles, extracts prepared from unfertilized and parthenogenetically activated Xenopus eggs have been very useful as they can perform the progression of mitotic cycles in vitro. To establish a cell-free system to study the regulatory mechanisms of meiotic transition from MI to MII, extracts have been prepared from maturing Xenopus oocytes isolated from ovaries, stimulated with progesterone to induce the resumption of meiosis, and arrested at meiotic metaphase I by cold treatment. In oocyte extracts, the activity of cyclin B-Cdc2 complexes, the M phase inducer, fluctuates in the same manner as it does in maturing oocytes during the MI to MII transition period. By the use of oocyte extracts, it has been found that incomplete inactivation of Cdc2 at the end of MI is required for meiotic M-M transition. The meiotic extract should provide a useful tool to elucidate molecular mechanisms of meiotic M to M transition, including a role of Mos/mitogen-activated protein kinase cascade in the suppression of S phase entry after MI exit. In this chapter, we describe methods for the preparation and the uses of meiotic extracts. As a comparison, we also include a protocol for the preparation of mitotic extracts.
Authors:
Keita Ohsumi; Tomomi M Yamamoto; Mari Iwabuchi
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Publication Detail:
Type:  Journal Article; Review    
Journal Detail:
Title:  Methods in molecular biology (Clifton, N.J.)     Volume:  322     ISSN:  1064-3745     ISO Abbreviation:  Methods Mol. Biol.     Publication Date:  2006  
Date Detail:
Created Date:  2006-06-02     Completed Date:  2006-06-30     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9214969     Medline TA:  Methods Mol Biol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  445-58     Citation Subset:  IM    
Affiliation:
Laboratory of Cell and Developmental Biology, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Yokohama, Japan.
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MeSH Terms
Descriptor/Qualifier:
Animals
Cell Extracts*
Cell-Free System
Female
Male
Meiosis / physiology*
Oocytes / cytology*,  physiology*
Spermatozoa / physiology
Xenopus laevis
Chemical
Reg. No./Substance:
0/Cell Extracts

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


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