| Cell cycle studies based upon quantitative image analysis. | |
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MedLine Citation:
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PMID: 18163464 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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When cell cycle studies are performed following cell cycle synchronization, it is possible that critical properties of an actively cycling cell will be overlooked. For this reason past studies have not revealed critical aspects of cell cycle control; such as how a cell determines when to exit the cell cycle, or how rapidly it should cycle. To address these challenging questions we have developed a procedure to quantitate fluorescent stains in a monolayer culture, where nuclear fluorescence and cell cycle history can be assessed with accuracy on a cell by cell basis. The cell cycle position of each cell can be determined by analyzing DNA and BrdU levels. The behavior of cells in a given cell cycle position can then be studied by quantitating up to two other stained markers. When the microinjection of siRNA, neutralizing antibodies, and expression plasmids are coupled with quantitative image analysis, these cell cycle studies can be conducted following alterations in the expression levels of selected cellular targets. With these techniques we have discovered critical aspects of cell cycle control; including how cyclin D1 levels vary through the cell cycle, the molecular mechanisms governing these changes, and the biological implications of changes in cyclin D1 concentration in various cell cycle stages. Our studies with cyclin D1, coupled with similar studies of p27Kip1, form the basis of an entirely new model of cell cycle control proposed here. This model explains how cell cycle progression is terminated, and how the length of the cell cycle is regulated. |
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Authors:
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Dennis W Stacey; Masahiro Hitomi |
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Publication Detail:
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Type: Journal Article; Research Support, N.I.H., Extramural; Review |
Journal Detail:
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Title: Cytometry. Part A : the journal of the International Society for Analytical Cytology Volume: 73 ISSN: 1552-4930 ISO Abbreviation: Cytometry A Publication Date: 2008 Apr |
Date Detail:
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Created Date: 2008-04-02 Completed Date: 2008-05-22 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 101235694 Medline TA: Cytometry A Country: United States |
Other Details:
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Languages: eng Pagination: 270-8 Citation Subset: IM |
Copyright Information:
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(c) 2007 International Society for Analytical Cytology. |
Affiliation:
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Department of Molecular Genetics, The Lerner Research Institute, The Cleveland Clinic Foundation, Cleveland, Ohio 44195, USA. staceyd@ccf.org |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Bromodeoxyuridine
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pharmacology Cell Cycle* Cell Proliferation Cyclin D1 / physiology* Cyclin-Dependent Kinase Inhibitor p27 / metabolism* DNA / metabolism Flow Cytometry / instrumentation, methods* Humans Microscopy, Fluorescence / methods Phosphorylation RNA, Small Interfering / metabolism |
| Grant Support | |
ID/Acronym/Agency:
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R01 52271//PHS HHS |
| Chemical | |
Reg. No./Substance:
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0/RNA, Small Interfering; 136601-57-5/Cyclin D1; 147604-94-2/Cyclin-Dependent Kinase Inhibitor p27; 59-14-3/Bromodeoxyuridine; 9007-49-2/DNA |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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