| Beta-galactosidase histochemistry and telomere loss in senescent retinal pigment epithelial cells. | |
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MedLine Citation:
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PMID: 9888444 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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PURPOSE: To investigate the relation of senescence-related beta-galactosidase activity and telomere shortening to replicative senescence in cultured human retinal pigment epithelial (RPE) cells. METHODS: A human RPE cell line was serially passaged until 80% of cells were nondividing in a 72-hour 5-bromo-2'-deoxyuridine (BrdU) labeling study. Early- and late-passage cells were double-stained for BrdU and senescence-related beta-galactosidase activity (pH 6). The average chromosomal telomere length at several population doublings was estimated by Southern blot analysis after double digestion of DNA with RsaI and HinfI and using a telomere-specific probe. RESULTS: BrdU-beta-galactosidase double-staining revealed an inverse correlation between the number of BrdU-labeled nuclei and beta-galactosidase-labeled cells as a function of population doubling level (PDL). At PDL 58, only 20% of all cells labeled for BrdU, whereas 57% stained for beta-galactosidase. The mean terminal restriction fragment length (TRF) was reduced from 10 kb in early (PDL 12) cultures to 4 kb in late (PDL 57) cultures. CONCLUSIONS: Senescence-related beta-galactosidase activity and mean TRF length may prove useful in studying the senescence of RPE cells in vitro. These techniques may be valuable in determining senescence of the retinal pigment epithelium in vivo, where senescent RPE cells could be involved in the development of age-related maculopathy and age-related macular degeneration. |
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Authors:
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H Matsunaga; J T Handa; A Aotaki-Keen; S W Sherwood; M D West; L M Hjelmeland |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S. |
Journal Detail:
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Title: Investigative ophthalmology & visual science Volume: 40 ISSN: 0146-0404 ISO Abbreviation: Invest. Ophthalmol. Vis. Sci. Publication Date: 1999 Jan |
Date Detail:
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Created Date: 1999-01-20 Completed Date: 1999-01-20 Revised Date: 2007-11-14 |
Medline Journal Info:
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Nlm Unique ID: 7703701 Medline TA: Invest Ophthalmol Vis Sci Country: UNITED STATES |
Other Details:
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Languages: eng Pagination: 197-202 Citation Subset: IM |
Affiliation:
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Department of Ophthalmology, University of California Davis, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Blotting, Southern Bromodeoxyuridine / metabolism Cell Aging* / physiology Cell Division Cell Line Cells, Cultured DNA / analysis DNA Replication Histocytochemistry Humans Infant Pigment Epithelium of Eye / cytology, enzymology* Telomere / metabolism* beta-Galactosidase / metabolism* |
| Grant Support | |
ID/Acronym/Agency:
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EY00344/EY/NEI NIH HHS; EY06473/EY/NEI NIH HHS |
| Chemical | |
Reg. No./Substance:
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59-14-3/Bromodeoxyuridine; 9007-49-2/DNA; EC 3.2.1.23/beta-Galactosidase |
| Comments/Corrections | |
Comment In:
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Invest Ophthalmol Vis Sci. 1999 Jan;40(1):1-2
[PMID:
9888419
]
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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