Document Detail


Beta-galactosidase histochemistry and telomere loss in senescent retinal pigment epithelial cells.
MedLine Citation:
PMID:  9888444     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
H Matsunaga; J T Handa; A Aotaki-Keen; S W Sherwood; M D West; L M Hjelmeland
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Investigative ophthalmology & visual science     Volume:  40     ISSN:  0146-0404     ISO Abbreviation:  Invest. Ophthalmol. Vis. Sci.     Publication Date:  1999 Jan 
Date Detail:
Created Date:  1999-01-20     Completed Date:  1999-01-20     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  7703701     Medline TA:  Invest Ophthalmol Vis Sci     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  197-202     Citation Subset:  IM    
Affiliation:
Department of Ophthalmology, University of California Davis, USA.
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MeSH Terms
Descriptor/Qualifier:
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:
EY00344/EY/NEI NIH HHS; EY06473/EY/NEI NIH HHS
Chemical
Reg. No./Substance:
59-14-3/Bromodeoxyuridine; 9007-49-2/DNA; EC 3.2.1.23/beta-Galactosidase
Comments/Corrections
Comment In:
Invest Ophthalmol Vis Sci. 1999 Jan;40(1):1-2   [PMID:  9888419 ]

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


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