Henderson, S., Allsopp, R., Spector, D., Wang, S. S., Harley, C. (July 1996) In situ analysis of changes in telomere size during replicative aging and cell transformation. Journal of Cell Biology, 134 (1). pp. 1-12. ISSN 0021-9525 (Print)0021-9525
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Abstract
Telomeres have been shown to gradually shorten during replicative aging in human somatic cells by Southern analysis. This study examines telomere shortening at the single cell level by fluorescence in situ hybridization (FISH). FISH and confocal microscopy of interphase human diploid fibroblasts (HDFs) demonstrate that telomeres are distributed throughout the nucleus with an interchromosomal heterogeneity in size. Analysis of HDFs at increasing population doubling levels shows a gradual increase in spot size, intensity, and detectability of telomeric signal. FISH of metaphase chromosomes prepared from young and old HDFs shows a heterogeneity in detection frequency for telomeres on chromosomes 1, 9, 15, and Y. The interchromosomal distribution of detection frequencies was similar for cells at early and late passage. The telomeric detection frequency for metaphase chromosomes also decreased with age. These observations suggest that telomeres shorten at similar rates in normal human somatic cels. T-antigen transformed HDFs near crisis contained telomere signals that were low compared to nontransformed HDFs. A large intracellular heterogeneity in telomere lengths was detected in two telomerase-negative cell lines compared to normal somatic cells and the telomerase-positive 293 cell line. Many telomerase-negative immortal cells had telomeric signals stronger than those in young HDFs, suggesting a different mechanism for telomere length regulation in telomerase-negative immortal cells. These studies provide an in situ demonstration of interchromosomal heterogeneity in telomere lengths. Furthermore, FISH is a reliable and sensitive method for detecting changes in telomere size at the single cell level.
| Item Type: | Paper |
|---|---|
| Uncontrolled Keywords: | *Cell Aging Cell Division *Cell Transformation, Viral Cells, Cultured Humans In Situ Hybridization, Fluorescence Simian virus 40 Telomere/*ultrastructure |
| Subjects: | bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > DNA replication bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > telomeres |
| CSHL Authors: | |
| Communities: | CSHL labs > Spector lab |
| Depositing User: | Matt Covey |
| Date: | July 1996 |
| Date Deposited: | 26 Mar 2014 20:00 |
| Last Modified: | 23 Jan 2015 20:10 |
| PMCID: | PMC2120915 |
| Related URLs: | |
| URI: | https://repository.cshl.edu/id/eprint/29705 |
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