Home LiteratureArticle Details
PMID: 8698806 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

In situ analysis of changes in telomere size during replicative aging and cell transformation.

The Journal of cell biology ·Vol. 134 ·No. 1 ·1996-07-00 ·Pages 1-12

Henderson S, Allsopp R, Spector D, Wang SS, Harley C

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 decrease 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.

MeSH Terms
Cell Division Cell Transformation, Viral Cells, Cultured Cellular Senescence Humans In Situ Hybridization, Fluorescence Simian virus 40 Telomere/ultrastructure
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Henderson S
Cold Spring Harbor Laboratory, Cold Spring Harbor, New York 11724, USA.
Allsopp R
Spector D
Wang S S
Harley C
References (67)
67 references, click to expand
  1. Cellular senescence revisited: a review.
    Mech Ageing Dev. 1987 Mar;38(1):1-48 PMID: 2439851
  2. Characterization and organization of DNA sequences adjacent to the human telomere associated repeat (TTAGGG)n.
    Nucleic Acids Res. 1990 Jun 11;18(11):3353-61 PMID: 2356126
  3. The Stability of Broken Ends of Chromosomes in Zea Mays.
    Genetics. 1941 Mar;26(2):234-82 PMID: 17247004
  4. Origin of concatemeric T7 DNA.
    Nat New Biol. 1972 Oct 18;239(94):197-201 PMID: 4507727
  5. Telomeres shorten during ageing of human fibroblasts.
    Nature. 1990 May 31;345(6274):458-60 PMID: 2342578
  6. Evidence for a mitotic clock in human hematopoietic stem cells: loss of telomeric DNA with age.
    Proc Natl Acad Sci U S A. 1994 Oct 11;91(21):9857-60 PMID: 7937905
  7. Time, telomeres and tumours: is cellular senescence more than an anticancer mechanism?
    Trends Cell Biol. 1995 Aug;5(8):293-7 PMID: 14732086
  8. Reproducible compartmentalization of individual chromosome domains in human CNS cells revealed by in situ hybridization and three-dimensional reconstruction.
    Chromosoma. 1988;96(6):397-410 PMID: 3219911
  9. Correlating telomerase activity levels with human neuroblastoma outcomes.
    Nat Med. 1995 Mar;1(3):249-55 PMID: 7585042
  10. Human naive and memory T lymphocytes differ in telomeric length and replicative potential.
    Proc Natl Acad Sci U S A. 1995 Nov 21;92(24):11091-4 PMID: 7479943
  11. A simple method of reducing the fading of immunofluorescence during microscopy.
    J Immunol Methods. 1981;43(3):349-50 PMID: 7019347
  12. DNA damage triggers a prolonged p53-dependent G1 arrest and long-term induction of Cip1 in normal human fibroblasts.
    Genes Dev. 1994 Nov 1;8(21):2540-51 PMID: 7958916
  13. Telomeres and telomerase in aging and cancer.
    Curr Opin Genet Dev. 1995 Apr;5(2):249-55 PMID: 7613096
  14. Unusual chromatin in human telomeres.
    Mol Cell Biol. 1994 Sep;14(9):5777-85 PMID: 8065312
  15. Telomere length predicts replicative capacity of human fibroblasts.
    Proc Natl Acad Sci U S A. 1992 Nov 1;89(21):10114-8 PMID: 1438199
  16. Human telomeres are attached to the nuclear matrix.
    EMBO J. 1992 Feb;11(2):717-24 PMID: 1537344
  17. Evidence for a critical telomere length in senescent human fibroblasts.
    Exp Cell Res. 1995 Jul;219(1):130-6 PMID: 7628529
  18. Telomerase activity in normal and malignant hematopoietic cells.
    Proc Natl Acad Sci U S A. 1995 Sep 26;92(20):9082-6 PMID: 7568077
  19. Specific chromosome aberrations in senescent fibroblast cell lines derived from human embryos.
    Am J Hum Genet. 1976 Sep;28(5):465-73 PMID: 984042
  20. Telomere end-replication problem and cell aging.
    J Mol Biol. 1992 Jun 20;225(4):951-60 PMID: 1613801
  21. Localization of ribosomal and telomeric DNA sequences in intact plant nuclei by in-situ hybridization and three-dimensional optical microscopy.
    J Microsc. 1990 Jan;157(Pt 1):83-9 PMID: 2299663
  22. Interphase chromosomes of Friend-S cells are attached to the matrix structures through the centromeric/telomeric regions.
    DNA Cell Biol. 1994 Sep;13(9):941-51 PMID: 7522463
  23. Telomere-related sequences at interstitial sites in the human genome.
    Genomics. 1990 Dec;8(4):699-704 PMID: 2276741
  24. Replicative senescence: the human fibroblast comes of age.
    Science. 1990 Sep 7;249(4973):1129-33 PMID: 2204114
  25. Telomere dynamics in an immortal human cell line.
    EMBO J. 1994 Oct 17;13(20):4953-62 PMID: 7957062
  26. A theory of marginotomy. The incomplete copying of template margin in enzymic synthesis of polynucleotides and biological significance of the phenomenon.
    J Theor Biol. 1973 Sep 14;41(1):181-90 PMID: 4754905
  27. Aging in vitro. Growth of cultured cells from the Galapagos tortoise.
    Exp Cell Res. 1974 Feb;83(2):297-302 PMID: 4816902
  28. Telomere G-strand structure and function analyzed by chemical protection, base analogue substitution, and utilization by telomerase in vitro.
    Biochemistry. 1990 Jan 23;29(3):732-7 PMID: 2337592
  29. Structure and function of telomeres.
    Nature. 1991 Apr 18;350(6319):569-73 PMID: 1708110
  30. Doubling potential, calendar time, and senescence of human diploid cells in culture.
    Exp Cell Res. 1973 Mar 15;77(1):356-60 PMID: 4570356
  31. Structure and polymorphism of human telomere-associated DNA.
    Cell. 1990 Oct 5;63(1):119-32 PMID: 2208276
  32. Loss of telomeric DNA during aging of normal and trisomy 21 human lymphocytes.
    Am J Hum Genet. 1993 Apr;52(4):661-7 PMID: 8460632
  33. Mapping of human chromosome Xq28 by two-color fluorescence in situ hybridization of DNA sequences to interphase cell nuclei.
    Am J Hum Genet. 1991 Jan;48(1):1-15 PMID: 1985451
  34. Specific association of human telomerase activity with immortal cells and cancer.
    Science. 1994 Dec 23;266(5193):2011-5 PMID: 7605428
  35. Telomere shortening associated with chromosome instability is arrested in immortal cells which express telomerase activity.
    EMBO J. 1992 May;11(5):1921-9 PMID: 1582420
  36. Cell cycle dependent chromosomal movement in pre-mitotic human T-lymphocyte nuclei.
    Chromosoma. 1992 Aug;101(9):557-65 PMID: 1521500
  37. Replicative life-span of cultivated human cells. Effects of donor's age, tissue, and genotype.
    Lab Invest. 1970 Jul;23(1):86-92 PMID: 5431223
  38. Localization of centromeric satellite and telomeric DNA sequences in dorsal root ganglion neurons, in vitro.
    J Cell Sci. 1991 Sep;100 ( Pt 1):219-26 PMID: 1795027
  39. A mutant with a defect in telomere elongation leads to senescence in yeast.
    Cell. 1989 May 19;57(4):633-43 PMID: 2655926
  40. The RNA component of human telomerase.
    Science. 1995 Sep 1;269(5228):1236-41 PMID: 7544491
  41. Modification of DAPI banding on human chromosomes by prestaining with a DNA-binding oligopeptide antibiotic, distamycin A.
    Exp Cell Res. 1978 Feb;111(2):327-32 PMID: 75107
  42. Loss of a yeast telomere: arrest, recovery, and chromosome loss.
    Cell. 1993 Nov 19;75(4):729-39 PMID: 8242745
  43. In vivo loss of telomeric repeats with age in humans.
    Mutat Res. 1991 Jan;256(1):45-8 PMID: 1944386
  44. A telomeric sequence in the RNA of Tetrahymena telomerase required for telomere repeat synthesis.
    Nature. 1989 Jan 26;337(6205):331-7 PMID: 2463488
  45. Analysis of genes and chromosomes by nonisotopic in situ hybridization.
    Genet Anal Tech Appl. 1991 Feb;8(1):24-35 PMID: 2043382
  46. Defining cellular senescence in IMR-90 cells: a flow cytometric analysis.
    Proc Natl Acad Sci U S A. 1988 Dec;85(23):9086-90 PMID: 3194411
  47. Differential expression of telomerase activity in hematopoietic progenitors from adult human bone marrow.
    Stem Cells. 1996 Mar;14(2):239-48 PMID: 8991544
  48. Well-defined genome architecture in the human sperm nucleus.
    Chromosoma. 1995 May;103(9):577-90 PMID: 7587580
  49. Enzymatic synthesis of biotin-labeled polynucleotides: novel nucleic acid affinity probes.
    Proc Natl Acad Sci U S A. 1981 Nov;78(11):6633-7 PMID: 6273878
  50. Identification of a specific telomere terminal transferase activity in Tetrahymena extracts.
    Cell. 1985 Dec;43(2 Pt 1):405-13 PMID: 3907856
  51. Telomerase activity in normal leukocytes and in hematologic malignancies.
    Blood. 1995 May 1;85(9):2315-20 PMID: 7727765
  52. Acquisition of telomere repeat sequences by transfected DNA integrated at the site of a chromosome break.
    Mol Cell Biol. 1993 Feb;13(2):977-83 PMID: 8423817
  53. [Principle of marginotomy in template synthesis of polynucleotides].
    Dokl Akad Nauk SSSR. 1971;201(6):1496-9 PMID: 5158754
  54. A highly conserved repetitive DNA sequence, (TTAGGG)n, present at the telomeres of human chromosomes.
    Proc Natl Acad Sci U S A. 1988 Sep;85(18):6622-6 PMID: 3413114
  55. Structure and variability of human chromosome ends.
    Mol Cell Biol. 1990 Feb;10(2):518-27 PMID: 2300052
  56. Human telomeres contain at least three types of G-rich repeat distributed non-randomly.
    Nucleic Acids Res. 1989 Jun 26;17(12):4611-27 PMID: 2664709
  57. Telomere loss: mitotic clock or genetic time bomb?
    Mutat Res. 1991 Mar-Nov;256(2-6):271-82 PMID: 1722017
  58. Cell cycle-dependent distribution of telomeres, centromeres, and chromosome-specific subsatellite domains in the interphase nucleus of mouse lymphocytes.
    Exp Cell Res. 1993 Mar;205(1):142-51 PMID: 8453988
  59. Telomere elongation in immortal human cells without detectable telomerase activity.
    EMBO J. 1995 Sep 1;14(17):4240-8 PMID: 7556065
  60. Telomere reduction in human colorectal carcinoma and with ageing.
    Nature. 1990 Aug 30;346(6287):866-8 PMID: 2392154
  61. Telomeres and the functional architecture of the nucleus.
    Trends Cell Biol. 1993 Apr;3(4):128-34 PMID: 14731767
  62. Staining of human telomeres with primed in situ labeling (PRINS).
    Cytogenet Cell Genet. 1995;68(1-2):115-8 PMID: 7525160
  63. Conservation of the human telomere sequence (TTAGGG)n among vertebrates.
    Proc Natl Acad Sci U S A. 1989 Sep;86(18):7049-53 PMID: 2780561
  64. Telomere length and replicative aging in human vascular tissues.
    Proc Natl Acad Sci U S A. 1995 Nov 21;92(24):11190-4 PMID: 7479963
  65. Telomerase activity in human ovarian carcinoma.
    Proc Natl Acad Sci U S A. 1994 Apr 12;91(8):2900-4 PMID: 8159676
  66. Stabilization of short telomeres and telomerase activity accompany immortalization of Epstein-Barr virus-transformed human B lymphocytes.
    J Virol. 1994 May;68(5):3410-4 PMID: 8151802
  67. Telomeres inhibit end to end fusion and enhance maintenance of linear DNA molecules injected into the Paramecium primaurelia macronucleus.
    Nucleic Acids Res. 1991 Apr 11;19(7):1541-7 PMID: 1851293
Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.134.1.1
Published
1996-07-00
Pages
1-12
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2120915
Subset
IM
Grants
NIA NIH HHS · AG-09383 · United States
External Links
PubMed source DOI: 10.1083/jcb.134.1.1 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2