Skip to main content
SpringerLink
Log in

Telomere Length Analysis

  • Protocol
Biological Aging

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 371))

Abstract

Most somatic cells of long-lived species undergo telomere shortening throughout life. Critically short telomeres trigger loss of cell viability in tissues, which has been related to alteration of tissue function and loss of regenerative capabilities in aging and aging-related diseases. Hence, telomere length is an important biomarker for aging and can be used in the prognosis of aging diseases. These facts highlight the importance of developing methods for telomere length determination that can be employed to evaluate telomere length during the human aging process. Telomere length quantification methods have improved greatly in accuracy and sensitivity since the development of the conventional telomeric Southern blot. Here, we describe the different methodologies recently developed for telomere length quantification, as well as their potential applications for human aging studies.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Blackburn, E. H. (2001) Switching and signaling at the telomere. Cell 106, 661–673.

    Article  CAS  PubMed  Google Scholar 

  2. de Lange, T. (2004) T-loops and the origin of telomeres. Nat. Rev. Mol. Cell Biol. 5, 323–329.

    Article  PubMed  Google Scholar 

  3. Smogorzewska, A. and de Lange, T. (2002) Different telomere damage signaling pathways in human and mouse cells. EMBO J. 21, 4338–4348.

    Article  CAS  PubMed  Google Scholar 

  4. d’ Adda di Fagagna, F., Reaper, P. M., Clay-Farrace, L., et al. (2003) A DNA damage checkpoint response in telomere-initiated senescence. Nature 426, 194–198.

    Article  Google Scholar 

  5. Takai, H., Smogorzewska, A., and de Lange, T. (2003) DNA damage foci at dysfunctional telomeres. Curr. Biol. 13, 1549–1556.

    Article  CAS  PubMed  Google Scholar 

  6. Olovnikov, A. M. (1973) A theory of marginotomy. The incomplete copying of template margin in enzymic synthesis of polynucleotides and biological significance of the phenomenon. J. Theor. Biol. 41, 181–190.

    Article  CAS  PubMed  Google Scholar 

  7. Shay, J. W. and Wright, W. E. (2005) Senescence and immortalization: role of telomeres and telomerase. Carcinogenesis 26, 867–874.

    Article  CAS  PubMed  Google Scholar 

  8. Collins, K. and Mitchell, J. R. (2002) Telomerase in the human organism. Oncogene 21, 564–579.

    Article  CAS  PubMed  Google Scholar 

  9. Allsopp, R. C., Vaziri, H., Patterson, C., et al. (1992) Telomere length predicts replicative capacity of human fibroblasts. Proc. Natl. Acad. Sci. USA 89, 10,114–10,118.

    Article  CAS  PubMed  Google Scholar 

  10. Kitada, T., Seki, S., Kawakita, N., Kuroki, T., and Monna, T. (1995) Telomere shortening in chronic liver diseases. Biochem. Biophys. Res. Commun. 211, 33–39.

    Article  CAS  PubMed  Google Scholar 

  11. Okuda, K., Khan, M. Y., Skurnick, J., Kimura, M., Aviv, H., and Aviv, A. (2000) Telomere attrition of the human abdominal aorta: relationships with age and atherosclerosis. Atherosclerosis 152, 391–398.

    Article  CAS  PubMed  Google Scholar 

  12. Jeanclos, E., Krolewski, A., Skurnick, J., et al. (1998) Shortened telomere length in white blood cells of patients with IDDM. Diabetes 47, 482–486.

    Article  CAS  PubMed  Google Scholar 

  13. Decary, S., Mouly, V., Hamida, C. B. Sautet, A., Barbet, J. P., and Butler-Browne, G. S. (1997) Replicative potential and telomere length in human skeletal muscle: implications for satellite cell-mediated gene therapy. Hum. Gene Ther. 8, 1429–1438.

    Article  CAS  PubMed  Google Scholar 

  14. Wright, W. E., Piatyszek, M. A., Rainey, W. E., Byrd, W., and Shay, J. W. (1996) Telomerase activity in human germline and embryonic tissues and cells. Dev. Genet. 18, 173–179.

    Article  CAS  PubMed  Google Scholar 

  15. von Zglinicki, T., Serra, V., Lorenz, M., et al. (2000) Short telomeres in patients with vascular dementia: an indicator of low antioxidative capacity and a possible risk factor? Lab. Invest. 80, 1739–1747.

    Article  Google Scholar 

  16. Samani, N. J., Boultby, R., Butler, R., Thompson, J. R., and Goodall, A. H. (2001) Telomere shortening in atherosclerosis. Lancet 358, 472–473.

    Article  CAS  PubMed  Google Scholar 

  17. Wu, X., Amos, C. I., Zhu, Y., et al. (2003) Telomere dysfunction: a potential cancer predisposition factor. J. Natl. Cancer Inst. 95, 1211–1218.

    Article  CAS  PubMed  Google Scholar 

  18. Blasco, M. A., Lee, H. W., Hande, M. P., et al. (1997) Telomere shortening and tumor formation by mouse cells lacking telomerase RNA. Cell 91, 25–34.

    Article  CAS  PubMed  Google Scholar 

  19. Lee, H. W., Blasco, M. A., Gottlieb, G. J., Horner, J. W., 2nd, Greider, C. W., and DePinho, R. A. (1998) Essential role of mouse telomerase in highly proliferative organs. Nature 392, 569–574.

    Article  CAS  PubMed  Google Scholar 

  20. Herrera, E., Samper, E., Martin-Caballero, J., Flores, J. M., Lee, H. W. and Blasco, M. A. (1999) Disease states associated with telomerase deficiency appear earlier in mice with short telomeres. EMBO J. 18, 2950–2960.

    Article  CAS  PubMed  Google Scholar 

  21. Herrera, E., Martinez, A. C., and Blasco, M. A. (2000) Impaired germinal center reaction in mice with short telomeres. EMBO J. 19, 472–481.

    Article  CAS  PubMed  Google Scholar 

  22. Herrera, E., Samper, E., and Blasco, M. A. (1999) Telomere shortening in mTR-/-embryos is associated with failure to close the neural tube. EMBO J. 18, 1172–1181.

    Article  CAS  PubMed  Google Scholar 

  23. Rudolph, K. L., Chang, S., Lee, H. W., et al. (1999) Longevity, stress response, and cancer in aging telomerase-deficient mice. Cell 96, 701–712.

    Article  CAS  PubMed  Google Scholar 

  24. Franco, S., Segura, I., Riese, H. H., and Blasco, M. A. (2002) Decreased B16F10 melanoma growth and impaired vascularization in telomerase-deficient mice with critically short telomeres. Cancer Res. 62, 552–559.

    CAS  PubMed  Google Scholar 

  25. Samper, E., Fernandez, P., Eguia, R., et al. (2002) Long-term repopulating ability of telomerase-deficient murine hematopoietic stem cells. Blood 99, 2767–2775.

    Article  CAS  PubMed  Google Scholar 

  26. Leri, A., Franco, S., Zacheo, A., et al. (2003) Ablation of telomerase and telomere loss leads to cardiac dilatation and heart failure associated with p53 upregulation. EMBO J. 22, 131–139.

    Article  CAS  PubMed  Google Scholar 

  27. Harley, C. B. (1991) Telomere loss: mitotic clock or genetic time bomb? Mutat. Res. 256, 271–282.

    CAS  PubMed  Google Scholar 

  28. Harley, C. B. (1995) Telomeres and aging, in Telomeres (Blackburn, E. H. and Greider, C. W., eds.). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY: pp. 247–264.

    Google Scholar 

  29. Nakamura, Y., Hirose, M., Matsuo, H., Tsuyama, N., Kamisango, K., and Ide, T. (1999) Simple, rapid, quantitative, and sensitive detection of telomere repeats in cell lysate by a hybridization protection assay. Clin. Chem. 45, 1718–1724.

    CAS  PubMed  Google Scholar 

  30. Freulet-Marriere, M. A., Potocki-Veronese, G., Deverre, J. R., and Sabatier, L. (2004) Rapid method for mean telomere length measurement directly from cell lysates. Biochem. Biophys. Res. Commun. 314, 950–956.

    Article  CAS  PubMed  Google Scholar 

  31. Therkelsen, A. J., Nielsen, A., Koch, J., Hindkjaer, J., and Kolvraa, S. (1995) Staining of human telomeres with primed in situ labeling (PRINS). Cytogenet. Cell Genet. 68, 115–118.

    Article  CAS  PubMed  Google Scholar 

  32. Baird, D. M., Rowson, J., Wynford-Thomas, D., and Kipling, D. (2003) Extensive allelic variation and ultrashort telomeres in senescent human cells. Nat. Genet. 33, 203–207.

    Article  CAS  PubMed  Google Scholar 

  33. Moyzis, R. K., Buckingham, J. M., Cram, L. S., et al. (1988) A highly conserved repetitive DNA sequence, (TTAGGG)n, present at the telomeres of human chromosomes. Proc. Natl. Acad. Sci. USA 85, 6622–6626.

    Article  CAS  PubMed  Google Scholar 

  34. Allshire, R. C. Gosden, J. R., Cross, S. H., et al. (1988) Telomeric repeat from T. thermophila cross hybridizes with human telomeres. Nature 332, 656–659.

    Article  CAS  PubMed  Google Scholar 

  35. Kipling, D. and Cooke, H. J. (1990) Hypervariable ultra-long telomeres in mice. Nature 347, 400–402.

    Article  CAS  PubMed  Google Scholar 

  36. Hemann, M. T. and Greider, C. W. (1999) G-strand overhangs on telomeres in telomerase-deficient mouse cells. Nucleic Acids Res. 27, 3964–3969.

    Article  CAS  PubMed  Google Scholar 

  37. Lansdorp, P. M., Verwoerd, N. P., van de Rijke, F. M., et al. (1996) Heterogeneity in telomere length of human chromosomes. Hum. Mol. Genet. 5, 685–691.

    Article  CAS  PubMed  Google Scholar 

  38. Nielsen, P. E., Egholm, M., Berg, R. H., and Buchardt, O. (1991) Sequence-selective recognition of DNA by strand displacement with a thymine-substituted polyamide. Science 254, 1497–1500.

    Article  CAS  PubMed  Google Scholar 

  39. Egholm, M., Buchardt, O., Christensen, L., et al. (1993) PNA hybridizes to complementary oligonucleotides obeying the Watson-Crick hydrogen-bonding rules. Nature 365, 566–568.

    Article  CAS  PubMed  Google Scholar 

  40. Poon, S. S., Martens, U. M., Ward, R. K., and Lansdorp, P. M. (1999) Telomere length measurements using digital fluorescence microscopy. Cytometry 36, 267–278.

    Article  CAS  PubMed  Google Scholar 

  41. Zijlmans, J. M., Martens, U. M., Poon, S. S., et al. (1997) Telomeres in the mouse have large inter-chromosomal variations in the number of T2AG3 repeats. Proc. Natl. Acad. Sci. USA 94, 7423–7428.

    Article  CAS  PubMed  Google Scholar 

  42. Martens, U. M., Zijlmans, J. M., Poon, S. S., et al. (1998) Short telomeres on human chromosome 17p. Nat. Genet. 18, 76–80.

    CAS  PubMed  Google Scholar 

  43. Samper, E., Flores, J. M., and Blasco, M. A. (2001) Restoration of telomerase activity rescues chromosomal instability and premature aging in Terc-/-mice with short telomeres. EMBO Rep. 2, 800–807.

    Article  CAS  PubMed  Google Scholar 

  44. Hemann, M. T., Strong, M. A., Hao, L. Y., and Greider, C. W. (2001) The shortest telomere, not average telomere length, is critical for cell viability and chromosome stability. Cell 107, 67–77.

    Article  CAS  PubMed  Google Scholar 

  45. Zou, Y., Sfeir, A., Gryaznov, S. M., Shay, J. W., and Wright, W. E. (2004) Does a sentinel or a subset of short telomeres determine replicative senescence? Mol. iol. Cell 15, 3709–3718.

    Article  CAS  Google Scholar 

  46. Hanish, J. P., Yanowitz, J. L., and de Lange, T. (1994) Stringent sequence requirements for the formation of human telomeres. Proc. Natl. Acad. Sci. USA 91, 8861–8865.

    Article  CAS  PubMed  Google Scholar 

  47. McIlrath, J., Bouffler, S. D., Samper, E., et al. (2001) Telomere length abnormalities in mammalian radiosensitive cells. Cancer Res. 61, 912–915.

    CAS  PubMed  Google Scholar 

  48. Gonzalez-Suarez, E., Samper, E., Flores, J. M., and Blasco, M. A. (2000) Telomerase-deficient mice with short telomeres are resistant to skin tumorigenesis. Nat. Genet. 26, 114–117.

    Article  CAS  PubMed  Google Scholar 

  49. Nagele, R. G., Velasco, A. Q., Anderson, W. J., et al. (2001) Telomere associations in interphase nuclei: possible role in maintenance of interphase chromosome topology. J. Cell Sci. 114, 377–388.

    CAS  PubMed  Google Scholar 

  50. Munoz, P., Blanco, R., Flores, J. M., and Blasco, M. A. (2005) XPF nuclease-dependent telomere loss and increased DNA damage in mice overexpressing TRF2 result in premature aging and cancer. Nat. Genet. 37, 1063–1071.

    Article  CAS  PubMed  Google Scholar 

  51. Rufer, N., Dragowska, W., Thornbury, G., Roosnek, E., and Lansdorp, P. M. (1998) Telomere length dynamics in human lymphocyte subpopulations measured by flow cytometry. Nat. Biotechnol. 16, 743–747.

    Article  CAS  PubMed  Google Scholar 

  52. Hultdin, M., Gronlund, E., Norrback, K., Eriksson-Lindstrom, E., Just, T., and Roos, G. (1998) Telomere analysis by fluorescence in situ hybridization and flow cytometry. Nucleic Acids Res. 26, 3651–3656.

    Article  CAS  PubMed  Google Scholar 

  53. Batliwalla, F. M., Damle, R. N., Metz, C. Chiorazzi, N., and Gregersen, P. K. (2001) Simultaneous flow cytometric analysis of cell surface markers and telomere length: analysis of human tonsilar B cells. J. Immunol. Methods 247, 103–109.

    Article  CAS  PubMed  Google Scholar 

  54. Baerlocher, G. M. and Lansdorp, P. M. (2003) Telomere length measurements in leukocyte subsets by automated multicolor flow-FISH. Cytometry A 55, 1–6.

    Article  PubMed  Google Scholar 

  55. Azzalin, C. M., Mucciolo, E., Bertoni, L., and Giulotto, E. (1997) Fluorescence in situ hybridization with a synthetic (T2AG3)n polynucleotide detects several intrachromosomal telomere-like repeats on human chromosomes. Cytogenet. Cell Genet. 78, 112–115.

    Article  CAS  PubMed  Google Scholar 

  56. Ouellette, M. M., Liao, M., Herbert, B. S., et al. (2000) Subsenescent telomere lengths in fibroblasts immortalized by limiting amounts of telomerase. J. Biol. Chem. 275, 10,072–10,076.

    Article  CAS  PubMed  Google Scholar 

  57. Franco, S., Alsheimer, M., Herrera, E., Benavente, R., and Blasco, M. A. (2002) Mammalian meiotic telomeres: composition and ultrastructure in telomerasedeficient mice. Eur. J. Cell Biol. 81, 335–340.

    Article  PubMed  Google Scholar 

  58. Garcia-Cao, M., O’Sullivan, R., Peters, A. H., Jenuwein, T., and Blasco, M. A. (2004) Epigenetic regulation of telomere length in mammalian cells by the Suv39h1 and Suv39h2 histone methyltransferases. Nat. Genet. 36, 94–99.

    Article  CAS  PubMed  Google Scholar 

  59. O’Sullivan, J. N., Bronner, M. P., Brentnall, T. A., et al. (2002) Chromosomal instability in ulcerative colitis is related to telomere shortening. Nat. Genet. 32, 280–284.

    Article  PubMed  Google Scholar 

  60. Hande, M. P., Samper, E., Lansdorp, P., and Blasco, M. A. (1999) Telomere length dynamics and chromosomal instability in cells derived from telomerase null mice. J. Cell Biol. 144, 589–601.

    Article  CAS  PubMed  Google Scholar 

  61. Espejel, S., Franco, S., Rodriguez-Perales, S., Bouffler, S. D., Cigudosa, J. C., and Blasco, M. A. (2002) Mammalian Ku86 mediates chromosomal fusions and apoptosis caused by critically short telomeres. EMBO J. 21, 2207–2219.

    Article  CAS  PubMed  Google Scholar 

  62. Baerlocher, G. M., Mak, J., Tien, T., and Lansdorp, P. M. (2002) Telomere length measurement by fluorescence in situ hybridization and flow cytometry: tips and pitfalls. Cytometry 47, 89–99.

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Telomeres and Telomerase Group, Molecular Oncology Program, Spanish National Cancer Center, Madrid, Spain

    Andrés Canela, Peter Klatt & María A. Blasco

Authors
  1. Andrés Canela
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Peter Klatt
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. María A. Blasco
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. Department of Biology, University of Alabama at Birmingham, Birmingham, AL

    Trygve O. Tollefsbol

Rights and permissions

Reprints and permissions

Copyright information

© 2007 Humana Press Inc., Totowa, NJ

About this protocol

Cite this protocol

Canela, A., Klatt, P., Blasco, M.A. (2007). Telomere Length Analysis. In: Tollefsbol, T.O. (eds) Biological Aging. Methods in Molecular Biology™, vol 371. Humana Press. https://doi.org/10.1007/978-1-59745-361-5_5

Download citation

  • DOI: https://doi.org/10.1007/978-1-59745-361-5_5

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-658-0

  • Online ISBN: 978-1-59745-361-5

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation