Real-time quantitative PCR assay for measurement of avian telomeres

Francois Criscuolo*, Pierre Bize, Lubna Nasir, Neil B. Metcalfe, Chris G. Foote, Kate Griffiths, Elizabeth A. Gault, Pat Monaghan

*Corresponding author for this work

Research output: Contribution to journalArticle

126 Citations (Scopus)

Abstract

We present the application of a real-time quantitative PCR assay, previously developed to measure relative telomere length in humans and mice, to two bird species, the zebra finch Taeniopygia guttata and the Alpine swift Apus melba. This technique is based on the PCR amplification of telomeric (TTAGGG)(n) sequences using specific oligonucleotide primers. Relative telomere length is expressed as the ratio (T/S) of telomere repeat copy number (T) to control single gene copy number (S). This method is particularly useful for comparisons of individuals within species, or where the same individuals are followed longitudinally. We used glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as a single control gene. In both species, we validated our PCR measurements of relative telomere length against absolute measurements of telomere length determined by the conventional method of quantifying telomere terminal restriction fragment (TRF) lengths using both the traditional Southern blot analysis (Alpine swifts) and in gel hybridization (zebra finches). As found in humans and mice, telomere lengths in the same sample measured by TRF and PCR were well correlated in both the Alpine swift and the zebra finch.. Hence, this PCR assay for measurement of bird telomeres, which is fast and requires only small amounts of genomic DNA, should open new avenues in the study of environmental factors influencing variation in telomere length, and how this variation translates into variation in cellular and whole organism senescence.

Original languageEnglish
Pages (from-to)342-347
Number of pages6
JournalJournal of Avian Biology
Volume40
Issue number3
Early online date13 May 2009
DOIs
Publication statusPublished - May 2009

Keywords

  • LONG-LIVED BIRDS
  • LENGTH
  • CELLS
  • AGE
  • STRESS
  • DNA

Cite this

Criscuolo, F., Bize, P., Nasir, L., Metcalfe, N. B., Foote, C. G., Griffiths, K., Gault, E. A., & Monaghan, P. (2009). Real-time quantitative PCR assay for measurement of avian telomeres. Journal of Avian Biology, 40(3), 342-347. https://doi.org/10.1111/j.1600-048X.2008.04623.x