title: Rapid telomere attrition in cardiac tissue of the ageing Wistar rat.
abstract: Many studies show an association between ageing and mean telomere length in DNA isolated from peripheral blood mononuclear cells, few studies have examined less accessible tissues. This study has two objectives: (i) to define the best method to prepare rodent DNA for telomere length measurement by Southern blotting and (ii) to determine whether there are differential rates of telomere attrition in different rodent tissues. We found that the use of agarose plugs for DNA isolation was essential for the accurate measurement of rodent telomere length. Tissue was collected from neonatal (3 days) or aged (18-24 months) male Wistar rats and terminal restriction fragment (TRF) length was measured by Southern blotting. Cardiac tissue from aged rats showed a 38% loss of TRF length compared with newborn animals (p<0.001, n=13), this contrasts with much smaller reductions in brain (1.6%), liver (14.2%), kidney (8.9%) and lung (9.7%). This study demonstrates that the methods of DNA preparation are critical for accurate measurement of telomeres in rodent tissues. Moreover, we show differential rates of telomere attrition in rat tissues, the heart being most susceptible to telomere loss. These observations could have important implications for the study of age-specific changes in tissue function.
authors.affiliation: Department of Cardiovascular Sciences, University of Leicester, Robert Kilpatrick Clinical Sciences Building, Leicester Royal Infirmary, Leicester LE2 7LX, UK. rah22@le.ac.uk
authors.forename: Richard
authors.initials: R
authors.lastname: Hastings
authors.forename: Nai-Chang
authors.initials: NC
authors.lastname: Li
authors.forename: Peter S
authors.initials: PS
authors.lastname: Lacy
authors.forename: Hasmukh
authors.initials: H
authors.lastname: Patel
authors.forename: Karl E
authors.initials: KE
authors.lastname: Herbert
authors.forename: Adrian G
authors.initials: AG
authors.lastname: Stanley
authors.forename: Bryan
authors.initials: B
authors.lastname: Williams
mesh_terms: D000375:Aging / Q000502:physiology*; D000818:Animals; D015139:Blotting, Southern / Q000379:methods; D001921:Brain / Q000502:physiology; D004247:DNA / Q000032:analysis; D006321:Heart / Q000196:embryology / Q000502:physiology; D007668:Kidney / Q000502:physiology; D008099:Liver / Q000502:physiology; D008168:Lung / Q000502:physiology; D008297:Male; D009206:Myocardium; D051381:Rats; D017208:Rats, Wistar; D016615:Telomere / Q000235:genetics*
chemical_list: D004247:DNA