Monitoring Telomere Maintenance During Regeneration of Annelids

Methods Mol Biol. 2022:2450:467-478. doi: 10.1007/978-1-0716-2172-1_24.

Abstract

Telomere shortening is a hallmark of aging and eventually constrains the proliferative capacity of cells. The protocols discussed here are used for monitoring telomeres comprehensively in Aeolosoma viride, a model system for regeneration studies. We present methods for analyzing the activity of telomerase enzyme in regenerating tissue by telomeric repeat amplification protocol (TRAP) assay, for comparing telomere length between existing tissue and newly regenerated tissue by telomere restriction fragment (TRF) assay, as well as for visualizing telomeres by fluorescence in situ hybridization (FISH).

Keywords: Aeolosoma viride; Regeneration; TRAP assay; TRF assay; Telomere FISH; Telomere maintenance.

MeSH terms

  • Animals
  • Annelida*
  • In Situ Hybridization, Fluorescence
  • Telomerase* / genetics
  • Telomerase* / metabolism
  • Telomere / genetics
  • Telomere / metabolism
  • Telomere Homeostasis

Substances

  • Telomerase