Abstract
Structural and functional analysis of telomeres is very important for understanding basic biological functions such as genome stability, cell growth control, senescence and aging. Recently, serious concerns have been raised regarding the reliability of current telomere measurement methods such as Southern blot and quantitative polymerase chain reaction. Since telomere length is associated with age related pathologies, including cardiovascular disease and cancer, both at the individual and population level, accurate interpretation of measured results is a necessity. The telomere Q-PNA-FISH technique has been widely used in these studies as well as in commercial analysis for the general population. A hallmark of telomere Q-PNA-FISH is the wide variation among telomere signals which has a major impact on obtained results. In the present study we introduce a specific mathematical and statistical analysis of sister telomere signals during cell culture senescence which enabled us to identify high regularity in their variations. This phenomenon explains the reproducibility of results observed in numerous telomere studies when the Q-PNA-FISH technique is used. In addition, we discuss the molecular mechanisms which probably underlie the observed telomere behavior.
Highlights
Telomeres are specialized structures at the ends of linear chromosomes, composed of repetitive DNA and an associated protein complex called shelterin [1]
While telomere length followed through Southern blot and quantitative polymerase chain reaction (Q-PCR) analysis gives us plenty of information about their gross dynamics [10], it is very important to monitor the behavior of individual telomeres as well, especially when considering medical predictions or pharmaceutical effects [11,12]
Q-PNA-FISH using a C rich probe for labeling of the G rich telomere strand is the most common technique used to follow individual telomere dynamics in numerous publications. We employed this method to analyze sister telomere pairs on metaphase chromosomes of normal and hTERT MJ90 human fibroblasts with increasing population doublings (PDs)
Summary
Telomeres are specialized structures at the ends of linear chromosomes, composed of repetitive DNA and an associated protein complex called shelterin [1]. While telomere length followed through Southern blot and Q-PCR analysis gives us plenty of information about their gross dynamics [10], it is very important to monitor the behavior of individual telomeres as well, especially when considering medical predictions or pharmaceutical effects [11,12]. For these considerations, Q-PNA-FISH has become the method of choice
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