A Quantitative Real-Time PCR Method for Absolute Telomere Length

O’Callaghan, Nathan; Dhillon, Varinderpal S.; Thomas, Philip; Fenech, Michael

doi:10.2144/000112761

Cited by 208 publications

(206 citation statements)

References 16 publications

(15 reference statements)

Supporting

Mentioning

203

Contrasting

Unclassified

Order By: Relevance

“…Telomere length assessment via qPCR has already been validated (Grabowski et al, 2005;O'Callaghan et al, 2008). However, although qPCR telomere measurements are normalized based on single copy genes in humans and other model species, the lack of genomic data for non-model species such as C. tenuispina hinders the identification of nuclear genes without paralogs for normalization.…”

Section: Telomere Measurementsmentioning

confidence: 99%

Long telomeres are associated with clonality in wild populations of the fissiparous starfish Coscinasterias tenuispina

et al. 2015

View full text Add to dashboard Cite

Telomeres usually shorten during an organism's lifespan and have thus been used as an aging and health marker. When telomeres become sufficiently short, senescence is induced. The most common method of restoring telomere length is via telomerase reverse transcriptase activity, highly expressed during embryogenesis. However, although asexual reproduction from adult tissues has an important role in the life cycles of certain species, its effect on the aging and fitness of wild populations, as well as its implications for the long-term survival of populations with limited genetic variation, is largely unknown. Here we compare relative telomere length of 58 individuals from four populations of the asexually reproducing starfish Coscinasterias tenuispina. Additionally, 12 individuals were used to compare telomere lengths in regenerating and non-regenerating arms, in two different tissues (tube feet and pyloric cecum). The level of clonality was assessed by genotyping the populations based on 12 specific microsatellite loci and relative telomere length was measured via quantitative PCR. The results revealed significantly longer telomeres in Mediterranean populations than Atlantic ones as demonstrated by the Kruskal-Wallis test (K = 24.17, significant value: P-valueo0.001), with the former also characterized by higher levels of clonality derived from asexual reproduction. Telomeres were furthermore significantly longer in regenerating arms than in non-regenerating arms within individuals (pyloric cecum tissue: Mann-Whitney test, V = 299, P-valueo10 − 6 ; and tube feet tissue Student's t = 2.28, P-value = 0.029). Our study suggests that one of the mechanisms responsible for the long-term somatic maintenance and persistence of clonal populations is telomere elongation.

show abstract

Section: Telomere Measurementsmentioning

confidence: 99%

Long telomeres are associated with clonality in wild populations of the fissiparous starfish Coscinasterias tenuispina

et al. 2015

View full text Add to dashboard Cite

show abstract

“…DNA was quantified in triplicate using a NanoDrop spectrophotometer (Thermo Fisher Scientific Inc. Waltham, MA USA). Each PCR (20 μL) was performed as follows: 20 ng DNA, 1 × SYBR ® Green master mix (Life Technologies‐Applied Biosystems Italia Monza, Italy), 100 n m telomere forward primer (CGG TTT GTT TGG GTT TGG GTT TGG GTT TGG GTT TGG GTT), and 100 n m telomere reverse primer (GGC TTG CCT TAC CCT TAC CCT TAC CCT TAC CCT TAC CCT) (O'Callaghan et al ., 2008). Results are from the measure of three individuals per group performed in quadruplicates.…”

Section: Methodsmentioning

confidence: 99%

P66SHC deletion improves fertility and progeric phenotype of late-generation TERC-deficient mice but not their short lifespan

2016

View full text Add to dashboard Cite

SummaryOxidative stress and telomere attrition are considered the driving factors of aging. As oxidative damage to telomeric DNA favors the erosion of chromosome ends and, in turn, telomere shortening increases the sensitivity to pro‐oxidants, these two factors may trigger a detrimental vicious cycle. To check whether limiting oxidative stress slows down telomere shortening and related progeria, we have investigated the effect of p66SHC deletion, which has been shown to reduce oxidative stress and mitochondrial apoptosis, on late‐generation TERC (telomerase RNA component)‐deficient mice having short telomeres and reduced lifespan. Double mutant (TERC −/− p66SHC −/−) mice were generated, and their telomere length, fertility, and lifespan investigated in different generations. Results revealed that p66SHC deletion partially rescues sterility and weight loss, as well as organ atrophy, of TERC‐deficient mice, but not their short lifespan and telomere erosion.Therefore, our data suggest that p66SHC‐mediated oxidative stress and telomere shortening synergize in some tissues (including testes) to accelerate aging; however, early mortality of late‐generation mice seems to be independent of any link between p66SHC‐mediated oxidative stress and telomere attrition.

show abstract

“…The telomere signal was normalized to the signal from the single copy gene to generate a T/S ratio indicative of relative telomere length. For absolute telomere length calculation [53], the telomere kb per reaction value was divided by diploid genome copy number (calculated from 36B4 Ct and standard curve) to generate a total telomeric length in kb per diploid genome. Equal amounts of DNA (20 ng) were used for each reaction, with at least three replicates for each specimen.…”

Section: Telomere Measurement By Quantitative Real-time Pcrmentioning

confidence: 99%

Association of telomere length with authentic pluripotency of ES/iPS cells

et al. 2011

View full text Add to dashboard Cite

Telomerase and telomeres are important for indefinite replication of stem cells. Recently, telomeres of somatic cells were found to be reprogrammed to elongate in induced pluripotent stem cells (iPSCs). The role of telomeres in developmental pluripotency in vivo of embryonic stem cells (ESCs) or iPSCs, however, has not been directly addressed. We show that ESCs with long telomeres exhibit authentic developmental pluripotency, as evidenced by generation of complete ESC pups as well as germline-competent chimeras, the most stringent tests available in rodents. ESCs with short telomeres show reduced teratoma formation and chimera production, and fail to generate complete ESC pups. Telomere lengths are highly correlated (r > 0.8) with the developmental pluripotency of ESCs. Short telomeres decrease the proliferative rate or capacity of ESCs, alter the expression of genes related to telomere epigenetics, down-regulate genes important for embryogenesis and disrupt germ cell differentiation. Moreover, iPSCs with longer telomeres generate chimeras with higher efficiency than those with short telomeres. Our data show that functional telomeres are essential for the developmental pluripotency of ESCs/iPSCs and suggest that telomere length may provide a valuable marker to evaluate stem cell pluripotency, particularly when the stringent tests are not feasible.

show abstract

A Quantitative Real-Time PCR Method for Absolute Telomere Length

Cited by 208 publications

References 16 publications

Long telomeres are associated with clonality in wild populations of the fissiparous starfish Coscinasterias tenuispina

Long telomeres are associated with clonality in wild populations of the fissiparous starfish Coscinasterias tenuispina

P66SHC deletion improves fertility and progeric phenotype of late-generation TERC-deficient mice but not their short lifespan

Association of telomere length with authentic pluripotency of ES/iPS cells

Contact Info

Product

Resources

About