Telomerase activity is restricted in humans. Consequentially, telomeres shorten in most cells throughout our lives. Telomere dysfunction in vertebrates has been primarily studied in inbred mice strains with very long telomeres that fail to deplete telomeric repeats during their lifetime. It is, therefore, unclear how telomere shortening regulates tissue homeostasis in vertebrates with naturally short telomeres. Zebrafish have restricted telomerase expression and human-like telomere length. Here we show that first-generation tert^−/− zebrafish die prematurely with shorter telomeres. tert^−/− fish develop degenerative phenotypes, including premature infertility, gastrointestinal atrophy, and sarcopaenia. tert^−/− mutants have impaired cell proliferation, accumulation of DNA damage markers, and a p53 response leading to early apoptosis, followed by accumulation of senescent cells. Apoptosis is primarily observed in the proliferative niche and germ cells. Cell proliferation, but not apoptosis, is rescued in tp53^−/−tert^−/− mutants, underscoring p53 as mediator of telomerase deficiency and consequent telomere instability. Thus, telomerase is limiting for zebrafish lifespan, enabling the study of telomere shortening in naturally ageing individuals. Telomerase mutations in humans give rise to premature ageing syndromes. In animals, the wealth of knowledge in telomere biology has been biased by the almost exclusive analysis of long-telomere mice. The role of telomere shortening requires investigation in organisms that, much like humans, have evolved telomere length as an internal cell division “timer.” We provide evidence for such a model. We show for the first time that telomerase is required during zebrafish lifespan. In contrast to mice, first-generation telomerase zebrafish mutants display degenerative phenotypes and die prematurely by one year of age. Furthermore, we show that most telomerase deficiency in this model leads to time- and tissue-specific apoptotic and senescence responses, highlighting different tissue thresholds to telomere dysfunction. Our results show that telomeres are maintained just above a critical threshold and that telomerase function is truly limiting for zebrafish lifespan and tissue homeostasis, closely mimicking the human scenario.

Interleukin-7 (IL-7) is an essential cytokine for T-cell development and homeostasis. It is well established that IL-7 promotes the transcriptional down-regulation of IL7RA, leading to decreased IL-7Rα surface expression. However, it is currently unknown whether IL-7 regulates the intracellular trafficking and early turnover of its receptor on ligand binding. Here, we show that, in steady-state T cells, IL-7Rα is slowly internalized and degraded while a significant fraction recycles back to the surface. On IL-7 stimulation, there is rapid IL-7Rα endocytosis via clathrin-coated pits, decreased receptor recycling, and accelerated lysosome and proteasome-dependent degradation. In accordance, the half-life of IL-7Rα decreases from 24 hours to approximately 3 hours after IL-7 treatment. Interestingly, we further demonstrate that clathrin-dependent endocytosis is necessary for efficient IL-7 signal transduction. In turn, pretreatment of T cells with JAK3 or pan-JAK inhibitors suggests that IL-7Rα degradation depends on the activation of the IL-7 signaling effector JAK3. Overall, our findings indicate that IL-7 triggers rapid IL-7Rα endocytosis, which is required for IL-7–mediated signaling and subsequent receptor degradation.

The Smurf Assay (SA) was initially developed in the model organism Drosophila melanogaster where a dramatic increase of intestinal permeability has been shown to occur during aging (Rera et al., 2011). We have since validated the protocol in multiple other model organisms (Dambroise et al., 2016) and have utilized the assay to further our understanding of aging (Tricoire and Rera, 2015; Rera et al., 2018). The SA has now also been used by other labs to assess intestinal barrier permeability (Clark et al., 2015; Katzenberger et al., 2015; Barekat et al., 2016; Chakrabarti et al., 2016; Gelino et al., 2016). The SA in itself is simple; however, numerous small details can have a considerable impact on its experimental validity and subsequent interpretation. Here, we provide a detailed update on the SA technique and explain how to catch a Smurf while avoiding the most common experimental fallacies. Bio-protocol is an online peer-reviewed protocol journal. Its mission is to make life science research more efficient and reproducible by curating and hosting high quality, free access protocols.

Telomeres shorten with each cell division and telomere dysfunction is a recognized hallmark of aging. Tissue proliferation is expected to dictate the rate at which telomeres shorten. We set out to test whether proliferative tissues age faster than non-proliferative due to telomere shortening during zebrafish aging. We performed a prospective study linking telomere length to tissue pathology and disease. Contrary to expectations, we show that telomeres shorten to critical lengths only in specific tissues and independently of their proliferation rate. Short telomeres accumulate in the gut but not in other highly proliferative tissues such as the blood and gonads. Notably, the muscle, a low proliferative tissue, accumulates short telomeres and DNA damage at the same rate as the gut. Together, our work shows that telomere shortening and DNA damage in key tissues triggers not only local dysfunction but also anticipates the onset of age-associated diseases in other tissues, including cancer. Why, and how, organisms age and ultimately die is a key question of modern biology. Telomeres are considered molecular timekeepers determining cellular lifespans. Within an organism, tissue proliferation is expected to dictate the rate at which telomeres shorten. Using zebrafish, an organism with human-like telomeres, we set out to test whether, in natural aging, proliferative tissues age faster than non-proliferative in a telomere-dependent manner. We found that telomeres shorten with age to a level where they trigger telomere associated DNA damage and culminate in tissue dysfunction, independently of high or low tissue proliferation rates. Specifically, short telomeres accumulate in the gut, a highly proliferative tissue, and in the muscle, a low proliferative tissue, working as direct predictors of cellular damage prior to onset of intestinal inflammation and myocyte degeneration. Based on our data, we propose a model where telomere shortening in these key tissues is sufficient to trigger damage in others and precedes the onset of organism age-associated diseases, namely cancer. Thus, tissue-specific telomere length is limiting for local and systemic physiological integrity, leading to tissue degeneration and disease in aging.

Over 60% of people aged over 65 are affected by multiple morbidities, which are more difficult to treat, generate increased healthcare costs and lead to poor quality of life compared to individual diseases. With the number of older people steadily increasing this presents a societal challenge. Age is the major risk factor for age-related diseases and recent research developments have led to the proposal that pharmacological interventions targeting common mechanisms of ageing may be able to delay the onset of multimorbidity. Here we review the state of the knowledge of multimorbidity, appraise the available evidence supporting the role of mechanisms of ageing in the development of the most common age-related diseases and assess potential molecules that may successfully target those key mechanisms.