Microfluidic technologies for yeast replicative lifespan studies

Chen, Kenneth L.; Crane, Matthew M.; Kaeberlein, Matt

doi:10.1016/j.mad.2016.03.009

Cited by 67 publications

(48 citation statements)

References 59 publications

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“…Cells are typically maintained at 30°C during the day and micro-dissected every 2-3 hours, then placed at 4°C overnight to prevent overgrowth. More recently, several microfluidic methods for assessing RLS have been described in which mother cells are physically constrained in a microfluidic device such that fluid flow washes the daughter cells away from the mother cells (Chen et al, 2016). Both YPD and synthetic complete (SC) media types are routinely used for microfluidic RLS experiments.…”

Section: Dietary Restriction In S Cerevisiaementioning

confidence: 99%

Dietary restriction and lifespan: Lessons from invertebrate models

Kapahi

Kaeberlein

Hansen

2017

Ageing Research Reviews

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299

217

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Dietary restriction (DR) is the most robust environmental manipulation known to increase active and healthy lifespan in many species. Despite differences in the protocols and the way DR is carried out in different organisms, conserved relationships are emerging among multiple species. Elegant studies from numerous model organisms are further defining the importance of various nutrient-signaling pathways including mTOR (mechanistic target of rapamycin), insulin/IGF-1-like signaling and sirtuins in mediating the effects of DR. We here review current advances in our understanding of the molecular mechanisms altered by DR to promote lifespan in three major invertebrate models, the budding yeast Saccharomyces cerevisiae, the nematode Caenorhabditis elegans, and the fruit fly Drosophila melanogaster.

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Section: Dietary Restriction In S Cerevisiaementioning

confidence: 99%

Dietary restriction and lifespan: Lessons from invertebrate models

Kapahi

Kaeberlein

Hansen

2017

Ageing Research Reviews

Self Cite

299

217

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“…mother cells (22,23). Common pathways influence aging in yeast and multicellular eukaryotes (24,25), including caloric restriction (26), the mechanistic target of rapamycin (27), sirtuins (28), proteasomal function (29), and mRNA translation (30).…”

Section: Significancementioning

confidence: 99%

“…Colonies from independent spores served as replica cultures for fluctuation analysis using Can R [WT, n = 32 (combined from both genetic back-grounds); pol2-4, n = 14; msh6Δ, (combined from both genetic backgrounds) n = 15; pol3-01, n = 22; pol2-4 msh6Δ, n = 19]. The pol3-01 msh6Δ spores failed to form colonies; mutation rate was estimated from the synthetic relationship between proofreading and MMR in haploids(23) and confirmed by measurements of pol3-01/pol3-01 msh6Δ/msh6Δ diploids (SI Appendix,Fig. S1).…”

mentioning

confidence: 99%

Defining the impact of mutation accumulation on replicative lifespan in yeast using cancer-associated mutator phenotypes

Lee

Dowsett

Carr

et al. 2019

Proc. Natl. Acad. Sci. U.S.A.

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Mutations accumulate within somatic cells and have been proposed to contribute to aging. It is unclear what level of mutation burden may be required to consistently reduce cellular lifespan. Human cancers driven by a mutator phenotype represent an intriguing model to test this hypothesis, since they carry the highest mutation burdens of any human cell. However, it remains technically challenging to measure the replicative lifespan of individual mammalian cells. Here, we modeled the consequences of cancer-related mutator phenotypes on lifespan using yeast defective for mismatch repair (MMR) and/or leading strand (Polε) or lagging strand (Polδ) DNA polymerase proofreading. Only haploid mutator cells with significant lifetime mutation accumulation (MA) exhibited shorter lifespans. Diploid strains, derived by mating haploids of various genotypes, carried variable numbers of fixed mutations and a range of mutator phenotypes. Some diploid strains with fewer than two mutations per megabase displayed a 25% decrease in lifespan, suggesting that moderate numbers of random heterozygous mutations can increase mortality rate. As mutation rates and burdens climbed, lifespan steadily eroded. Strong diploid mutator phenotypes produced a form of genetic anticipation with regard to aging, where the longer a lineage persisted, the shorter lived cells became. Using MA lines, we established a relationship between mutation burden and lifespan, as well as population doubling time. Our observations define a threshold of random mutation burden that consistently decreases cellular longevity in diploid yeast cells. Many human cancers carry comparable mutation burdens, suggesting that while cancers appear immortal, individual cancer cells may suffer diminished lifespan due to accrued mutation burden.

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“…Because of this laborious process, the sample sizes are often too small to generate statistically meaningful results. Microfluidics in combination with automated microscopy, in contrast, allows lineages to be traced and the lifespan of large number of individual yeast strains to be accurately measured [17][19]. These measurements could potentially facilitate understandings of functions of genes and networks in development and aging.…”

Section: Facilitating Measurements and Enhancing Precision Of Perturbmentioning

confidence: 99%

Microfluidics in systems biology — hype or truly useful?

Liu

2016

Current Opinion in Biotechnology

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Systems biology often relies on large-scale measurements and to build models to understand how complex biological systems function. Microfluidic technology has been touted as a tool for high-throughput experiments and has been a valuable tool to some systems biology research. This review focuses on applications where microfluidics can enhance experimental sensitivity and throughput, particularly in recent development in single-cell analyses and analyses on multi-cellular or complex biological entities. We conclude that microfluidics is not necessarily always useful for systems biology, but when used appropriately can greatly enhance experimentalists’ ability to measure and control, and thereby enhance the understanding of and expand the utility of biological systems.

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Microfluidic technologies for yeast replicative lifespan studies

Cited by 67 publications

References 59 publications

Dietary restriction and lifespan: Lessons from invertebrate models

Dietary restriction and lifespan: Lessons from invertebrate models

Defining the impact of mutation accumulation on replicative lifespan in yeast using cancer-associated mutator phenotypes

Microfluidics in systems biology — hype or truly useful?

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