Abstract
The replicative lifespan (RLS) of a cell-defined as the number of cell divisions before death-has informed our understanding of the mechanisms of cellular aging. However, little is known about aging and longevity in symmetrically dividing eukaryotic cells because most prior studies have used budding yeast for RLS studies. Here, we describe a multiplexed fission yeast lifespan micro-dissector (multFYLM) and an associated image processing pipeline for performing high-throughput and automated single-cell micro-dissection. Using the multFYLM, we observe continuous replication of hundreds of individual fission yeast cells for over seventy-five generations. Surprisingly, cells die without the classic hallmarks of cellular aging, such as progressive changes in size, doubling time, or sibling health. Genetic perturbations and drugs can extend the RLS via an aging-independent mechanism. Using a quantitative model to analyze these results, we conclude that fission yeast does not age and that cellular aging and replicative lifespan can be uncoupled in a eukaryotic cell.
Highlights
Aging is the progressive decrease of an organism’s fitness over time
A high-throughput microfluidic assay for measuring the replicative lifespan in fission yeast
We recently addressed this challenge by developing a microfluidic platform for capturing and immobilizing individual fission yeast cells via their old cell tips [17]
Summary
Aging is the progressive decrease of an organism’s fitness over time. The asymmetric segregation of a set of molecules called pro-aging factors during mitosis has been proposed to promote aging in both yeast and in higher eukaryotes [1,2,3,4]. By sequestering pro-aging factors in the mothers, newly born daughters reset their RLS [3, 8, 10] These observations raise the possibility that alternate mechanisms may be active in symmetrically dividing eukaryotic cells. The fission yeast Schizosaccharomyces pombe is an excellent model system for investigating RLS and aging phenotypes in symmetrically dividing eukaryotic cells. Papers reported aging phenotypes akin to those observed in budding yeast (e.g., mother cells become larger, divide more slowly, and have less healthy offspring as they age) [2, 14]. We report the first high-throughput characterization of both RLS and aging in fission yeast To enable these studies, we developed the multiplexed fission yeast lifespan microdissector (FYLM) — a highthroughput microfluidic platform and software analysis suite that captures and tracks individual S. pombe cells throughout their RLS. We conclude that fission yeast dies primarily via a stochastic, age-independent mechanism
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