Invertebrate Longevity and Healthspan

The Invertebrate Longevity and Healthspan Core assists investigators throughout the research community in quantifying a variety of longevity and healthspan measures in two of the major invertebrate model organisms: the budding yeast Saccharomyces cerevisiae and the nematode Caenorhabditis elegans. This is accomplished through direct collaboration, training and outreach, and the development and dissemination of new tools and technologies. New technologies we develope and disseminate include microfluidic tools for invertebrate aging studies and the WormBot, a robotic system for high-throughput survival and behavioral phenotyping in C. elegans.  We also work to develop chemostat-based solutions for obtaining age-matched pure populations of yeast for biochemical and cell biological experiments requested by Core users.  The focus on yeast and worms allows us to provide services and technologies to a large number of external investigators and thereby have a correspondingly large impact.  The data generated by this Core can often be correlated with high dimensional computational analyses and with the protein and metabolite phenotypes studied in the other Nathan Shock Center Cores. Invertebrate models are also highly accessible to junior investigators, and our Center pilot projects allow these investigators access to the full range of integrated services provided by the Center Resource Cores.

Services Available:

The following services are available to all users.  Additional services may also be available and users are encouraged to contact the Core Director to discuss specific experimental details.  Typical costs are provided below for initial budget preparation and may be used for Pilot Project submissions, but in all cases final costs will be determined following consultation with Core personnel.

Saccharomyces cerevisiae

Yeast aging chemostat experiments. Chemostats can be used to generate large number of aged cells for a number of downstream applications. Cost: dependent on experiment. Training in chemostat construction and operation is also possible.

Caenorhabditis elegans 

1. Life span analysis.  The Core will perform life span studies on C. elegans strains using our standard NGM plate method [7]. Cost: $1500/12-well plate for industrial/ $750 for academic/internal.
2. Healthspan measures.  The Core will assay a variety of age-associated healthspan measures, including:  lipofuscin accumulation, muscle function, motility, resistance to polyglutamine or amyloid beta toxicity, and reproductive capacity (brood size).  Additional assays may be available upon request. Cost:  Cost will vary on a case-by-case basis, depending on services requested.
3. WormBot assays.  The Core will perform lifespan and healthspan assessments using the WormBot platform [8].  Cost: Estimated costs are $500 per plate.  Each plate can accommodate 12 experiments of 20-40 animals per experiment. 
4. Resistance/sensitivity to environmental or chemical stresses.  The core will perform survival assays in the presence of different environmental or chemical stressors upon request.  Cost:  Cost will vary on a case-by-case basis, depending on services requested.

General and Other Services

1. Access to organism-specific collections and libraries. The core will provide users with strains from the yeast ORF deletion collections, overexpression libraries, and GFP libraries for aging-related studies.  RNAi clones from the C. elegans RNAi libraries are also available.  Cost:  Access to libraries is offered free of charge when additional core services are requested.   
2. Construction and verification of deletion and/or transgenic strains.  Core personnel will consult with users on the best strategies for constructing strains of interest that are not currently available.  A variety of tools are available for these purposes, including plasmid collections for gene overexpression and DNA templates for PCR gene disruption using auxotrophic markers, and CRISPR and MosSCI systems for nematodes. The Core will make these tools available to users.  If desired, core personnel will also construct the strains for users. Cost:  $3000 per custom CRISPR edit or $1500 per overexpression extrachromosomal array.
3. Microscopy for C. elegans and S. cerevisiae. $1000 per microscope session per experimental group, including one experimental control group. 5-10 individual C. elegans in one field of view each per group. 5 fields of view per yeast group (each FOV can include dozens to hundreds of yeast cells depending on magnification and cell density).
4. WormBot System.  No custom microfluidics. Microfluidic C. elegans mounting devices available upon request

Literature Cited:

1.         Steffen, K.K., B.K. Kennedy, and M. Kaeberlein, Measuring replicative life span in the budding yeast. J Vis Exp, 2009(28).

2.         Lee, M.B., et al., A system to identify inhibitors of mTOR signaling using high-resolution growth analysis in Saccharomyces cerevisiae. Geroscience, 2017. 39(4): p. 419-428.

3.         Delaney, J.R., et al., Stress profiling of longevity mutants identifies Afg3 as a mitochondrial determinant of cytoplasmic mRNA translation and aging. Aging Cell, 2013. 12(1): p. 156-66.

4.         Olsen, B., C.J. Murakami, and M. Kaeberlein, YODA: software to facilitate high-throughput analysis of chronological life span, growth rate, and survival in budding yeast. BMC Bioinformatics, 2010. 11: p. 141.

5.         Crane, M.M., et al., DNA damage checkpoint activation impairs chromatin homeostasis and promotes mitotic catastrophe during aging. Elife, 2019. 8.

6.         Chen, K.L., et al., Loss of vacuolar acidity results in iron-sulfur cluster defects and divergent homeostatic responses during aging in Saccharomyces cerevisiae. Geroscience, 2020.

7.         Sutphin, G.L. and M. Kaeberlein, Measuring Caenorhabditis elegans life span on solid media. J Vis Exp, 2009(27).

8.         Pitt, J.N., et al., WormBot, an open-source robotics platform for survival and behavior analysis in C. elegans. Geroscience, 2019. 41(6): p. 961-973.

Core Personnel: The Core is led by Maitreya Dunham and Alex Mendenhall