Lehmen, Tassia Fontana et al. (2013) International Worm Meeting "Effect of chronic treatment with amphetamine in an experimental model using Caenorhabditis elegans."

Fachinetto, Roselei, Lehmen, Tassia Fontana, Soares, Felix A A, Puntel, Bruna, Gubert, Priscila

[International Worm Meeting, 2013]

Caenorhabditis elegans, a free-living soil nematode, has already been explored as a valuable bioindicator, since it is one of the best-characterized animal models at the genetic, physiological, molecular, and developmental levels. It has also been proved that C. elegans serves as an excellent candidate for studying the development and functions of nervous system and neurotoxicology. The worms have well characterized dopaminergic system, as well as eight neurons of this system located along the body. Amphetamine (AMPH) is a stimulant drug of the central nervous system that acts broadly on various behaviors. Whereas multiple lines of evidence implicate dopamine receptors in the mechanism of action of AMPH and other abused psychostimulants, the degree to which different receptors support AMPH-induced behaviors has not been completely elucidated. Because of this we decided to investigate the effects of amphetamine on the behavior of nematodes.L1 larva stage worms of wild type strain were treated with AMPH 1mM or water (control group) in agar plates until adulthood. We analyzed pharyngeal pumping rate, defecation cycle length, body bends and oviposition and egg production rates in adult worms using a microscope. Treatment with AMPH 1 mM promoted a reduction in the rates of pharyngeal pumping, oviposition and egg production. Our data demonstrates that the concentration of amphetamine tested can interfere the behaviors mentioned above without, however, changing the defecation cycle length and body bends, acting probably by different routes. However, more studies are needed to elucidate the different mechanisms of action that may be involved, using, for example, knockout strains to dopamine transporters and/or receptors.

Coleman-Hulbert, A.L. et al. (2017) International Worm Meeting "Automation of the Caenorhabditis Intervention Testing Program."

Garret, T., Fulger, A.C., Lithgow, G.J., Johnson, E., Kish, J.L., Banse, S., Sedore, C.A., Presley, M.P., Driscoll, M., Harinath, G., Phillips, P.C., Lucanic, M., Falkowski, R., Chen, E., Blue, B.W., Crist, A.B., Hall, D., Coleman-Hulbert, A.L., Plummer, T.W.

[International Worm Meeting, 2017]

The Caenorhabditis Intervention Testing Program (CITP) is a multi-institutional consortium supported by the National Institutes of Aging and tasked with identifying robust life-extending compounds using nematodes from the genus Caenorhabditis. CITP partners at Rutgers University, the University of Oregon, and the Buck Institute for Aging Research have streamlined efforts and reproduced findings from 22 diverse strains of Caenorhabditis under ten separate compound interventions. Our work highlights the necessity of replication of longevity analysis, and our intention is to greatly expand the scale of our assays in order to keep up with demand from the aging research community. The C. elegans Lifespan Machine developed by Nicholas Stroustrup (Fontana Lab, Harvard) holds a great deal of promise as a high-throughput longevity approach. The C. elegans Lifespan Machine uses typical office scanners modified to hold petri plates containing nematodes. Through hourly image capture and later image processing, the Lifespan Machine automates the process of gathering lifespan data while providing tight temporal scaling. Here, we compare manual and automated lifespan measurements for multiple compounds across dozens of strains within each of the three laboratories. Overall, there are distinct differences in phenotype between manual and automated approaches for some strains and, especially, for some compounds. Lab to lab variation can also be a challenge. Nevertheless, the automated approach provides a great deal of value to the workflow of the CITP, and we plan to incorporate it into our future work, coupled with verification via manual lifespan assays.

Schiffer, Jodie et al. (2017) International Worm Meeting "How do worms determine their readiness to cope with environmental stress?"

Stumbur, Stephanie, Heath, William, Tam, Hannah, McGowan, Natalie, Vogelaar, Abigail, Schiffer, Jodie, Apfeld, Javier, Stanley, Julian

[International Worm Meeting, 2017]

At any time in their life, worms may encounter harmful environmental conditions such as high concentrations of oxidants or high temperature. We are interested in understanding how worms prepare for these potentially lethal events. We want to know: what establishes how prepared they are? To determine the mechanisms that control survival under harmful conditions, we are examining the effects of strong loss-of-function and null mutants in a collection of intercellular signaling receptors and transcription factors regulated by these receptors. We measure survival under oxidative stress and at high temperature using a "Lifespan Machine" cluster of flat-bed scanners [1]. This automated technology presents substantial advancements in throughput and sensitivity compared to manual methods. Using this approach, we have identified several signaling receptors and transcription factors that regulate survival under oxidative conditions. Interestingly, we identified both receptors and transcription factors that function to either confer or limit oxidative-stress resistance. This indicates that the combined level of activity of these genes sets the worm's readiness to cope with oxidative stress. We are currently investigating whether these genes act together or independently to determine the worm's normal level of resistance to various stressors, and the mechanisms that establish the normal activity levels of each of these genetic determinants of stress resistance. Reference: 1. Stroustrup N, Ulmschneider BE, Nash ZM, Lopez-Moyado IF, Apfeld J, Fontana W. The Caenorhabditis elegans Lifespan Machine. Nature Methods. 2013;10(7):665-70.

Garrett, Theo et al. (2017) International Worm Meeting "Ceanorhabditis Intervention Testing Program: Using Automated Healthspan Assays to screen for Chemicals that Mitigate Age-Dependent Declines in Stress Resistance."

Presley, Michael, Driscoll, Monica, Garrett, Theo, Lucanic, Mark, Lithgow, Gordon, Phillips, Patrick, Hall, David, Plummer, W. Todd, Foulger, Anna

[International Worm Meeting, 2017]

The Caenorhabditis Intervention Testing Program (CITP) is a National Institutes on Aging supported consortium that screens promising chemicals across diverse genetic backgrounds for effects on lifespan and healthspan. Such chemicals are desirable, as they are likely to target conserved pathways that modulate aging. Pharmacological interventions that target cor aging pathways are expected to be useful in treating multiple human aging related diseases. We utilize the model Caenorhabditis elegans, as its relatively short lifespand and predictable responses to stress allow for high-throughput screening with large sample sizes. To increase the data collection capacity of our measurements we have utilized the Automated Lifespan Machines, developed by the Fontana Lab 1. Here we describe our results using these machines to study not lifespan but healthspan. Specifically, we will present our results from testing chemical treatments for their ability to mitigate age-dependent declines in stress resistance. We first characterized the age dependent declines of multiple strains and species in both thermal and oxidative stress resistance. We then go on to describe the results of two reported pro-longevity chemicals on mitigating these declines. We describe our results for Rapamycin, an mTOR inhibitor that has shown robust pro-longevity effects across multiple studies, and Acarbose, an alpha-glucosidase inhibitor. The CITP's novel combination of testing diverse genetic backgrounds, use of computer controlled scanners, and devotion to mulit-replicate assays has allowed the CITP to confidently assess the effects of pro-longevity chemicals for their ability to mitigate age-dependent declines in stress resistance. We suggest that this activity may be at least as pharmacologically important as the chemicals' ability to extend lifespan.