Longitudinal imaging of Caenorhabditis elegans in a microfabricated device reveals variation in behavioral decline during aging
- School of Engineering and Applied Sciences, University of Pennsylvania, United States
- Perelman School of Medicine, University of Pennsylvania, United States
Figures
Figure 1 with 4 supplements
WorMotel design, fabrication, preparation, and experimental setup.
(a) 3D rendering of WorMotel geometry. (b) Schematic cross-section of a single well. (c) Fabrication and loading process (d) Image of WorMotel filled with agar, bacteria, and adult C. elegans. (e) …
Figure 1—figure supplement 1
Comparison of image quality with standard 384-well plate and WorMotel.
(a) Image of L4 worms on agar in a standard 384-well plate. Scale bar: 2 mm. (b) Image of L4 worms on agar in a WorMotel. Individual animals are clearly visible in (b) but not in (a).
Figure 1—figure supplement 2
240-Well WorMotel (a) 240-well WorMotel PDMS insert.
Scale Bar: 1 cm. (b) PDMS insert placed inside an OmniTray. (c) Computer rendering of the 240-well WorMotel negative master (d) Image of a WorMotel filled with agar, bacteria, and worms in an …
Figure 1—figure supplement 3
WorMotel prevents burrowing.
Percent of animals burrowing in the WorMotel (red squares, n = 96) and in a standard 384-well microplate (blue circles, n = 80).
Figure 1—figure supplement 4
Schematic of automated imaging system.
An automated plate carousel (Thermo) containing eight plate stacks holds up to 240 WorMotel plates, each containing 240 C. elegans. A plate handling robot (Thermo Orbitor, ThermoFisher Scientific, …
Figure 2
Image processing, automated lifespan calculation, and manual validation.
(a) Activity calculation. Delta image (right image) is calculated by pixel-by-pixel subtraction of images taken at time t1 from images taken at time t2. Examples are shown in which a worm moves …
Figure 3 with 1 supplement
Automated quantification of behavioral changes during aging.
(a) Spontaneous behavior heat map for N2 (n = 445). Color bar indicates the number of pixels changed over a 60-s interval. (b) Stimulated behavior heat map for N2. (c) Spontaneous behavior heat map …
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Figure 3—source data 1
Includes data for lifespan, spontaneous locomotion, stimulated locomotion, response duration, and response latency for each strain shown.
- https://doi.org/10.7554/eLife.26652.014
Figure 3—figure supplement 1
Additional behavior heat maps.
(a) Spontaneous behavior heat map for age-1 mutants. (b) Stimulated behavior heat map for age-1 mutants. (c) Survivor spontaneous (dashed curve) and stimulated (solid curve) activity for age-1 …
Figure 4
Potential aging models.
(a) Model 1: Temporal scaling results in identical patterns of behavioral decline when data are normalized by lifespan. Idealized decline curves for wild type (black dot-dashed), short-lived (red …
Figure 5 with 1 supplement
Mutants with short and long lifespan display patterns of late-life behavioral decline that resemble short and long-lived worms from a wild type population.
(a) N2 behavior over time for the lowest quartile (solid curve) and highest quartile (dashed curve) of survivors. (b) Data from panel (a) plotted as a fraction of each individual’s life. (c) daf-16 …
Figure 5—figure supplement 1
Stimulated activity versus chronological time and fraction of life for long-lived individuals and short-Lived individuals of the same strain.
(a) age-1 mutant behavior as a function of chronological time for the lowest (solid curve) and highest (dashed curve) survivor quartiles. (b) tax-4 mutant behavior as a function of chronological …
Figure 6 with 1 supplement
Shape of behavioral decline changes continuously with lifespan across individuals and strains.
(a) Change in decline rate (pixels/life fraction) versus lifespan for individual N2 animals. (b) Same data as in panel (a) presented for daf-16 mutants. (c) Same data as in panel (a) presented for da…
Figure 6—figure supplement 1
Change in decline rate versus lifespan in individual animals.
(a) Change in decline rate versus lifespan for individuals of all strains tested. (b) Change in decline rate versus lifespan for individual age-1 mutants. (c) Same data as in panel (b) presented for …
Figure 7
Behavioral decline during acute oxidative stress resembles behavioral decline during aging.
(a, c, e, g) Behavior and survival heat maps for N2 (n = 58), daf-16 (n = 60), daf-2 (n = 68), and age-1 (n = 68). (b, d, f, h) Comparison of behavioral decline during normal aging (solid curve), …
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Figure 7—source data 1
Includes activity and paraquat survival data for each strain shown.
- https://doi.org/10.7554/eLife.26652.023
Figure 8
Reduced sensory response is not required for extended longevity in daf-2 mutants.
(a) Spontaneous activity during days 1–10 and 10–20 of adulthood for N2 grown on Empty Vector RNAi (n = 80), N2 grown on odr-10 RNAi (n = 40), daf-2 grown on Empty Vector RNAi (n = 40), and daf-2 …
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Figure 8—source data 1
Includes data for lifespan, spontaneous locomotion, stimulated locomotion, response duration, and response latency for each strain shown.
- https://doi.org/10.7554/eLife.26652.026
Videos
Video 1
Young (day 1) adults in the 240-well WorMotel.
A 10 s long blue light exposure (bright flash) occurs at the 15 min mark. FOV: 95 mm x 63 mm
Video 2
Detail of 9 wells containing day 1 N2 adults.
A 10 s long blue light exposure (bright flash) occurs at the 15 min mark. FOV: 14 mm x 14 mm
Video 3
Detail of 9 wells containing day 10 N2 adults.
A 10 s long blue light exposure (bright flash) occurs at the 15 min mark. FOV: 14 mm x 14 mm
Video 4
High resolution video of 9 WorMotel wells containing adult animals (day 1–4).
FOV: 14 mm x 14 mm
Tables
Table 1
Summary of lifespan data
| Strain | Food source | Lifespan (Mean ± SD) (Days) | N | P-value relative to N2 control |
|---|---|---|---|---|
| N2 | DA837 | 16.8 ± 4.4 | 445 | N/A |
| daf-16 | DA837 | 12.4 ± 2.6 | 234 | 3.0 x 10-45 |
| daf-2 | DA837 | 28.5 ± 9.2 | 90 | 2.1 x 10-16 |
| age-1 | DA837 | 23.7 ± 8.9 | 119 | 4.3 x 10-9 |
| tax-4 | DA837 | 22.0 ± 5.4 | 72 | 3.6 x 10-6 |
| unc-31 | DA837 | 21.2 ± 7.2 | 120 | 3.6 x 10-4 |
| lite-1 | DA837 | 19.7 ± 4.0 | 90 | 0.036 |
| aak-2 | DA837 | 9.6 ± 1.8 | 117 | 3.9 x 10-30 |
| N2 | HT115 (EV RNAi) | 20.9 ± 10.5 | 80 | N/A |
| N2 | HT115 (daf-2 RNAi) | 29.9 ± 10.3 | 38 | 1.2x 10-6 |
| N2 | HT115 (odr-10 RNAi) | 21.5 ± 6.5 | 40 | 0.53 |
| daf-2 | HT115 (EV RNAi) | 30.0 ± 10.5 | 40 | 1.6 x 10-6 |
| daf-2 | HT115 (daf-2 RNAi) | 34.5 ± 9.5 | 39 | 7.9 x 10-11 |
| daf-2 | HT115 (odr-10 RNAi) | 36.9 ± 7.4 | 39 | 8.2 x 10-15 |
| odr-10 | HT115 (EV RNAi) | 21.5 ± 6.6 | 40 | 0.40 |
| odr-10 | HT115 (daf-2 RNAi) | 28.3 ± 12.5 | 38 | 4.8 x 10-4 |
| odr-10 | HT115 (odr-10 RNAi) | 21.1 ± 7.5 | 40 | 0.49 |
Table 2
Summary of paraquat assay survival data.
| Strain | Food source | Lifespan (Mean ± SD) (Days) | N | P-value relative to N2 control |
|---|---|---|---|---|
| N2 | DA837 + 40 mM paraquat | 21.2 ± 8.9 | 58 | N/A |
| daf-16 | DA837 + 40 mM paraquat | 16.5 ± 12.0 | 60 | 0.001 |
| daf-2 | DA837 + 40 mM paraquat | 37.4 ± 22.0 | 68 | 2.0 x 10-5 |
| age-1 | DA837 + 40 mM paraquat | 29.7 ± 13.8 | 68 | 2.7 x 10-5 |
Additional files
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Supplementary file 1
Detailed protocol for WorMotel preparation.
- https://doi.org/10.7554/eLife.26652.027
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Supplementary file 2
Stereolithography (STL) file of the 240-well WorMotel mold.
- https://doi.org/10.7554/eLife.26652.028
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Supplementary file 3
Stereolithography (STL) file of the 48-well WorMotel mold.
- https://doi.org/10.7554/eLife.26652.029
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Supplementary file 4
Parts lists, instructions, and MATLAB software for imaging system and blue light stimulation apparatus.
- https://doi.org/10.7554/eLife.26652.030
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Supplementary file 5
MATLAB software suite for image processing and data analysis, with instructions.
- https://doi.org/10.7554/eLife.26652.031
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Supplementary file 6
Additional MATLAB scripts for data representation.
- https://doi.org/10.7554/eLife.26652.032
