Article Figures & Data
Figures
- Figure 1.
GLR-1 abundance declines in wild-type worms during aging. A, Age-dependent chemotaxis decline toward DA in wild-type worms. B, Nose touch response of young, 2-d-old, and 3-d-old wild-type adult worms. C, Scheme illustrating the location for in vivo imaging of GLR-1 protein intensities and dynamics in the AVA neuron. D, The GLR-1 receptor abundance was measured using confocal microscopy in transgenic worms in the wild-type background. Representative image of the membrane-bound and total GLR-1 content of the AVA axon in young (left) and 3-d-old (right) wild-type animals. E–H, Box plots displaying quantification of the fluorescence intensity of (E) total (mCherry) and (F) membrane-bound (SEP) GLR-1 receptors normalized to Day 1 signal in the proximal part of the axon of the AVA neuron; (G) total (mCherry) and (H) membrane-bound (SEP) GLR-1 level in the distal parts of AVA axon of SEP::mCherry::GLR-1(wt) worms. I, J, Membrane-bound versus total GLR-1 ratio in the proximal (I) and distal part (J) of AVA axon in 1-d-old (left) and 3-d-old (right) wild-type animals; (K) glr-1 RNA levels were measured with real-time qRT-PCR in 1- and 3-d-old SEP::mCherry::GLR-1(WT) strain. Expression levels were normalized to tba-1 and cdc-42. L, Fluorescence intensity levels of the mCherry (total GLR-1) signal in young and aged SEP::mCherry::GLR-1(WT) animals in the cell body of the AVA neuron. Scale bars, 5 µm. Data on (A) and (E–L) are visualized with Tukey's boxplots. See Extended Data Table 2 for detailed statistical information.
- Figure 2.
GLR-1 transport does not decline in wild-type worms at Day 3. A, D, Time-lapse images of GLR-1 vesicle movements along the axon of the AVA neuron. The moving vesicle is highlighted with arrowheads. The bottom panel shows a kymogram of all merged timeframes where transport events can be seen as oblique lines crossing. A, Anterograde transport from the soma (vesicle; blue arrowhead). D, Representative example of the retrograde transport (vesicle; red arrowhead). B, Average number of anterograde and (C) retrograde transport events in the proximal part of AVA. E, Average number of anterograde and (F) retrograde transport events in the distal part of AVA of SEP::mCherry::GLR-1(wt) animals. Scale bars, 5 µm. Data on B, C, E, and F are visualized with Tukey's boxplots. See Extended Data Table 2 for detailed statistical information.
- Figure 3.
Age-dependent GLR-1 dynamics decline in wild-type worms. A, B, Representative image of the FRAP recovery of the membrane-bound (A) and total GLR-1 (B) content of the AVA axon in young and aged wild-type animals over time. C, E, Percentage FRAP recovery in the proximal part for (C) mCherry fluorescence over time at Day 1 (plateau, 0.5136) and at Day 3 (plateau, 0.3718) and (E) SEP fluorescence at Day 1 (plateau, 0.4395) and at Day 3 (plateau, 0.3428; n > 22 for each group). D, F, Quantity of the mobile fraction of (D) total GLR-1 and (F) membrane-bound GLR-1 during aging in the proximal part of the AVA neuron. G, I, Percent recovery of (G) mCherry fluorescence after photobleaching over time at Day 1 (plateau, 0.34) and at Day 3 (plateau, 0.2250) and (I) SEP fluorescence at Day 1 (plateau, 0.3555) and at Day 3 (plateau, 0.1983; n > 20 for each group) in the distal part. H, J, Relative quantity of the mobile fraction of (H) total GLR-1 and (J) membrane-bound GLR-1 during aging in the distal part of the AVA neuron during aging. Scale bar, 5 µm. Data on D, F, H, and J are visualized with Tukey's boxplots, and C, E, G, and I show mean ± SEM. See Extended Data Table 2 for detailed statistical information.
- Figure 4.
GLR-1 abundance is maintained with age in ubiquitination-defective mutants. A, B, Box plots displaying quantification of the fluorescence intensity of (A) total GLR-1 and (B) membrane-bound receptors normalized to Day 1 signal in the proximal part of the axon of AVA neuron of 4KR mutant worms. C,D, Receptor levels of (C) total GLR-1 and (D) membrane bound in the distal part of AVA axon of 4KR mutant worms. E,F, Membrane-bound versus total GLR-1 ratio in the proximal (E) and distal part (F) of the AVA axon in 1-d-old (left) and 3-d-old (right) animals carrying a ubiquitination-defective GLR-1 reporter. Data on A–F are visualized with Tukey's boxplots. See Extended Data Table 2 for detailed statistical information.
- Figure 5.
GLR-1 dynamics is maintained with age in ubiquitination-defective mutants. A, C, Percent fluorescence recovery over time after photobleaching of (A) mCherry at Day 1 (plateau, 0.4062) and at Day 3 (plateau, 0.4493) and (C) SEP::GFP at Day 1 (plateau, 0.3358) and at Day 3 (plateau, 0.3728; n > 13 for each group) in the proximal part of AVA neuron. B, D, Mobile fraction of (B) total GLR-1 and (D) membrane-bound GLR-1 during aging in the proximal part of the AVA neuron. E, G, Percent fluorescence recovery of (E) mCherry after photobleaching over time at Day 1 (plateau, 0.4079) and at Day 3 (plateau, 0.3024) and (G) SEP at Day 1 (plateau, 0.2801) and at Day 3 (plateau, 0.2632; n ≥ 20 for each group) in the distal part. F, H, Quantification of the mobile fraction of (F) total GLR-1 and (H) membrane-bound GLR-1 in the distal part of the AVA neuron during aging. Data on B, D, F, and H are visualized with Tukey's boxplots. Panels A, C, E, and G show mean ± SEM. See Extended Data Table 2 for detailed statistical information.
- Figure 6.
GLR-1 abundance remains unchanged in msi-1(lf) mutants with aging. A, B, Quantification of the fluorescence intensity of (A) total and (B) membrane-bound GLR-1 receptors normalized to Day 1 signal in the proximal part of the axon of AVA neuron of msi-1(lf) mutant worms. C, D, Receptor levels of (C) total GLR-1 and (D) membrane bound in the distal part of AVA axon. E, F, mCherry fluorescence intensity was compared at Day 1 between SEP::mCherry::GLR-1(WT) and msi-1(lf) mutant animals in the (E) proximal and (F) distal part of the AVA axon. G, Life span of wild-type and msi-1(lf) mutant animals. Data on A–F are visualized with Tukey's boxplots. See Extended Data Table 2 for detailed statistical information.
- Figure 7.
GLR-1 dynamics remain unchanged in msi-1(lf) mutants with aging. A, C, Fluorescence recovery following photobleaching of (A) mCherry at Day 1 (plateau, 0.4681) and at Day 3 (plateau, 0.4793) and (C) SEP at Day 1 (plateau, 0.4341) and at Day 3 (plateau, 0.4562; n > 16 for each group) in the proximal part of AVA neuron. B, D, Box plots displaying mobile fraction of (B) total GLR-1 and (D) membrane-bound GLR-1 during aging in the proximal part of the AVA neuron. E, G, Percent fluorescence recovery of (E) mCherry after photobleaching over time at Day 1 (plateau, 0.3988) and at Day 3 (plateau, 0.3932) and (G) SEP::GFP at Day 1 (plateau, 0.37) and at Day 3 (plateau, 0.3240; n > 17 for each group) in the distal part. F, H, Relative quantity of the mobile fraction of (F) total GLR-1 and (H) membrane-bound GLR-1 during aging in the distal part of the AVA neuron of msi-1(lf) mutants at Day 1 and Day 3. Data on B, D, F, and H are visualized with Tukey's boxplots. Panels A, C, E, and G show mean ± SEM. See Extended Data Table 2 for detailed statistical information.
- Figure 8.
Memory is maintained in GLR-1(4KR) mutants with age. A, B, Short-term aversive olfactory memory assay of wild-type, transgenic animals carrying either SEP::mCherry::GLR-1(wt) or SEP::mCherry::GLR-1(4KR) using DA was tested in (A) young adult and (B) 3-d-old worms. C, Learning index [(CINaive − CIconditioning) / CINaive] or (D) memory index [(CINaive − CI1h delay) / CINaive] was used to normalize for the age-dependent decrease in chemotaxis. E, Short-term aversive olfactory memory assay of wild-type and msi-1(lf) mutant worms was tested in young adults and 3-d-old worms as indicated. F, Nose touch response of young, 2-d-old, and 3-d-old wild-type and transgenic animals carrying either SEP::mCherry::GLR-1(wt) or SEP::mCherry::GLR-1(4KR). Data are visualized with Tukey's boxplots. See Extended Data Table 2 for detailed statistical information.
Extended Data
Extended Data Table 1
Summary table of the primers used for quantitative real-time PCR. Download Extended Data Table 1, XLS file.
Extended Data Table 2
Detailed description of the statistical data for figures 1-8. Download Extended Data Table 2, XLS file.
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