Sensory neurons couple arousal and foraging decisions in Caenorhabditis elegans
- Lulu and Anthony Wang Laboratory of Neural Circuits and Behavior, The Rockefeller University, United States
Figures
Figure 1 with 2 supplements
Animals make foraging decisions at the boundaries of bacterial food lawns.
(A) Image of C. elegans on a small lawn of bacteria. Head is indicated with a green dot, lawn boundary is indicated with a white line. Scale bar is 1 mm. (B) Schematic depicting lawn boundary …
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Figure 1—source data 1
Quantification of various lawn boundary interaction behaviors from experiments in Figure 1.
- https://cdn.elifesciences.org/articles/88657/elife-88657-fig1-data1-v1.xlsx
Figure 1—figure supplement 1
Small bacterial lawns are denser at the lawn boundary, and quantification of duration outside the lawn (A) A small lawn of bacteria expressing green fluorescent protein (GFP) imaged in the GFP channel.
White scale bar is 1 mm. (B) GFP intensity across the orange transect line plotted in (A) shows accumulation of bacterial cells near the lawn boundary in brighter GFP pixels. (C) An image of the …
Figure 1—video 1
Lawn boundary encounters lead to different behavioral responses: lawn leaving, head poke forward, head poke pause, and head poke reversal.
Green dot indicates the head position. White line indicates the lawn boundary. Forward and Reverse motion indicated in top left corner.
Figure 2 with 6 supplements
Lawn leaving is associated with high arousal states on multiple timescales.
(A) Midbody forward speed aligned to lawn leaving. Left, heatmap of individual speed traces. Right, mean midbody forward speed computed across the heatmap traces. White space indicates missing data …
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Figure 2—source data 1
Quantification of behavioral states surrounding lawn leaving events from experiments in Figure 2.
- https://cdn.elifesciences.org/articles/88657/elife-88657-fig2-data1-v1.xlsx
Figure 2—figure supplement 1
Roaming and dwelling states on small bacterial lawns.
(A) Scatter plot of average absolute speed and angular speed in 10 s intervals for wild type animals on small lawns (n=1586 animals, 380,640 bins). The boundary shown, used for all data in the …
Figure 2—figure supplement 2
Modeling behavioral states across locomotory feature dimensions using an Autoregressive Hidden Markov Model.
(A) Schematic of animal body shape with key body points indicated. (B–E) Schematics illustrating derived behavioral features. See Methods for detailed description. (B) Midbody speed, (C) Midbody …
Figure 2—figure supplement 3
Head Poke Reversals are associated with small changes in arousal states.
(A) Midbody forward speed aligned to head poke reversals. Left, heatmap of individual speed traces. Right, mean midbody forward speed computed across the heatmap traces. (B) Overlap of head poke …
Figure 2—figure supplement 4
Lawn boundary distance distributions by HMM state (A–D) Empirical distributions of lawn boundary distance by HMM state.
Positive values indicate distances inside the lawn, negative values indicate distances outside the lawn (See Figure 1). (A) Lawn boundary distance of animals while roaming and dwelling. Statistics …
Figure 2—video 1
Example of animal behavior with roaming and dwelling annotated.
Animal shown corresponds to the behavior before and after the first lawn leaving event in Figure 2—figure supplement 2P.
Green dot indicates the head position. White line indicates the lawn boundary. Forward and Reverse motion indicated in top left corner. Speed indicated top middle. Time stamps indicated top right. Video sped up 7 x.
Figure 2—video 2
The same animal and time steps as in Figure 2—video 1, except annotated with AR-HMM states instead of roaming and dwelling.
Figure 3 with 1 supplement
Food intake regulates arousal states and lawn leaving.
(A–B) Increasing bacterial density suppresses roaming and leaving. (A) Fraction of time roaming on small lawns seeded with bacteria of different optical density (OD). OD1: n=45, OD2: n=46, OD4: …
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Figure 3—source data 1
Quantification of roaming and lawn leaving from experiments in Figure 3.
- https://cdn.elifesciences.org/articles/88657/elife-88657-fig3-data1-v1.xlsx
Figure 3—figure supplement 1
Further quantification of roaming and leaving behaviors under food intake inhibition.
(A–B) Aztreonam affects roaming and leaving by affecting bacterial growth. In ‘pre-add’ conditions, the drug is added to liquid cultures and agar plates during bacterial growth. In ‘post-add’ …
Figure 4 with 1 supplement
Neuromodulatory signaling mutants retain coupling of arousal and leaving.
(A–P) Roaming, lawn leaving, roaming before leaving, and roaming speed quantified across mutants in four neuromodulatory genes known to alter roaming and dwelling: tph-1, cat-2, npr-1, and pdfr-1. (A…
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Figure 4—source data 1
Quantification of roaming and lawn leaving from experiments in Figure 4.
- https://cdn.elifesciences.org/articles/88657/elife-88657-fig4-data1-v1.xlsx
Figure 4—figure supplement 1
Optogenetic feeding inhibition stimulates roaming and leaving in neuromodulatory mutants.
(A–B) Optogenetic feeding inhibition in pdfr-1 mutants. (A) Left, Fraction of animals roaming before, during and after optogenetic feeding inhibition in pdfr-1 mutants and paired wild type controls. …
Figure 5 with 3 supplements
Acute circuit manipulation drives deterministic roaming and probabilistic leaving.
(A) Experimental design. Stimulation of bacterial light-activated adenylyl cyclase (BlaC) with 455 nm light increases cAMP synthesis in RIB::BlaC neurons. (B) Heatmap showing roaming and dwelling …
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Figure 5—source data 1
Quantification of roaming and lawn leaving from experiments in Figure 5.
- https://cdn.elifesciences.org/articles/88657/elife-88657-fig5-data1-v1.xlsx
Figure 5—figure supplement 1
pdfr-1 genomic characteristics and expression patterns.
(A) Genomic region surrounding pdfr-1 (black boxes represent coding exons, gray boxes are non-coding exons), region deleted in pdfr-1(ok3425) mutants, proximal and distal promoters, PCR fragments …
Figure 5—figure supplement 2
Transgenic rescue of pdfr-1 in RIB neurons restores roaming (A) Schematic depicting intersectional cell-specific rescue of pdfr-1 using an inverted Cre-Lox strategy.
(B) Restoring pdfr-1 expression in AIY, RIM, and RIA neurons did not rescue roaming on small lawns (wild type n=64, inverted pdfr-1 transgene n=64, AIY, RIM, RIA pdfr-1 rescue n=64, …
Figure 5—figure supplement 3
Further quantification and controls of RIB::BlaC experiments.
(A–E) Quantification of RIB::BlaC controls. (A) Blue light stimulation does not induce roaming in wild-type animals. Statistics by paired t-test on logit-transformed data. (B) Blue light stimulation …
Figure 6 with 4 supplements
tax-4-expressing sensory neurons couple roaming and lawn leaving.
(A–C) Roaming and leaving in tax-4 mutants, and rescue by tax-4 expression in ASJ neurons (wild type n=143, tax-4 n=156, tax-4 ASJ rescue n=148). Additional features of roaming and leaving are shown …
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Figure 6—source data 1
Quantification of roaming and lawn leaving from experiments in Figure 6.
- https://cdn.elifesciences.org/articles/88657/elife-88657-fig6-data1-v1.xlsx
Figure 6—figure supplement 1
Further quantification of tax-4 mutants and tax-4 ASJ rescue on edible food (A) Complementary cumulative distribution functions (ccdfs) for overall roam state durations for wild type, tax-4, and tax-4 rescued in ASJ.
Different letters mark significant differences by Kolmogorov-Smirnov two-sample tests with Bonferroni correction. (B) Same as (A) for roam state durations before leaving events. Wild type n=143, tax-…
Figure 6—figure supplement 2
tax-4 rescue in additional tax-4-expressing neurons (A–B) Quantification of roaming and leaving in tax-4 mutants with tax-4 rescued in ASJ, ASK or ASJ +ASK neurons (wild type n=95, tax-4 n=104, ASJ rescue n=91, ASK rescue n=101, ASJ +ASK rescue n=105).
(A) ASJ or ASJ +ASK tax-4 rescue restores roaming to tax-4 mutants. (B) ASK tax-4 rescue suppresses lawn leaving below tax-4 levels. (C–D) Quantification of roaming and leaving in tax-4 mutants with …
Figure 6—figure supplement 3
Further quantification of tax-4 mutants and tax-4 rescue upon optogenetic feeding inhibition (A-D) Effects of tax-4 on roaming and leaving behavior induced by optogenetic feeding inhibition.
(A) Fraction of animals roaming before, during, and after optogenetic feeding inhibition. Light ON period denoted by yellow shading. (B) Cumulative distribution of time until the first head poke …
Figure 6—figure supplement 4
tax-4 mutants leave less on inedible food and ASJ rescue restores leaving.
(A–B) In these panels, wild type n=30, tax-4 n=22, tax-4 ASJ rescue n=10 on inedible food generated by pre-treatment of bacterial lawns with aztreonam before growth. (A) Wild type, tax-4 and tax-4 …
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Strain, strain background (Caenorhabditis elegans, N2 hermaphrodite) | Wild type | Yoshimura et al., 2019 | PD1074 (now CGC1) | Used in all figures except Figure 4 tph-1 and cat-2 controls |
| Strain, strain background (Caenorhabditis elegans, N2 hermaphrodite) | Wild type | This paper | ID_BargmannDatabase:CX0001 | Used as wild type controls in Figure 4 tph-1 and cat-2 experiments |
| Strain, strain background (Caenorhabditis elegans, N2 hermaphrodite) | pharynx ReaChR | This paper | ID_BargmannDatabase:CX16279 | Figures 3 and 6 and supplements |
| Strain, strain background (Caenorhabditis elegans, N2 hermaphrodite) | pdfr-1 | Flavell et al., 2013 | ID_BargmannDatabase:CX14295 | Figure 4 |
| Strain, strain background (Caenorhabditis elegans, N2 hermaphrodite) | tph-1 | https://cgc.umn.edu/strain/MT15434 | ID_BargmannDatabase:MT15434 | Figure 4 |
| Strain, strain background (Caenorhabditis elegans, N2 hermaphrodite) | cat-2 | Stern et al., 2017 | ID_BargmannDatabase:CX11078 | Figure 4 |
| Strain, strain background (Caenorhabditis elegans, N2 hermaphrodite) | npr-1 | Jang et al., 2017 | ID_BargmannDatabase:CX13663 | Figure 4 |
| Strain, strain background (Caenorhabditis elegans, N2 hermaphrodite) | pharynx ReaChR; pdfr-1 | This paper | ID_BargmannDatabase:CX16528 | Figure 4—figure supplement 1 |
| Strain, strain background (Caenorhabditis elegans, N2 hermaphrodite) | pharynx ReaChR; tph-1 | This paper | ID_BargmannDatabase:CX16529 | Figure 4—figure supplement 1 |
| Strain, strain background (Caenorhabditis elegans, N2 hermaphrodite) | RIB::BlaC | This paper | ID_BargmannDatabase:CX18471 | Figure 5, Figure 5—figure supplement 3 |
| strain, strain background (Caenorhabditis elegans, N2 hermaphrodite) | PCR fragment #1; pdfr-1 | Flavell et al., 2013 | ID_BargmannDatabase:CX14378 | Figure 5—figure supplement 1 |
| Strain, strain background (Caenorhabditis elegans, N2 hermaphrodite) | PCR fragment #2; pdfr-1 | Flavell et al., 2013 | ID_BargmannDatabase:CX14383 | Figure 5—figure supplement 1 |
| Strain, strain background (Caenorhabditis elegans, N2 hermaphrodite) | pdfr-1:: CreONpdfr-1 | Flavell et al., 2013 | ID_BargmannDatabase:CX14485 | Figure 5—figure supplement 2 |
| Strain, strain background (Caenorhabditis elegans, N2 hermaphrodite) | RIB pdfr-1 rescue; pdfr-1 | This paper | ID_BargmannDatabase:CX18302 | Figure 5—figure supplement 2 |
| Strain, strain background (Caenorhabditis elegans, N2 hermaphrodite) | pan-neuronal pdfr-1 rescue; pdfr-1 | Flavell et al., 2013 | ID_BargmannDatabase:CX14488 | Figure 5—figure supplement 2 |
| Strain, strain background (Caenorhabditis elegans, N2 hermaphrodite) | AIY, RIM, RIA pdfr-1 rescue; pdfr-1 | Flavell et al., 2013 | ID_BargmannDatabase:CX14271 | Figure 5—figure supplement 2 |
| Strain, strain background (Caenorhabditis elegans, N2 hermaphrodite) | tax-4 | Worthy et al., 2018 | ID_BargmannDatabase:CX13078 | Figure 6, Figure 6—figure supplements 1 and 2, 4 |
| Strain, strain background (Caenorhabditis elegans, N2 hermaphrodite) | ASJ rescue; tax-4 | This paper | ID_BargmannDatabase:CX11118 | Figure 6, Figure 6—figure supplements 2 and 4 |
| Strain, strain background (Caenorhabditis elegans, N2 hermaphrodite) | ASK rescue; tax-4 | This paper | ID_BargmannDatabase:CX13361 | Figure 6—figure supplement 2 |
| Strain, strain background (Caenorhabditis elegans, N2 hermaphrodite) | ASJ +ASK rescue; tax-4 | This paper | ID_BargmannDatabase:CX11110 | Figure 6—figure supplement 2 |
| Strain, strain background (Caenorhabditis elegans, N2 hermaphrodite) | AWC rescue; tax-4 | Worthy et al., 2018 | ID_BargmannDatabase:CX13790 | Figure 6—figure supplement 2 |
| Strain, strain background (Caenorhabditis elegans, N2 hermaphrodite) | ASI rescue; tax-4 | This paper | ID_BargmannDatabase:CX11558 | Figure 6—figure supplement 2 |
| Strain, strain background (Caenorhabditis elegans, N2 hermaphrodite) | URX/AQR/PQR rescue; tax-4 | This paper | ID_BargmannDatabase:CX11113 | Figure 6—figure supplement 2 |
| Strain, strain background (Caenorhabditis elegans, N2 hermaphrodite) | pharynx ReaChR; tax-4 | This paper | ID_BargmannDatabase:CX18452 | Figure 6, Figure 6—figure supplement 3 |
| Strain, strain background (Caenorhabditis elegans, N2 hermaphrodite) | pharynx ReaChR; tax-4; ASJ rescue | This paper | ID_BargmannDatabase:CX18538 | Figure 6, Figure 6—figure supplement 3 |
| Strain, strain background (Escherichia coli, OP50) | OP50 | Caenorhabditis Genetics Center (CGC) | https://cgc.umn.edu/strain/OP50 | |
| Chemical compound, drug | Aztreonam | Sigma | PZ0038 | CAS: 78110-38-0 |
| Chemical compound, drug | all trans-Retinal (ATR) | Sigma | R2500 | CAS: 116-31-4 |
| Software, algorithm | ImageJ | ImageJ (https://imagej.nih.gov/) | RRID:SCR_003070 | Version 1.50i |
| Software, algorithm | MATLAB | Mathworks (https://www.mathworks.com/) | RRID:SCR_001622 | Version R2018a, R2020a, R2021a, R2023a |
| Software, algorithm | FlyCapture | Pointgrey (https://www.ptgrey.com/) | Version FlyCap2 | |
| Software, algorithm | Python | Python (python.org) | RRID:SCR_008394 | Version 3.8.3 |
| Software, algorithm | tracking and analysis code | this paper; Scheer and Bargmann, 2023 | https://github.com/BargmannLab/Scheer_Bargmann2023 |
Additional files
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Supplementary file 1
Sensory neurons and endocrine systems implicated in roaming and leaving behavior.
- https://cdn.elifesciences.org/articles/88657/elife-88657-supp1-v1.xlsx
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Supplementary file 2
Lawn Leaving Events Per minute Roaming/Dwelling.
- https://cdn.elifesciences.org/articles/88657/elife-88657-supp2-v1.xlsx
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Supplementary file 3
Strains and Plasmids.
- https://cdn.elifesciences.org/articles/88657/elife-88657-supp3-v1.xlsx
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Supplementary file 4
p-values and Statistical Tests.
- https://cdn.elifesciences.org/articles/88657/elife-88657-supp4-v1.xlsx
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MDAR checklist
- https://cdn.elifesciences.org/articles/88657/elife-88657-mdarchecklist1-v1.pdf
