Whole-organism behavioral profiling reveals a role for dopamine in state-dependent motor program coupling in C. elegans
- Picower Institute for Learning & Memory, Department of Brain & Cognitive Sciences, Massachusetts Institute of Technology, United States
Figures
Figure 1 with 3 supplements
Simultaneous measurement of the diverse C. elegans motor programs.
(A) Image of the tracking microscope. (B) Example image of a C. elegans animal from the tracking microscope. Green line denotes detected outline of worm; red line indicates worm’s centerline, …
Figure 1—figure supplement 1
Schematic of the tracking microscope.
An infrared LED trans-illuminates the sample from above via a diffuser and a condenser lens. Light from the sample (red line) is collected by a low-magnification objective underneath the sample, and …
Figure 1—figure supplement 2
Extraction of behavioral parameters from video recordings, and validation of methods.
(A) Movement path of C. elegans animal during recording on tracking microscope. Path is color-coded by velocity (black is slow; red is fast; blue is reverse). (B) Example speed traces over a 6 hr …
Figure 1—figure supplement 3
Event-triggered averages showing behavioral coordination surrounding egg-laying and DMP events.
(A) Event-triggered behavioral averages surrounding egg-laying events. Velocity, angular velocity, feeding rate, and defecation rate are shown as medians ± 25th and 75th percentiles (gray lines …
Figure 2 with 7 supplements
Identifying behavioral states through time series analysis of C. elegans posture.
(A) Schematic showing that body posture is quantified as a vector of relative body angles, from head to tail. (B) Compendium of 100 reference postures that encompass the range of typical C. elegans …
Figure 2—figure supplement 1
Locomotion during the 100 compendium postures.
Average animal velocity across 30 animals during each of the 100 compendium postures. Postures are ordered by posture group. Note that most posture groups (which contain different numbers of …
Figure 2—figure supplement 2
Additional analyses related to Posture-HMM.
(A) A compendium of reference postures can explain most of the variance of observed animal postures. Increasing the number of postures in the compendium leads to a higher percentage of variance …
Figure 2—figure supplement 3
Further characterization of behavioral states from Posture-HMM.
(A) Top: for each state, the most common postures observed (out of the 100 compendium reference postures; Figure 2B) while animals are in the state are shown. The postures exhibited in each of the …
Figure 2—figure supplement 4
Analysis of the dwelling sub-modes.
(A) Example behavioral data from a wild-type animal showing raw posture data and an ethogram indicating the animal’s behavioral state, inferred from the posture-HMM. The color legend for the …
Figure 2—figure supplement 5
The posture-HMM generates posture sequences that resemble those from real animals.
(A) Heat maps showing the differences between synthetic posture sequences and real posture sequences for three different models of postural transitions during dwelling (see Materials and methods for …
Figure 2—figure supplement 6
Locomotion surrounding egg-laying events.
(A) Egg-laying rates over the durations of roaming states. All roaming states were ‘stretched’ so that they could be properly aligned (t = 0 is the onset of the roaming states), and then the …
Figure 2—figure supplement 7
Stereotyped behavioral changes accompany state transitions.
Event-triggered averages are shown for velocity, pumping, and DMPs surrounding the moments of specific types of posture-HMM state transitions, indicated by the rows and columns. Above: we show all …
Figure 3 with 2 supplements
Analysis of neuromodulation mutants reveals a role for dopamine in state-dependent egg-laying.
(A) Behavioral parameters of animals of the indicated genotypes across the posture-HMM states. Black dots indicate significant changes; *p<0.05, Bonferroni-corrected Mann-Whitney U test. The cat-2 …
Figure 3—figure supplement 1
Additional behavioral analysis of mutant strains.
(A) The postures that cat-2 mutant animals display during roaming are similar to those displayed by wild-type. Data are histograms of percent time in each of the eight posture groups during roaming …
Figure 3—figure supplement 2
Full behavioral parameters of the dopamine receptor mutant animals.
Behavioral parameters during each of nine HMM states are shown for animals of the indicated genotypes. Black dots indicate significant changes; *p<0.05, Bonferroni-corrected Mann-Whitney U test. All …
Figure 4 with 1 supplement
The coupling between egg-laying and roaming states leads to greater dispersal of eggs along a food source.
(A) Average distance between a given egg and its k nearest eggs for wild-type animals, separated out for eggs laid while roaming and dwelling. *p<0.01, roaming versus dwelling for all k > 3, …
Figure 4—figure supplement 1
Fraction of eggs laid per state in dopamine pathway mutants.
Quantification of the fraction of eggs that were laid in each of the nine HMM states. Note that this measurement is distinct from egg-laying rate in each state, since this metric also takes into …
Figure 5 with 1 supplement
Optogenetic control of dopaminergic neurons alters egg-laying.
(A) Inhibition of dopaminergic neurons via activation of dat-1::GtACR2 reduces egg-laying rates. Data are shown as egg-laying rates, normalized to pre-stimulation baseline rates. Gray lines show …
Figure 5—figure supplement 1
Additional optogenetic studies and analysis.
(A) No effect of laser illumination on egg-laying rates in the absence of all-trans retinal (ATR) co-factor for dat-1::GtACR2 animals. n = 12 animals. (B) Average velocity during light exposure for …
Figure 6 with 1 supplement
Dopaminergic PDE neurons display activity patterns phase-locked to egg-laying during roaming.
(A) PDE dynamics increase with animal speed. PDE dynamics here is defined as the absolute value of the time derivative of the PDE GCaMP signal. **p<0.001, empirical bootstrap test. (B) Example …
Figure 6—figure supplement 1
Additional analyses related to PDE activity and egg-laying.
(A) Dynamics of ADE, CEPD, and CEPV do not vary with animal speed. Dynamics here is defined as the absolute value of the time derivative of the GCaMP signal for each neuron. n = 22–28 animals per …
Figure 7
Dopamine elevates egg-laying in a GABA-dependent manner.
(A) Effects of dat-1::CoChR activation in genetic mutants with disrupted components of egg-laying circuitry. Data are shown as the fold increase in egg-laying during lights-on period, compared to …
Videos
Video 1
Examples of behavioral states captured through posture-HMM.
This video shows examples of the nine different behavioral states identified through posture-HMM. Note that videos are of different durations and are looped.
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Strain, strainbackground (E. coli) | OP50 | CGC | ID_FlavellDatabase:OP50 | OP50 |
| Strain, strainbackground (C. elegans) | MT13113 | CGC | ID_FlavellDatabase: MT13113 | tdc-1(n3419) |
| Strain, strainbackground (C. elegans) | MT15620 | CGC | ID_FlavellDatabase: MT15620 | cat-2(n4547) |
| Strain, strainbackground (C. elegans) | MT9455 | CGC | ID_FlavellDatabase: MT9455 | tbh-1(n3247) |
| Strain, strainbackground (C. elegans) | CX11078 | Stern et al., 2017 | ID_FlavellDatabase: CX11078 | cat-2(e1112) |
| Strain, strainbackground (C. elegans) | CX14295 | CGC | ID_FlavellDatabase: CX14295 | pdfr-1(ok3425) |
| Strain, strainbackground (C. elegans) | LX645 | CGC | ID_FlavellDatabase: LX645 | dop-1(vs100) |
| Strain, strainbackground (C. elegans) | LX702 | CGC | ID_FlavellDatabase: LX702 | dop-2(vs105) |
| Strain, strainbackground (C. elegans) | LX703 | CGC | ID_FlavellDatabase: LX703 | dop-3 (vs106) |
| Strain, strainbackground (C. elegans) | LX704 | CGC | ID_FlavellDatabase: LX704 | dop-2(vs105); dop-3(vs106) |
| Strain, strainbackground (C. elegans) | SWF261 | this study | ID_FlavellDatabase: SWF261 | dop-4(ok1321); backcrossed to N2 8x |
| Strain, strainbackground (C. elegans) | CX13111 | this study | ID_FlavellDatabase: CX13111 | dop-5(ok568); backcrossed to N2 3x |
| Strain, strainbackground (C. elegans) | RB1680 | CGC | ID_FlavellDatabase: RB1680 | dop-6(ok2070) |
| Strain, strainbackground (C. elegans) | MT1082 | CGC | ID_FlavellDatabase: MT1082 | egl-1(n487) |
| Strain, strainbackground (C. elegans) | SWF266 | this study | ID_FlavellDatabase: SWF266 | lgc-53(n4330); MT13952 was backcrossed to N2 4x |
| Strain, strainbackground (C. elegans) | SWF181 | this study | ID_FlavellDatabase: SWF181 | cat-2(n4547), flvEx87[cat-2 genomic PCR product, myo-3::mCherry] |
| Strain, strainbackground (C. elegans) | SWF325 | this study | ID_FlavellDatabase: SWF325 | flvEx133[dat-1::GtACR2-t2a-GFP,myo-3::mCherry] |
| Strain, strainbackground (C. elegans) | SWF141 | this study | ID_FlavellDatabase: SWF141 | flvEx74[dat-1::CoChR, myo-3::mCherry] |
| Strain, strainbackground (C. elegans) | SWF207 | this study | ID_FlavellDatabase: SWF207 | egl-1(n487); flvEx74[dat-1::CoChR, myo-3::mCherry] |
| Strain, strainbackground (C. elegans) | SWF208 | this study | ID_FlavellDatabase: SWF208 | lin-39(n709); flvEx74[dat-1::CoChR, myo-3::mCherry] |
| Strain, strainbackground (C. elegans) | SWF258 | this study | ID_FlavellDatabase: SWF258 | acr-2(n2595 n2420); flvEx74[dat-1::CoChR, myo-3::mCherry] |
| Strain, strainbackground (C. elegans) | SWF257 | this study | ID_FlavellDatabase: SWF257 | unc-25(e156); flvEx74[dat-1::CoChR, myo-3::mCherry] |
| Strain, strainbackground (C. elegans) | SWF314 | this study | ID_FlavellDatabase: SWF314 | unc-30(e191); flvEx74[dat-1::CoChR, myo-3::mCherry] |
| Strain, strainbackground (C. elegans) | SWF331 | this study | ID_FlavellDatabase: SWF331 | flvEx127[dat-1::GCaMP6m, myo-3::mCherry] |
| Strain, strainbackground (C. elegans) | BZ555 | CGC | ID_FlavellDatabase: BZ555 | egIs1 [dat-1::GFP] |
| Strain, strainbackground (C. elegans) | CX14453 | Bendesky et al., 2012 | ID_FlavellDatabase: CX14453 | unc-25(n2324) |
| Strain, strainbackground (C. elegans) | CX13851 | Bendesky et al., 2012 | ID_FlavellDatabase: CX13851 | unc-25(e156) |
| Recombinant DNA reagent | pYCH1 | this study | ID_FlavellDatabase: pYCH1 | dat-1::GtACR2-sl2-GFP |
| Recombinant DNA reagent | pSKY1 | this study | ID_FlavellDatabase: pSKY1 | dat-1::CoChR |
| Recombinant DNA reagent | pSKY2 | this study | ID_FlavellDatabase: pSKY2 | dat-1::GCaMP6m |
| Software, algorithm | ImageJ | ImageJ (http://imagej.nih.gov/ij/) | RRID:SCR_003070 | Version 1.52 |
| Software, algorithm | GraphPad Prism | GraphPad Prism (graphpad.com) | RRID:SCR_002798 | Version 7.03 |
| Software, algorithm | MATLAB | MathWorks (www.mathworks.com) | RRID:SCR_001622 | Version 2019a |
| Software, algorithm | National Instruments | LabView (www.ni.com/en-us/shop/labview.html) | RRID:SCR_014325 | Version 16.0 |
| Software, algorithm | NIS Elements | Nikon (www.nikoninstruments.com/products/software) | RRID:SCR_014329 | V4.51.01 |
| Software, algorithm | The R Project | R (r-project.org) | RRID:SCR_001905 | v3.6.1 |
| Software, algorithm | R Studio | R Studio (rstudio.com) | RRID:SCR_000432 | v1.2.1335 |
Additional files
-
Supplementary file 1
Details about statistical tests.
This excel sheet includes a detailed description of each statistical test carried out in this study.
- https://cdn.elifesciences.org/articles/57093/elife-57093-supp1-v2.xlsx
-
Transparent reporting form
- https://cdn.elifesciences.org/articles/57093/elife-57093-transrepform-v2.pdf
