Reliability of an interneuron response depends on an integrated sensory state
- The Rockefeller University, United States
- Chan Zuckerberg Initiative, United States
Figures
Figure 1 with 3 supplements
Optogenetic activation of AWA sensory neurons elicits unreliable AIA calcium responses.
(A and B) AWA GCaMP2.2b (A) or AIA GCaMP5A (B) calcium responses to 10 s pulses of increasing concentrations of diacetyl and to AWA optogenetic stimulation. Bold lines indicate mean response, and …
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Figure 1—source data 1
Source data for Figure 1 and figure supplements.
- https://cdn.elifesciences.org/articles/50566/elife-50566-fig1-data1-v2.xlsx
Figure 1—figure supplement 1
Experimental configuration and calibration of simultaneous GCaMP-Chrimson imaging conditions.
(A) Schematic of experimental configuration. Animals are paralyzed in a microfluidic device and their neural activity is recorded during exposure to odor or light. Two arenas can be recorded …
Figure 1—figure supplement 2
Validation of AIA response thresholding procedure.
(A) Proportion of AIA calcium traces counted as ‘responses’ at varying fluorescence (x-axis) and time derivative (color axis) threshold parameters to various stimuli. Thresholds depend on the …
Figure 1—figure supplement 3
Sequential imaging of AIA responses to odor or AWA::Chrimson stimulation.
(A) Heat maps of AWA and AIA responses to AWA::Chrimson stimulation, combined over all experiments. AWA: n = 268; AIA: n = 569. (B) Delay between the time at which 50% of AWA versus 50% of AIA …
Figure 2 with 1 supplement
Gap junctions mediate AWA-to-AIA communication.
(A, C, D and E) Cumulative response time profiles of AIA responses to 1.15 µM diacetyl in WT versus odr-7 animals (AWA cell fate mutants) (A), odr-10 animals (AWA diacetyl receptor mutants) (A), …
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Figure 2—source data 1
Source data for Figure 2 and figure supplement.
- https://cdn.elifesciences.org/articles/50566/elife-50566-fig2-data1-v2.xlsx
Figure 2—figure supplement 1
Gap junctions contribute to AWA-to-AIA communication, additional data.
(A) Cumulative response time profiles of AIA responses to AWA::Chrimson stimulation in WT versus animals expressing Tetanus Toxin Light Chain A (TeTx) in AWA. (B and D) Cumulative response time …
Figure 3 with 1 supplement
Chemical synapses inhibit AIA.
(A) Cumulative response time profiles of AIA responses to 11.5 nM diacetyl in WT versus unc-13(e51) and unc-18(e234) animals (synaptic transmission mutants). (B) Heat maps of AIA responses to …
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Figure 3—source data 1
Source data for Figure 3 and figure supplement.
- https://cdn.elifesciences.org/articles/50566/elife-50566-fig3-data1-v2.xlsx
Figure 3—figure supplement 1
Chemical synapses inhibit AIA, additional data.
(A) Cumulative response time profiles of AIA to 1.15 µM diacetyl in WT versus unc-13(e51), unc-18(e234), and unc-18(e81) animals (synaptic transmission mutants). (B) Heat maps of AIA responses to …
Figure 4 with 2 supplements
Glutamatergic sensory neurons cooperate to inhibit AIA.
(A) Simplified diagram of connections between AWA, AIA and four glutamatergic sensory neurons, based on White et al. (1986). (B) Schematic of cell-selective glutamate knockout genetic strategy (López…
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Figure 4—source data 1
Source data for Figure 4 and figure supplements.
- https://cdn.elifesciences.org/articles/50566/elife-50566-fig4-data1-v2.xlsx
Figure 4—figure supplement 1
Controls and heat maps for FRT-FLP recombination.
(A) Heat maps of AIA responses to AWA::Chrimson in eat-4-FRT control strain, unc-18, FRT+nFlippase animals lacking glutamate release in specific sensory neurons, shown in Figure 4C–G, WT, and WT …
Figure 4—figure supplement 2
Additional representations of AIA data from Figures 1–4.
(A – C) Negative correlation between AIA GCaMP fluorescence at baseline and magnitude of responses to AWA::Chrimson (A), 11.5 nM diacetyl (B), and 1.15 µM diacetyl (C). Only responses to the first …
Figure 5 with 3 supplements
Multiple sensory neurons detect diacetyl and isoamyl alcohol.
(A – C) Mean ASK (A), AWC (B), and ASE (C) responses to 10 s pulses of buffer (0) or 11.5 nM or 1.15 µM diacetyl. ASK: n = 82–115; AWC: n = 52–60; ASE: n = 42–54. Shading indicates ± SEM. (D) Mean …
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Figure 5—source data 1
Source data for Figure 5 and Figure 5—figure supplements 1–3.
- https://cdn.elifesciences.org/articles/50566/elife-50566-fig5-data1-v2.xlsx
Figure 5—figure supplement 1
Controls for diacetyl activation of sensory neurons.
(A – C) Magnitude of individual ASK (A), AWC (B), or ASE (C) responses to buffer (0), 11.5 nM or 1.15 µM diacetyl shown in Figure 5A–C. Boxes show median and interquartile range. (D) Magnitude of …
Figure 5—figure supplement 2
Simultaneous recording of multiple neurons.
(A and B) Cytoplasmic AIA GCaMP5A and nuclear AWA GCaMP6s and ASK GCaMP6s responses to 10 s pulses of 1.15 µM (A) or 11.5 nM (B) diacetyl, with all three neurons recorded simultaneously in the same …
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Figure 5—figure supplement 2—source data 1
Source data for Figure 5—figure supplement 2.
- https://cdn.elifesciences.org/articles/50566/elife-50566-fig5-figsupp2-data1-v2.xlsx
Figure 5—figure supplement 3
Controls and heat maps for isoamyl alcohol stimulation.
(A – C) Magnitude of individual AWC (A), AWA (B), and ASK (C) responses to buffer (0), 0.9 µM, 9 µM, and 9 µM isoamyl alcohol shown in Figure 5E–G. Boxes show median and interquartile range. (D – F) …
Figure 6 with 1 supplement
AIA neurons are bistable and act as a nonlinear AND-gate.
(A) Representative example of membrane potential dynamics induced by current injection steps in current-clamped AIA neurons (n=45 AIA neurons recorded, all showing bimodal dynamics). Top, current …
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Figure 6—source data 1
Source data for Figure 6A–B and figure supplement.
- https://cdn.elifesciences.org/articles/50566/elife-50566-fig6-data1-v2.zip
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Figure 6—source data 2
Source data for Figure 6C.
- https://cdn.elifesciences.org/articles/50566/elife-50566-fig6-data2-v2.xlsx
Figure 6—figure supplement 1
Current-voltage relationship in AIA.
(A) Representative example of whole-cell current traces induced by a series of voltage steps in voltage-clamped AIA neuron. Top, voltage step protocol: a series of 0.5 s square steps starting at …
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Strain, strain background (Caenorhabditis elegans N2, hermaphrodite) | AWA::GCaMP2.2b | DOI: 10.1016/j.celrep.2015.08.032 | ID_BargmannDatabase:CX14647 | See Figure 1, Figure 1—figure supplements 1, Figure 3, Figure 5—figure supplement 3 |
| Strain, strain background (C. elegans N2, hermaphrodite) | AWA::Chrimson; AWA::GCaMP2.2b | DOI: 10.1016/j.celrep.2015.08.032 | ID_BargmannDatabase:CX16573 | See Figure 1, Figure 1—figure supplements 1, 3, Figure 3, Figure 3—figure supplement 1 |
| Strain, strain background (C. elegans N2, hermaphrodite) | AIA::GCaMP5A | DOI: 10.1016/j.celrep.2015.08.032 | ID_BargmannDatabase:CX15257 | See Figure 1, Figure 1—figure supplements 2, 3, Figure 3—figure supplement 1, Figure 5, Figure 4—figure supplements 2, Figure 5—figure supplement 3, Figure 6, Figure 6—figure supplement 1 |
| Strain, strain background (C. elegans N2, hermaphrodite) | AWA::Chrimson; AIA::GCaMP5A | DOI: 10.1016/j.celrep.2015.08.032 | ID_BargmannDatabase:CX16561 | See Figure 1, Figure 1—figure supplements 2, 3, Figure 2—figure supplement 1, Figure 3, Figure 3—figure supplement 1, Figure 4, Figure 4—figure supplements 1, 2, Figure 5—figure supplement 3 |
| Strain, strain background (C. elegans N2, hermaphrodite) | odr-7; AIA::GCaMP5A | DOI: 10.1016/j.celrep.2015.08.032 | ID_BargmannDatabase:CX16171 | See Figure 2, Figure 5, Figure 4—figure supplement 2, Figure 5—figure supplement 3 |
| Strain, strain background (C. elegans N2, hermaphrodite) | odr-10; AIA::GCaMP5A | DOI: 10.1016/j.celrep.2015.08.032 | ID_BargmannDatabase:CX16170 | See Figure 2, Figure 4—figure supplement 2 |
| Strain, strain background (C. elegans N2, hermaphrodite) | AWA::TeTx; AIA::GCaMP5A | this paper | ID_BargmannDatabase:CX16584 | See Figure 2, Figure 4—figure supplement 2 |
| Strain, strain background (C. elegans N2, hermaphrodite) | AWA::TeTx; AWA::Chrimson; AIA::GCaMP5A | this paper | ID_BargmannDatabase:CX17519 | See Figure 2—figure supplement 1, Figure 4—figure supplement 2 |
| Strain, strain background (C. elegans N2, hermaphrodite) | unc-7(e5); AIA::GCaMP5A | this paper | ID_Bargmann Database:CX18039 | See Figure 2, Figure 4—figure supplement 2 |
| Strain, strain background (C. elegans N2, hermaphrodite) | unc-9(fc16); AIA::GCaMP5A | this paper | ID_BargmannDatabase:CX16980 | See Figure 2 |
| Strain, strain background (C. elegans N2, hermaphrodite) | unc-9 unc-7; AIA::GCaMP5A | this paper | ID_BargmannDatabase:CX16979 | See Figure 2, Figure 2—figure supplement 1, Figure 4—figure supplement 2 |
| Strain, strain background (C. elegans N2, hermaphrodite) | unc-9 unc-7; AWA,AIA::unc-9(-); AIA::GCaMP5A | this paper | ID_Bargmann Database:CX18040 | See Figure 2, Figure 2—figure supplement 1, Figure 4—figure supplement 2 |
| Strain, strain background (C. elegans N2, hermaphrodite) | unc-9 unc-7; AWA,AIA::unc-9(+); AIA::GCaMP5A | this paper | ID_Bargmann Database:CX18041 | See Figure 2, Figure 2—figure supplement 1, Figure 4—figure supplement 2 |
| Strain, strain background (C. elegans N2, hermaphrodite) | unc-7 unc-9; AWA::Chrimson; AIA::GCaMP5A | this paper | ID_BargmannDatabase:CX17320 | See Figure 2, Figure 5—figure supplement 3 |
| Strain, strain background (C. elegans N2, hermaphrodite) | AIA::Chrimson; AIA::GCaMP5A | this paper | ID_BargmannDatabase:CX17432 | See Figure 2, Figure 2—figure supplement 1 |
| Strain, strain background (C. elegans N2, hermaphrodite) | unc-7 unc-9; AIA::Chrimson; AIA::GCaMP5A | this paper | ID_BargmannDatabase:CX17895 | See Figure 2 |
| Strain, strain background (C. elegans N2, hermaphrodite) | AIA::Chrimson; AWA::GCaMP2.2b | this paper | ID_BargmannDatabase:CX17464 | See Figure 2, Figure 2—figure supplement 1 |
| Strain, strain background (C. elegans N2, hermaphrodite) | unc-7 unc-9; AIA::Chrimson; AWA::GCaMP2.2b | this paper | ID_BargmannDatabase:CX17897 | See Figure 2 |
| Strain, strain background (C. elegans N2, hermaphrodite) | unc-18; AIA::Chrimson; AIA::GCaMP5A | this paper | ID_BargmannDatabase:CX17584 | See Figure 2—figure supplement 1 |
| Strain, strain background (C. elegans N2, hermaphrodite) | unc-18; AIA::Chrimson; AWA::GCaMP2.2b | this paper | ID_BargmannDatabase:CX17640 | See Figure 2—figure supplement 1 |
| Strain, strain background (C. elegans N2, hermaphrodite) | unc-13; AIA::GCaMP5A | this paper | ID_BargmannDatabase:CX16591 | See Figure 3, Figure 3—figure supplement 1, Figure 4—figure supplement 2 |
| Strain, strain background (C. elegans N2, hermaphrodite) | unc-18(e234); AIA::GCaMP5A | this paper | ID_BargmannDatabase:CX16412 | See Figure 3, Figure 3—figure supplement 1, Figure 4—figure supplement 2 |
| Strain, strain background (C. elegans N2, hermaphrodite) | unc-13; AWA::Chrimson; AIA::GCaMP5A | this paper | ID_BargmannDatabase:CX16592 | See Figure 3, Figure 4—figure supplement 2 |
| Strain, strain background (C. elegans N2, hermaphrodite) | unc-18(e234); AWA::Chrimson; AIA::GCaMP5A | this paper | ID_BargmannDatabase:CX17158 | See Figure 3, Figure 3—figure supplement 1, Figure 4, Figure 4—figure supplement 2 |
| Strain, strain background (C. elegans N2, hermaphrodite) | unc-18(e81); AWA::Chrimson; AIA::GCaMP5A | this paper | ID_BargmannDatabase:CX17640 | See Figure 3, Figure 3—figure supplement 1, Figure 4, Figure 4—figure supplement 2 |
| Strain, strain background (C. elegans N2, hermaphrodite) | unc-13; AWA::Chrimson; AWA::GCaMP2.2b | this paper | ID_BargmannDatabase:CX17213 | See Figure 3, Figure 3—figure supplement 1 |
| Strain, strain background (C. elegans N2, hermaphrodite) | unc-31; AWA::Chrimson; AIA::GCaMP5A | this paper | ID_BargmannDatabase:CX17319 | See Figure 3—figure supplement 1 |
| Strain, strain background (C. elegans N2, hermaphrodite) | eat-4-FRT; AWA::Chrimson; AIA::GCaMP5A | this paper | ID_BargmannDatabase:CX17714 | See Figure 4, Figure 4—figure supplement 1, Figure 4—figure supplement 2 |
| Strain, strain background (C. elegans N2, hermaphrodite) | eat-4-FRT; AWC,ASE,ASK,ASG::nFlippase; AWA::Chrimson; AIA::GCaMP5A | this paper | ID_BargmannDatabase:CX17679 | See Figure 4, Figure 4—figure supplement 1, Figure 4—figure supplement 2 |
| Strain, strain background (C. elegans N2, hermaphrodite) | eat-4-FRT; ASK::nFlippase; AWA::Chrimson; AIA::GCaMP5A | this paper | ID_BargmannDatabase:CX17722 | See Figure 4, Figure 4—figure supplement 1, Figure 4—figure supplement 2 |
| Strain, strain background (C. elegans N2, hermaphrodite) | eat-4-FRT; ASG::nFlippase; AWA::Chrimson; AIA::GCaMP5A | this paper | ID_BargmannDatabase:CX17892 | See Figure 4, Figure 4—figure supplement 1, Figure 4—figure supplement 2 |
| Strain, strain background (C. elegans N2, hermaphrodite) | eat-4-FRT; AWC::nFlippase; AWA::Chrimson; AIA::GCaMP5A | this paper | ID_BargmannDatabase:CX17611 | See Figure 4, Figure 4—figure supplement 1, Figure 4—figure supplement 2 |
| Strain, strain background (C. elegans N2, hermaphrodite) | eat-4-FRT; AWC,ASE::nFlippase; AWA::Chrimson; AIA::GCaMP5A | this paper | ID_BargmannDatabase:CX17723 | See Figure 4, Figure 4—figure supplement 1, Figure 4—figure supplement 2 |
| Strain, strain background (C. elegans N2, hermaphrodite) | che-1; AWA::Chrimson; AIA::GCaMP5A | this paper | ID_BargmannDatabase:CX17678 | See Figure 4, Figure 4—figure supplement 1 |
| Strain, strain background (C. elegans N2, hermaphrodite) | AWC,ASE,ASK,ASG::nFlippase; AWA::Chrimson; AIA::GCaMP5A | this paper | ID_BargmannDatabase:CX17866 | See Figure 4, Figure 4—figure supplement 1, Figure 4—figure supplement 2 |
| Strain, strain background (C. elegans N2, hermaphrodite) | unc-18; AWC,ASE::unc-18(+); AWA::Chrimson; AIA::GCaMP5A (line A) | this paper | ID_BargmannDatabase:CX17675 | See Figure 4, Figure 4—figure supplement 1 |
| Strain, strain background (C. elegans N2, hermaphrodite) | unc-18; AWC,ASE::unc-18(+); AWA::Chrimson; AIA::GCaMP5A (line B) | this paper | ID_BargmannDatabase:CX17676 | See Figure 4, Figure 4—figure supplement 1 |
| Strain, strain background (C. elegans N2, hermaphrodite) | unc-18; AWC,ASE::unc-18(-); AWA::Chrimson; AIA::GCaMP5A | this paper | ID_BargmannDatabase:CX17677 | See Figure 4, Figure 4—figure supplement 1 |
| Strain, strain background (C. elegans N2, hermaphrodite) | ASK::GCaMP5A | DOI: 10.1016/j.neuron.2019.01.053 | ID_BargmannDatabase:CX17590 | See Figure 5, Figure 5—figure supplement 1, Figure 5—figure supplement 3 |
| Strain, strain background (C. elegans N2, hermaphrodite) | unc-18; ASK::GCaMP5A | this paper | ID_BargmannDatabase:CX17724 | See Figure 5—figure supplement 1 |
| Strain, strain background (C. elegans N2, hermaphrodite) | odr-10; ASK::GCaMP5A | this paper | ID_BargmannDatabase:CX17867 | See Figure 5—figure supplement 1 |
| Strain, strain background (C. elegans N2, hermaphrodite) | AWC::GCaMP5A | this paper | ID_BargmannDatabase:CX17520 | See Figure 5, Figure 5—figure supplement 1, Figure 5—figure supplement 3 |
| Strain, strain background (C. elegans N2, hermaphrodite) | unc-18; AWC::GCaMP5A | this paper | ID_BargmannDatabase:CX17636 | See Figure 5—figure supplement 1 |
| Strain, strain background (C. elegans N2, hermaphrodite) | odr-10; AWC::GCaMP5A | this paper | ID_BargmannDatabase:CX17606 | See Figure 5—figure supplement 1 |
| Strain, strain background (C. elegans N2, hermaphrodite) | ASE::GCaMP3 | this paper | ID_BargmannDatabase:CX14571 | See Figure 5, Figure 5—figure supplement 1 |
| Strain, strain background (C. elegans N2, hermaphrodite) | unc-18; ASE::GCaMP3 | this paper | ID_BargmannDatabase:CX17638 | See Figure 5, Figure 5—figure supplement 1 |
| Strain, strain background (C. elegans N2, hermaphrodite) | odr-10; ASE::GCaMP3 | this paper | ID_BargmannDatabase:CX16497 | See Figure 5, Figure 5—figure supplement 1 |
| Strain, strain background (C. elegans N2, hermaphrodite) | ASH::GCaMP3 | DOI: 10.1016/j.neuron.2013.11.020 | ID_BargmannDatabase:CX10979 | See Figure 5—figure supplement 1 |
| Strain, strain background (C. elegans N2, hermaphrodite) | AWA::Chrimson; ASK::GCaMP5A | this paper | ID_BargmannDatabase:CX17751 | See Figure 5—figure supplement 1 |
| Strain, strain background (C. elegans N2, hermaphrodite) | AWA::Chrimson; AWC::GCaMP5A | this paper | ID_BargmannDatabase:CX17521 | See Figure 5—figure supplement 1 |
| Strain, strain background (C. elegans N2, hermaphrodite) | AWA::Chrimson; ASE::GCaMP3 | this paper | ID_BargmannDatabase:CX17392 | See Figure 5—figure supplement 1 |
| Strain, strain background (C. elegans N2, hermaphrodite) | ceh-36; AIA::GCaMP5A | DOI: 10.1016/j.celrep.2015.08.032 | ID_BargmannDatabase:CX16169 | See Figure 5, Figure 5—figure supplement 3 |
| Strain, strain background (C. elegans N2, hermaphrodite) | odr-7 ceh-36; AIA::GCaMP5A | DOI:10.1016/j.celrep.2015.08.032 | See Figure 5, Figure 5—figure supplement 3 | |
| Strain, strain background (C. elegans N2, hermaphrodite) | ASK,AWA::GCaMP6s; AIA::GCaMP5A | this paper | ID_BargmannDatabase:CX18038 | See Figure 5—figure supplement 2 |
| Strain, strain background (C. elegans N2, hermaphrodite) | AIA::GFP | this paper | ID_BargmannDatabase:CX8293 | See Figure 6, Figure 6—figure supplement 1 |
| Strain, strain background (C. elegans N2, hermaphrodite) | AIA::GCaMP5A | this paper | ID_BargmannDatabase:CX16976 | See Figure 6, Figure 6—figure supplement 1 |
| Chemical compound, drug | (-)-tetramisole hydrochloride | Sigma | L9756 | CAS 16595-80-5 |
| Chemical compound, drug | Polydimethylsiloxane (PDMS) | Sigma | 761036 | 9:1 base:curing agent, Sylgard 184 |
| Software, algorithm | ImageJ | ImageJ (http://imagej.nih.gov/ij/) | RRID:SCR_003070 | Version 1.52a |
| Software, algorithm | GraphPad Prism | GraphPad Prism (https://graphpad.com) | RRID:SCR_002798 | Version 8 |
| Software, algorithm | Matlab | MathWorks (https://www.mathworks.com/) | RRID:SCR_001622 | Versions R2013b and R2015a |
| Software, algorithm | Metamorph | Molecular Devices (https://www.moleculardevices.com) | RRID:SCR_002368 | Versions 7.7.6 and 7.7.8 |
| Software, algrorithm | analysis code | this paper | See Source code 1 |
Additional files
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Source code 1
Source code for data analysis.
- https://cdn.elifesciences.org/articles/50566/elife-50566-code1-v2.m
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Supplementary file 1
Presynaptic partners of AIA.
Both Chen et al. (2006) and Cook et al. (2019) are based on same collection of serial-section electron micrographs from White et al. (1986). Neurotransmitter information is based on Pereira et al. (2015).
- https://cdn.elifesciences.org/articles/50566/elife-50566-supp1-v2.docx
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Supplementary file 2
Cumulative response time profiles.
Kolmogorov-Smirnov test statistics and sample sizes for all cumulative response time profiles presented, calculated for full 10 s stimulus pulse. Italics indicate non-WT genetic backgrounds. D test represents the maximum effect size across the distributions. p-values below 0.05 are bolded for emphasis.
- https://cdn.elifesciences.org/articles/50566/elife-50566-supp2-v2.docx
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Supplementary file 3
Calcium response magnitude comparisons.
Magnitudes of responses to various stimuli, with either an unpaired t-test (if the number of comparisons is one) or an ordinary one-way ANOVA with Dunnett’s multiple comparisons test (if the number of comparisons exceeds one); * indicates paired t-test. Bolded genotype or stimulus indicates the control group used for comparisons. Italics indicate non-wildtype genetic background. p-values below 0.05 are bolded for emphasis.
- https://cdn.elifesciences.org/articles/50566/elife-50566-supp3-v2.docx
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Supplementary file 4
Calcium rise time comparisons.
Rise times (t66-t33) of responses to various stimuli, with either an ordinary one-way ANOVA with Dunnett’s multiple comparisons test. Bolded genotype or stimulus indicates the control group used for comparisons. Italics indicate non-wildtype genetic background.
- https://cdn.elifesciences.org/articles/50566/elife-50566-supp4-v2.docx
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Supplementary file 5
Strain list.
- https://cdn.elifesciences.org/articles/50566/elife-50566-supp5-v2.docx
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Transparent reporting form
- https://cdn.elifesciences.org/articles/50566/elife-50566-transrepform-v2.docx
