A neural command circuit for grooming movement control
- Howard Hughes Medical Institute, United States
- University of California, Santa Barbara, United States
Figures
Figure 1 with 4 supplements
Sensory neurons that elicit antennal grooming.
(A) Grooming movements performed by flies in which aJO spGAL4 pairs drove expression of thermally activated dTrpA1. Movements were manually scored from 2 min of recorded video per fly (n ≥ 17 flies …
Figure 1—figure supplement 1
GAL4 lines that target expression to sensory neurons from the antennae and elicit grooming.
(A) Percent of total time three GAL4 lines expressing dTrpA1 spent antennal grooming, compared with controls. Box plots, statistics, and experimental conditions are as described in Figure 1A,B (n ≥ …
Figure 1—figure supplement 2
spGAL4 pairs that target expression to sensory neurons in the antennae and elicit grooming.
(A–E) CNS expression patterns of spGAL4 line pairs that displayed increased antennal grooming with dTrpA1. (A′–E′) Native GFP expression (green) in the antennae of corresponding spGAL4 lines shown …
Figure 1—figure supplement 3
JO neurons projecting to zone C/E elicit antennal grooming.
(A, B) Co-expression of JO sensory neurons using aJO-LexA to express GFP (green) and (A) JO4-GAL4 or (B) JO31-GAL4 lines to express tdTomato (magenta). See Figure 4—figure supplement 1 for more …
Figure 1—figure supplement 4
Most stochastically labeled aJO neurons show projections to both the AMMC and ventral SEZ.
(A) Overview of the aJO neuron population with AMMC, posterior, and ventral SEZ projections (white, yellow, and red arrows point to each projection respectively). (B–F) Multicolor stochastic …
Figure 2 with 2 supplements
Interneurons that elicit antennal grooming.
(A) Grooming movements performed by interneuron spGAL4 pairs expressing thermally activated dTrpA1. Data was obtained and displayed as described in Figure 1A. (B) Percent time flies spent antennal …
Figure 2—figure supplement 1
GAL4 lines that elicit antennal grooming.
(A) Grooming movements performed by GAL4 lines expressing thermally activated dTrpA1. Data was obtained and displayed as described in Figure 1A (n = 10 per line). Note: R11B11-GAL4 did not perform …
Figure 2—figure supplement 2
spGAL4 lines with interneuron expression that elicit antennal grooming.
(A–I) spGAL4 lines expressing GFP (green) are co-stained with anti-GFP (green) with anti-bruchpilot (magenta). The activation domains (ADs) and DNA binding domains (DBDs) used are shown below each …
Figure 3
Neurons that elicit antennal grooming have neurites in the AMMC and/or SEZ.
(A–D) spGAL4 pairs targeting each neuronal class in the ventral brain: (A) aJO-spGAL4-1, (B) aBN1-spGAL4-1, (C) aBN2-spGAL4-2, and (D) aDN1-spGAL4-1. aDN1 is shown as an example in (D), but there …
Figure 4 with 3 supplements
Functional relationships among putative antennal circuit components.
(A) Overview of experiments shown in (B, E). Grooming was induced by thermogenetic activation of Johnston's Organ (JO) neurons (dTrpA1) or by imposed displacements of the antennae. Synaptic release …
Figure 4—figure supplement 1
aJO-, aBN2-, and aDN-LexA lines.
(A) Percent of total time that LexA lines expressing thermally activated dTrpA1 spent antennal grooming. Box plots, statistics, and experimental conditions are as described in Figure 1A,B (n > 10; …
Figure 4—figure supplement 2
Co-expression of LexA lines with selected spGAL4 pairs.
(A–F) Co-expression of LexA and spGAL4 lines. Left column: spGAL4 expression patterns. Middle column: LexA expression patterns. Right column: Merged expression patterns. (A, B) Co-expression of …
Figure 4—figure supplement 3
Testing of stimulus parameters for the antennal displacement assay.
(A–C) Flies were prepared as shown in Figure 4C,D (see ‘Materials and methods’) and tested for their grooming responses when different magnetic field frequencies (A) or voltages (B) were applied to …
Figure 5 with 2 supplements
Antennal grooming neurons are in close proximity.
(A–H) Co-expression in neuronal pairs using two binary expression systems (LexA and spGAL4) to express tdTomato or GFP in each neuronal class. Processed maximum intensity projections of frontal and …
Figure 5—figure supplement 1
GFP-positive staining indicates reconstitution across synaptic partner (GRASP) staining indicates close proximity of neurons involved in antennal grooming.
(A–H) Expression of different GFP halves (spGFP11 and spGFP1-10) in putative connected neuronal pairs. aJO-LexA drove expression of spGFP11 in the JO neurons and spGFP1-10 expression was driven by …
Figure 5—figure supplement 2
Co-staining indicates close proximity of neurons involved in antennal grooming.
(A–H) Unprocessed maximum intensity projections of co-stained images shown in Figure 5 (see ‘Materials and methods’).
Figure 6 with 3 supplements
Different antennal grooming neurons are functionally connected.
(A–I) Dissected CNSs with different neuronal classes expressing CsChrimson (magenta) were activated with red light while changes in calcium in their putative downstream partners expressing GCaMP6 …
Figure 6—figure supplement 1
Functional connectivity: controls, technical details, and raw data.
(A) Control experiments for CsChrimson/GCaMP6s activation. LexAop-CsChrimson was crossed with the control LexA driver, and GCaMP6s was expressed with aBN2-spGAL4-1 (top) or aDN2-spGAL4-2 (bottom). …
Figure 6—figure supplement 2
Raw data for functional connectivity experiments (at low intensity red light).
Raw data for experiments shown in Figure 6. All experiments shown were done in the absence of drugs. Red light intensity was set at 50 μW/mm2. Each column corresponds to the number of light pulses …
Figure 6—figure supplement 3
Raw data for functional connectivity experiments (at high intensity red light).
Raw data for experiments shown in Figure 6. All experiments shown were done in the absence of drugs. Red light intensity was set betwen 290 to 700 μW/mm2. Each column corresponds to the number of …
Figure 7 with 1 supplement
A circuit whose components elicit different antennal grooming durations.
(A) The antennal grooming circuit (lateral view of tracings). Specific colors represent each neuron type shown in B. (B) Wiring diagram of the circuit. Lightning bolt represents mechanical …
Figure 7—figure supplement 1
CsChrimson activation of different neuronal classes.
(A) Stacked ethograms of grooming movements performed by the spGAL4 pairs indicated expressing CsChrimson (10–15 flies shown for each spGAL4 pair). Gray bars indicate when the red light was on. …
Videos
Video 1
Traced antennal grooming circuit.
Traced and manually aligned neurons are shown in different colors. aJO neurons (blue) project from the second antennal segment into the anterior brain. aBN1 (red) has cell bodies in the dorsal and …
Video 2
Computationally aligned antennal grooming circuit.
Computationally aligned neurons are shown in different colors. aJO neurons (blue), aBN1 (red), aBN2 (yellow) and aDN1 (green). The neuropil was stained with anti-bruchpilot (grey). See ‘Materials …
Video 3
Grooming movements performed in response to displacements of the antennae.
The third antennal segments of a control fly were coated with iron powder, and the fly was tethered within the electromagnetic apparatus shown in Figure 4C,D. The infrared light positioned behind …
Video 4
Grooming in response to red light stimulation of CsChrimson-expressing aBN1 neurons.
CsChrimson was expressed in aBN1 using aBN1-spGAL4-1. The infrared light in the bottom right hand corner shows when the red light was on to activate aBN1. Note that grooming persists upon cessation …
Video 5
Grooming in response to red light stimulation of CsChrimson-expressing aDN2 neurons.
CsChrimson was expressed in aDN2 using aDN2-spGAL4-2. The infrared light in the bottom right hand corner shows when the red light was on to activate aDN2. Note that grooming does not persist upon …
Video 6
Grooming in response to red light stimulation of CsChrimson-expressing aJO neurons.
CsChrimson was expressed in the aJO using aJO-spGAL4-1. The infrared light in the bottom right hand corner shows when the red light was on to activate the aJO. Note that grooming persists upon …
Additional files
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Supplementary file 1
Enhancer identities used to target different neuronal classes.
- https://doi.org/10.7554/eLife.08758.031
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Supplementary file 2
Stimulus conditions used for the experiment shown in Figure 6.
- https://doi.org/10.7554/eLife.08758.032
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Supplementary file 3
Stocks used in this study.
- https://doi.org/10.7554/eLife.08758.033
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Supplementary file 4
Design plans for the fly electromagnetic stimulation rig.
- https://doi.org/10.7554/eLife.08758.034
