Global change in brain state during spontaneous and forced walk in Drosophila is composed of combined activity patterns of different neuron classes
- School of Life Sciences, Technical University of Munich, Germany
- University of Bonn, Medical Faculty (UKB), Institute of Physiology II, Germany
- Max Planck Institute for Brain Research, Computation in Neural Circuits, Germany
Figures
Figure 1 with 1 supplement
Global brain activation during walk.
(A) Schematic overview of the preparation and analysis method. Please see methods for details. (B) Raster plot of the activity of regions. Top panels depict walking bouts in green and rest or …
Figure 1—figure supplement 1
Correlation of regional activity with walk.
(A) R2 during walk at different recording frequencies for pan-neuronally expressed GCaMP6f or GCaMP6m (all regions were pooled). No significant difference was found for walk between the different …
Figure 2 with 1 supplement
Activity of neurons releasing the three major neurotransmitters, glutamate, GABA, and acetylcholine, during walk.
N = 5 flies for each genotype. (A) R2 for regression of single region activity with walk for different genotypes (all regions were pooled). No pairwise comparison of fly-wise averages is …
Figure 2—figure supplement 1
Correlation coefficients for regression of single region activity with walk.
N = 5 flies for each genotype. Empty markers correspond to adjusted (Benjamini–Hochberg correction) p-value >0.05 for comparison to 0. Right, expression pattern for three example flies per genotype …
Figure 3 with 1 supplement
Neuromodulatory neurons are strongly and differentially activated during walk.
TH/DDC-Gal4: N = 9, Tdc2-Gal4: N = 7, Trh-Gal4: N = 6 flies. (A) R2 for regression of single region activity with walk for different genotypes (all regions were pooled). Mann–Whitney U-test …
Figure 3—figure supplement 1
Correlation coefficients for regression of single region activity with walk.
Empty markers correspond to adjusted (Benjamini–Hochberg correction) p-value >0.05 (null hypothesis is 0). TH/DDC-Gal4: N = 9, Tdc2-Gal4: N = 7. (A) Dopaminergic neurons (TH/DDC-Gal4 or …
Figure 4 with 1 supplement
Whole brain analysis pinpoints specific subregions responding to walk.
(A) Images of example components that are significantly correlated with walk (coefficient or R2 95% CI above zero). Upper left: Components derived from imaging with a pan-neuronal driver. Upper …
Figure 4—figure supplement 1
Component maps and correlation with walk.
(A) Z-stacks of functional components for walk mapped onto the adult fly brain. (B) Functional components for walk and their names or acronyms. See Table 2 for definition of acronyms. (C) R2 for …
Figure 5 with 1 supplement
Turning activates specific components and candidate neurons.
(A) Examples of components present in both the left and right hemispheres labeled in different colors (magenta and green). Panels on the right present an example component that could be mapped to a …
Figure 5—figure supplement 1
Turning activates specific components and candidate neurons.
(A) Examples of components present in both the left and right hemispheres labeled in different colors (magenta and green). (B) R2 of regression coefficients during turn for all genotypes. The value …
Figure 6 with 2 supplements
Onset dynamics of activity in multiple functional components across the brain at start of walk.
Walk-onset triggered average activity for pan-neuronal data in individual active components at 30 Hz or higher temporal resolution. Trials were normalized individually before averaging (see Figure …
Figure 6—figure supplement 1
Graphs show onset of walk trials for all flies and all transgenic lines at all recording speeds.
The gray band represents the uncertainty of walk-onset time due to lower temporal resolution in some recordings. A corrective factor based on regression coefficient with walk (fluorescence response …
Figure 6—figure supplement 2
Normalized trials ∆F/F GCaMP fluorescence in brain regions 1 s before to 1 s after walk onset for different anatomically defined brain regions: (A) dopaminergic neurons (TH/DDC-Gal4;UAS-GCaMP), (B) octopaminergic neurons (Tdc2-Gal4;UAS-GCaMP), and (C) serotonergic neurons (Trh-Gal4;UAS-GCaMP).
Gray shading corresponds to the standard deviation. Red band is the onset of walk.
Figure 7 with 1 supplement
Forced and spontaneous walk elicit highly similar global brain activity.
(A) Z-stacks of map of brain regions activated by walk in flies expressing cytosolic GCaMP (pan-Gal4;UAS-GCaMP6m) or synaptically tagged GCaMP (pan-Gal4;UAS-sytGCaMP6m) (R2 of ). (B) Time series …
Figure 7—figure supplement 1
Additional evidence for most activity being induced by walk.
(A) Comparison of activity during walk for cytosolic and synaptically tagged GCaMP. Left: expression patterns (baseline fluorescence), Right: Coefficient in linear models of regional activity as a …
Videos
Video 1
Movie of pan-neuronal activation during walk and groom (accelerated).
Video 2
Movie of TH/DDC-neuronal activation during walk (accelerated).
Video 3
Movie of TDC2-neuronal activation during walk (accelerated).
Video 4
Movie of Trh-neuronal activation during walk (accelerated).
Video 5
Video sequence showing activation of the PPM2-LW neurons during turning.
Video 6
Video showing neuronal activation moving from ventral to dorsal brain areas.
Note that a clear progression of activation was observed only in a fraction of trials.
Video 7
Video showing whole brain activity of a fly being forced to walk on a rotating rod.
Video 8
Video showing whole brain activity of a fly with a severed connection between the brain and ventral nerve cord (VNC) being forced to walk on a treadmill.
Note that the fly is still capable of walking on the rod when being forced. We did not, however, observe spontaneous walking activity.
Tables
Table 1
Summary statistics of R2 dependence on different factors where R2 is obtained from regressing a single regionally averaged brain activity with a single behavior.
| sum_sq | PR(>F) | |
|---|---|---|
| GAL4 | 0.002197 | 7.58E−01 |
| Behavior | 12.215185 | 1.64E−106 |
| RegionNames | 10.372902 | 4.13E−65 |
| UAS | 2.083138 | 2.13E−18 |
| Behavior:RegionNames | 1.45213 | 9.17E−01 |
| Residual | 67.904784 | NaN |
Table 2
List of components with potential underlying candidate neurons.
Most matches are speculative except those in bold that correspond to high confidence matches due to the lack of other Gal4-positive neurons in the region.
| Short name | Description | Examples of matching (Flycircuit) neurons | Present in Gal4 line (detected in ≥5 flies) |
|---|---|---|---|
| Antennal lobe and mushroom body | |||
| AL | Whole antennal lobe | Gad1-F-000394; Gad1-F-100601; Gad1-F-800129; VGlut-F-900126 | Nsyb, GMR57C10, Vglut, Gad |
| PN | Antennal lobe projection neuron | VGlut-F-500486 | Nsyb, GMR57C10, Cha, Vglut, Gad |
| MultiGl | Whole antennal lobe and PN-like projections | Trh-F-600017 | TH, Trh, TDC |
| PNv | Antennal lobe projection neuron, ventral tract of the lateral horn | VFB_00101133 | Nsyb, GMR57C10, Cha, Vglut, Gad |
| PN-KC | Antennal lobe projection neuron and kenyon cell in the same component | Nsyb, GMR57C10, Cha, Vglut | |
| KCab | Alpha-Beta Kenyon cell | Cha-F-300226; Cha-F-100049; fru-F-000026; Vglut-F-100284; Gad1-F-100014; | Nsyb, GMR57C10 |
| KCapbp | Alpha’-Beta’; Kenyon cell | Gad1-F-100024; Trh-F-200069; | Nsyb, GMR57C10 |
| KCg | Gamma kenyon cells | fru-F-000006; Vglut-F-100359; Gad1-F-100021; | Nsyb, GMR57C10 |
| Beta1Betap1 | Beta1 and/or Beta’1 mushroom body compartment | PAM10(B1)_L (FlyEM-HB:1328522741) [VFB_jrchk385] | TH |
| Beta2Betap2 | Beta2 and/or Beta’2 mushroom body compartment | PAM02(B’2a)_L (FlyEM-HB:1295566429) | Nsyb, Gad, TH |
| Gamma1 | Gamma1 mushroom body compartment | PPL1-gamma1-pedc | TH |
| Gamma2 | Gamma2 mushroom body compartment | PPL1-gamma2-alpha’1 | TH |
| Gamma3 | Gamma3 mushroom body compartment | MBON-γ3, PAM-γ3 | Gad, TH |
| Gamma4 | Gamma4 mushroom body compartment | PAM-γ4 | TH |
| Gamma5 | Gamma5 mushroom body compartment | PAM-γ5 | TH |
| SLP-Alpha | Alpha or Alpha’ lobe with projection through the superior lateral neuropil | VGlut-F-500002 | Vglut |
| Alpha1 | Alpha1 mushroom body compartment | PAM-alpha1 | TH |
| Alpha2 | Alpha2 mushroom body compartment | PPL1-alpha’2alpha2 | TH |
| Alpha3 | Alpha3 mushroom body compartment | PPL1-alpha3 | TH |
| Alphap3 | Alpha’3 mushroom body compartment | PPL1-alpha’3 | TH |
| Superior neuropil | |||
| SCLtract | Tract linking both superior clamp | Trh-F-200082,Trh-F-100051 | Trh |
| CLvert | Lateral part of the superior clamp | TH-F-000023 | Nsyb, TH |
| CL? | Interrogation point surrounding the pedonculus, | Trh-F-300074,Trh-M-700081, DNp32 | TH, Trh |
| CL-LH | Surrounds the lateral horn from the medial and ventral directions | Trh-F-200047 | Trh, TDC |
| CL | Other shapes at the level of the clamp | Gad1-F-700550,Gad1-F-800055 | Nsyb, GMR57C10, Gad, TH, TDC |
| SMPm | Medial part of the superior medial protocerebrum | TH-F-000021; VGlut-F-700286, Cha-F-300251 | Nsyb, GMR57C10, Cha, Vglut, Gad, TH, TDC |
| PPL-SMP | Ventral lateral part of the superior medial protocerebrum, with tracts coming from a posterior lateral cell cluster | TH-F-000019,TH-F-000018,TH-F-000046 | TH |
| SMPl-SIP | Superior intermediate protocerebrum and lateral part of the superior medial protocerebrum | Cha-F-000221,Cha-F-300154, TH-F-300056 | Nsyb, GMR57C10, Cha, Gad, TH, TDC |
| SIP-SMPd | Superior intermediate protocerebrum and dorsal part of the superior medial protocerebrum | fru-F-800063 | Nsyb |
| SIP-FB | Superior intermediate protocerebrum and dorsal layer of the fan-shaped body | Trh-F-100015 | Nsyb, GMR57C10, TH, Trh |
| FB-SN | Broad innervaion of the superior neuropil, and fan-shaped body | OA-VMP3, OA-VPM4 | TDC |
| SLP-SMPproj | Large SMP neuron projecting to ventral regions | Trh-F-700011,Trh-F-000083 | Nsyb, GMR57C10, TH, Trh |
| SLP-SMP | Superior lateral protocerebrum and superior medial protocerebrum | Trh-F-500176, DNp25 | Trh |
| SLP | Superior lateral protocerebrum only | TH-F-100046 | Nsyb, GMR57C10, TH, TDC |
| LH-SLP | Lateral horn and superior lateral protocerebrum | Gad1-F-900346,Gad1-F-600340 | Nsyb, Gad |
| Central complex | |||
| FBcol | Fan-shaped body columns | Tdc2-F-100009; Tdc2-F-300026; Tdc2-F-300001; Tdc2-F-200011; Tdc2-F-100062; Tdc2-F-100016; Gad1-F-900245; Gad1-F-800329; Gad1-F-500513; Gad1-100157 | Nsyb, GMR57C10, Vglut, Gad, TDC |
| FBlayv | Ventral layer of the fan-shaped body | TH-M-300065 | Nsyb, TH |
| FBlaym | Medial layer of the fan-shaped body | TH-F-200055 | Nsyb, TH |
| FBlayd | Dorsal layer of the fan-shaped body | TH-F-200054; Trh-F-300036; Trh-F-400062 | Nsyb, TH, Trh |
| NO | Nodulus or noduli | Cha-F-100429 | Nsyb |
| PB | Protocerebral bridge only | Vglut-F-800282; Vglut-F-600784; Vglut-F-600229; Gad1-F-600267; Gad1-F-100361; Gad1-F-100593; Vglut-F-000156; Vglut-F-100064 | Nsyb, Vglut, Gad |
| PBfull | Components with full protocerebral bridge | Cha-F-900016, Cha-F-200148 | Nsyb, GMR57C10, Cha |
| PB-DA | Protocerebral bridge and two dots (maybe cell bodies) at the top of the trachea | TH-F-000048 | TH |
| BU-PBl-EB | Bulb, ellipsoid body and lateral part of the protocerebral bridge | Gad1-F-900445 | Nsyb, Gad |
| PB-EB | EB-radial and PB glomeruli | Cha-F-500009 | Nsyb, GMR57C10, Cha, Gad |
| EB | Ellipsoid body rings | Trh-F-300095; Cha-F-800146 | Nsyb, GMR57C10, Cha, Vglut, Trh |
| EB-DA | Ellipsoid body and lateral accessory lobe | TH-F-100001 | TH |
| AOTU-BU | Anterior optic tubercule and bulb | Gad1-F-200712, VGlut-F-400630 | Nsyb, GMR57C10, Cha, Vglut, Gad |
| Posterior neuropil | |||
| IB | Inferior bridge | Nsyb, Gad | |
| ATL | Antler | Adult antler neuron 031 | TH, Trh |
| M-Omega | Posterior ensemble forming an M dorsally and an omega ventrally | TH-F-300078 | TH |
| SPS | Superior posterior slope | VGlut-F-900089, VGlut-F-800136,Cha-F-800003,Gad1-F-900039 | Nsyb, GMR57C10, Cha, Vglut, Gad |
| IPS-Y | Inverse Y shape in the posterior slope | DNb02? | Nsyb, GMR57C10, Cha, Vglut, Gad |
| LAL-PS | Lateral accessory lobe and posterior slope | DNb01? | Nsyb, GMR57C10, Cha, Vglut, Gad |
| PPM2-LW | PPM2-LAL-We L,R | TH-F-000000,TH-F-000015,TH-F-000016 | TH |
| PPM2-VI | PPM2-VMNP-INP L,R | TH-F-000007,TH-F-300058 | TH |
| Lateral neuropil | |||
| WPENb | Antennal mechanosensory and motor center and/or Wedge, in the posterior lateral protocerebrum and posterior connection to opposite side | VGlut-F-200005,WPNb, WPNB3#5 (FAFB:4271367) [VFB_001011lp] | Vglut, Gad |
| AMMC-PLP | Antennal mechanosensory and motor center and/or Wedge and branch in the posterior lateral protocerebrum | VGlut-F-400269 | Nsyb, Cha, Vglut, Gad |
| AMMC-WE | Antennal mechanosensory and motor center and/or Wedge | VGlut-F-000138, VGlut-F-400586 | Nsyb, GMR57C10, Vglut, Gad |
| WE-DA | Wedge with two branches forming a large V | TH-F-200127,TH-F-000024 | TH |
| AVLPonlyproj | Lowest medial part of the anterior ventral lateral protocerebrum projecting ventrally | Cha-F-700097 | |
| AVLPprojm | Lowest medial part of the anterior ventral lateral protocerebrum projecting ventrally | Gad1-F-500762, Cha-F-400059; Gad1-F-000013 | Nsyb, GMR57C10, Gad |
| AVLPprojl | Lowest lateral part of the anterior ventral lateral protocerebrum projecting ventrally | Cha-F-800125 | Cha |
| AVLPm | Anterior ventral lateral protocerebrum medial part | Gad1-F-900529, Cha-F-800062,Trh-F-400039,Trh-F-400070; Vglut-F-200405*; Vglut-F-900122; Cha-F-400237; Cha-F-200299; Gad1-F-500279 | Nsyb, GMR57C10, Vglut, Gad, TH, Trh, TDC |
| AVLPd | Anterior ventral lateral protocerebrum dorsal part | Cha-F-000424 | Nsyb |
| AVLPshell | Anterior ventral lateral protocerebrum surface | Trh-F-100082 | TH, Trh |
| AVLPsmear | Anterior ventral lateral protocerebrum anterior part | Nsyb | |
| VLPl | Ventro-lateral protocerebrum most lateral part | Gad1-F-900096, Vglut-F-500616,Cha-F-800087 | Nsyb, GMR57C10, Cha, Vglut, Gad |
| PLP-LH | Posterior lateral protocerebrum to the basis of the lateral horn | Gad1-F-500325 | Nsyb |
| PLP | Posterior lateral protocerebrum | Gad1-F-800092 | Nsyb, Gad |
| Ventral neuropil | |||
| PI | Pars intercerebralis | Trh-F-100040, Trh-M-000056 | Nsyb, GMR57C10, TH, Trh |
| PI-PRW | Pars intercerebralis connected to Prow | VGlut-F-600158 | Nsyb, GMR57C10, Gad |
| PRW | Prow | TH-M-000037 | Nsyb, GMR57C10, Vglut, Gad, TH |
| PRW-SLP | Prow and superior lateral protocerebrum | Gad1-F-600213, Cha-F-200258; fru-F-000133; Gad1-F-600213; Trh-F-100091 | Nsyb, GMR57C10 |
| PENP-CL | Periesophageal neuropils and clamp | mALD3_L (FlyEM-HB:822708945) | Nsyb, GMR57C10, Cha, Gad |
| GNGvw | Gnathal ganglia medial and lateral | Cha-F-400186 | Cha |
| GNGm | Gnathal ganglia medial | Cha-F-300235 | Nsyb, GMR57C10 |
| GNGml | Gnathal ganglia medial–lateral | Cha-F-400159 | Nsyb, GMR57C10, Cha, Gad |
| GNGl | Gnathal ganglia lateral | Cha-F-400146 | Nsyb, Cha |
| GNG-AMMC | Gnathal ganglia and on the opposite side antennal mechanosensory and motor center and posterior lateral protocebrum | VGlut-F-600685 | Nsyb, Vglut |
| GNGva | Ventral anterior part of the gnathal ganglia | Nsyb | |
| vaCells | Ventral anterior cells | TH-F-100049 | TH, Trh |
| Optic lobe | |||
| OL | Optic lobe; mostly medulla and lobulla | Nsyb, GMR57C10, Cha, Vglut, Gad | |
| LOP | Lobulla plate | Cha-F-600161 | Cha, TDC |
| OL-FB | Optic lobe to central regions including the fan-shaped body | TH | |
| OL-PENP | Optic lobe and periesophageal neuropils | Tdc2-F-200056 | Nsyb, Gad, Trh, TDC |
| OL-PLP | Optic lobe and posterior lateral protocerebrum | Cha-F-000316 | Nsyb, Cha, Gad |
| OL-WE | Optic lobe and wedge | TH-F-300030 | TH |
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Genetic reagent (D. melanogaster) | UAS-GCaMP6s | Bloomington Drosophila Stock Center | BDSC_42749 | |
| Genetic reagent (D. melanogaster) | UAS-GCaMP6m | Bloomington Drosophila Stock Center | BDSC_42750 | |
| Genetic reagent (D. melanogaster) | UAS-GCaMP7s | Bloomington Drosophila Stock Center | BDSC_79032 | |
| Genetic reagent (D. melanogaster) | UAS-GCaMP6f | Bloomington Drosophila Stock Center | BDSC_42747 | |
| Genetic reagent (D. melanogaster) | UAS-GCaMP7f | Bloomington Drosophila Stock Center | BDSC_79031 | |
| Genetic reagent (D. melanogaster) | UAS-syt-GCaMP6s | Vanessa Ruta | N/A | |
| Genetic reagent (D. melanogaster) | GMR58E04-Gal4 | Bloomington Drosophila Stock Center | BDSC_41347 | |
| Genetic reagent (D. melanogaster) | TH-Gal4 | Bloomington Drosophila Stock Center | BDSC_8848 | |
| Genetic reagent (D. melanogaster) | Tdc2-Gal4 | Bloomington Drosophila Stock Center | BDSC_9313 | |
| Genetic reagent (D. melanogaster) | Trh-Gal4 | Bloomington Drosophila Stock Center | BDSC_38388 | |
| Genetic reagent (D. melanogaster) | Ddc-Gal4 | Bloomington Drosophila Stock Center | BDSC_7009 | |
| Genetic reagent (D. melanogaster) | TH, 58E02-Gal4 | Siju et al. | N/A | |
| Genetic reagent (D. melanogaster) | TH, DDC-Gal4 | This paper | N/A | |
| Genetic reagent (D. melanogaster) | Vglut-Gal4 | Bloomington Drosophila Stock Center | BDSC_24635 | |
| Genetic reagent (D. melanogaster) | Cha-Gal4 | Bloomington Drosophila Stock Center | BDSC_6798 | |
| Genetic reagent (D. melanogaster) | Nsyb-Gal4 | Bloomington Drosophila Stock Center | BDSC_51635 | |
| Genetic reagent (D. melanogaster) | GMR57C10-Gal4 | Bloomington Drosophila Stock Center | BDSC_39171 | |
| Genetic reagent (D. melanogaster) | Gad1-Gal4 | Bloomington Drosophila Stock Center | BDSC_51630 | |
| Software, algorithm | Python 3 | Python Software Foundation | https://www.python.org | |
| Software, algorithm | ImageJ/FIJI | Schindelin et al., 2012 | https://fiji.sc/ | |
| Software, algorithm | MATLAB | MATLAB | mathworks.com | |
| Software, algorithm | Analysis code | GitHub | https://github.com/sophie63/Aimon2022 |
Additional files
-
MDAR checklist
- https://cdn.elifesciences.org/articles/85202/elife-85202-mdarchecklist1-v2.docx
-
Supplementary file 1
Description of components and putative underlying neurons.
- https://cdn.elifesciences.org/articles/85202/elife-85202-supp1-v2.docx
