foxQ2 marks fast-acting interneurons including dopaminergic neurons of mushroom bodies and central complex in the beetle Tribolium castaneum
- Department of Evolutionary Developmental Genetics, University of Göttingen, GZMB, Germany
- Abberior Instruments GmbH, Germany
- Institute of Zoology, Experimental Morphology and Neuroanatomy, University of Cologne, Germany
- Institut de Génomique Fonctionnelle de Lyon (IGFL), École Normale Supérieure de Lyon, Centre National de la Recherche Scientifique (CNRS) and Université Lyon 1, France
Figures
Figure 1 with 2 supplements
Overview on the Tc-foxQ2II-positive cells.
Neuronal projection patterns were identified using immunohistochemistry against EGFP in the adult brain of foxQ2-5’-line. (A–D) Different parts of the adult brain stained for EGFP (green), synapsin (magenta), and DAPI (blue). (A’–D’) The EGFP channel is shown with the brain outline indicated by dashed lines. (A–A’) Overview of a projection of the entire brain; (B–B’) anterior subsection; (C–C’) medial subsection; (D–D’) posterior subsection. White arrowhead: lateral cluster 1; white arrow: lateral cluster 2; open arrowhead: lateral cluster 3; open arrow: lateral cluster 4; yellow arrowhead: median cluster 1; yellow arrow: median cluster 2; yellow dashed circle: posterior cluster; white dashed circle: tritocerebrum cluster; white star: anterior cluster. Scale bar: 100 μm.
Figure 1—figure supplement 1
Characterization of the foxQ2-5’-imaging line.
Tc-FoxQ2 showed a high degree of overlap with EGFP in adult brains in co-immunostaining of brain sections using antibodies against FoxQ2 and EGFP. (A) Cells co-expressing FoxQ2 and EGFP are indicated by white arrows. Green represents EGFP, while magenta represents FoxQ2. (B) The Venn diagram illustrates the number of cells expressing only EGFP, only FoxQ2, and those that are double-positive for EGFP and FoxQ2. The proportion of co-expressing cells relative to all EGFP- or FoxQ2-expressing cells was 79.0% for EGFP-positive cells and 85.1% for FoxQ2-positive cells. When removing the completely Tc-FoxQ2-negative AC cluster from the count (white star in Figure 2), 80.0% of the EGFP-positive cells were Tc-FoxQ2 positive, while 85.1% of FoxQ2-expressing cells overlapped with EGFP (n=3). Scale bar: 100 μm.
Figure 1—figure supplement 2
The anterior cluster is not marked by the FoxQ2 antibody and only expresses EGFP.
(A–A’’) Overview of the anterior section of the adult brain stained with EGFP and FoxQ2 antibodies. EGFP is shown in green and FoxQ2 in magenta. (B–B’’) A magnified view of the white-dashed square. The anterior cluster expresses only EGFP. Therefore, we did not consider these cells further. Scale bar: 100 μm.
Figure 2
Overview on the reconstruction of the nine Tc-foxQ2II-positive cell clusters.
(A) 3D reconstruction of major neuropils and EGFP-positive clusters of neurons based on the brain shown in Figure 1. Posterior view in (A) and anterior view in (A’). The cluster AC (white star in A’) was not analyzed further as it was not Tc-foxQ2II-positive. White circle: tritocerebral cluster; yellow circle: posterior clusters 1 and 2. (B) Schematic overview of the cell body locations for the ten clusters shown in the same colors as in (A/A’) and subsequent figures. White arrowhead: lateral cluster 1 (orange); white arrow: lateral cluster 2 (yellow); open arrowhead: lateral cluster 3 (purple); open arrow: lateral cluster 4 (dark purple); yellow arrowhead: median cluster 1 (magenta); yellow arrow: median cluster 2 (pink); yellow dashed circle: posterior cluster (PC1 is olive green and PC2 is vibrant green); white dashed circle: tritocerebrum cluster (light green and white broken circle in A); white star: anterior cluster (grass yellow). Scale bar: 100 μm.
Figure 3
The morphology and distribution of EGFP-positive clusters.
(A–A’) Posterior view and anterior views of the brain, showing major neuropils. (B–B’) Median cluster 1 (MC1). (B) Posterior view. (B’) Lateral view. The region enclosed by the red dashed square is magnified in the bottom-left corner, with a white arrow indicating the specific projection area. (C–C’) Median cluster 2 (MC2). (C) Posterior view. (C’) Lateral view. The central complex (CX) region, enclosed by a red dashed square, is magnified in the top-right corner. A white arrow indicates projections targeting the VMCpo/PS and LAL. (D–D’) Lateral cluster 1 (LC1). (D) Posterior view. (D’) Anterior view. (E–E’) Posterior view of lateral cluster 2 (LC2). In (E’), a white arrow marks the VMCpo/PS region traversed by LC2. AL: antennal lobe; CB: central body (CBU: upper unit of the central body; CBL: lower unit of the central body); LH: lateral horn; MB: mushroom body (CA: calyx; ML: medial lobe; PED: peduncle; VL: vertical lobe); OL: optic lobe; PB: protocerebral bridge; SLP: superior lateral protocerebrum; SMP: superior medial protocerebrum; TR: tritocerebrum; VMCpo/PS: ventromedial cerebrum postcommissural domain/posterior slope.
Figure 4 with 2 supplements
The morphology and distribution of EGFP-positive clusters in the brain of the foxQ2-5’ line.
(A–A’) Lateral cluster 3 (LC3). (A) Posterior view. (A’) Anterior view. (B–B’) Lateral cluster 4 (LC4). (B) Posterior view. (B’) Lateral view. A white arrow indicates LC4 arborization in the LH and SLP. (C–C’) Posterior cluster 1 (PC1). (C) Posterior view. (C’) Lateral view. The region enclosed by the red dashed square is magnified in the top-right corner. A white arrow shows where PC1 enters the VMCpo/PS. (D–D’) Posterior cluster 2 (PC2). (D) Posterior view. (D’) Lateral view. The projection region, enclosed by a red dashed square, is magnified in the top-right corner. A white arrow points to the arborization site. (E) Anterior view of tritocerebrum cluster (TC). A white arrow indicates its arborization zone. AL: antennal lobe; CB: central body (CBU: upper unit of the central body; CBL: lower unit of the central body); LH: lateral horn; MB: mushroom body (CA: calyx; ML: medial lobe; PED: peduncle; VL: vertical lobe); OL: optic lobe; PB: protocerebral bridge; SLP: superior lateral protocerebrum; SMP: superior medial protocerebrum; TR: tritocerebrum; VMCpo/PS: ventromedial cerebrum postcommissural domain/posterior slope.
Figure 4—figure supplement 1
TC emerges in the tritocerebrum (TR) of the first instar larval (L1) brain.
(A) Larval brain from foxQ2-5’ line stained with EGFP and synapsin antibodies. White arrows in (A) and (B) indicate the EGFP-positive cells located in the TR. (C) highlights the TR component (1), outlined with a red dashed line. Scale bar: 50 μm.
Figure 4—figure supplement 2
EGFP-positive cells without discernible projections.
(A–A’) Posterior view of the brain in the foxQ2-5’ line. The cells indicated by white arrows lacked identifiable projections and were therefore not reconstructed. Scale bar: 100 μm.
Figure 5
Suggested homologous neurons in Drosophila melanogaster.
(A1-A3) Tribolium MC1 mapped to the dopaminergic Drosophila PPM1205 neuron (consistent hemibrain annotation and FAFB prediction). (B1-B3) Tribolium MC2 mapped to PPM1204 in Drosophila (annotated as dopaminergic in hemibraiffn but predicted as glutamatergic in FAFB). (C1-C3) Tribolium LC1a mapped to the dopaminergic PPL101 neuron in Drosophila (consistent hemibrain annotation and FAFB prediction). (D1-E3) Tribolium LC1b tentatively mapped to FB7B (not annotated in hemibrain, predicted as dopaminergic in FAFB) or FB6H (not annotated in hemibrain, predicted as glutamatergic in FAFB) in Drosophila. (F1-F3) Tribolium LC2 mapped to CB0452 in Drosophila (predicted as dopaminergic in FAFB) with soma location matching the PPL2c cluster. (G1-G3) Tribolium LC3 mapped to the dopaminergic Drosophila PPL204 neuron (consistent hemibrain annotation and FAFB prediction). (H1-H3) Tribolium PC1 mapped to DNpe014 in Drosophila (predicted as acetylcholinergic in FAFB). (A-H) Anterior view, lateral view from right of the brain, and the optimal view of each cell type were shown in each panel. (Orientation arrows: A (blue): anterior; D (green): dorsal; R (red): right brain side. ATL: Antler; EB: Ellipsoid body; FB: Fan-shaped body; LAL: Lateral accessory lobe; LH: Lateral horn; LO: Lobula; MB: Mushroom body; PB: Protocerebral bridge; PS: Posterior slope; SMP: Superior medial protocerebrum; SLP: Superior lateral protocerebrum. Scale bars: as indicated.)
Figure 6
Classification of lateral cluster 1 based on distinct projection patterns.
(A–A’) Morphology and projection pattern of LC1a, shown without (A) and with (A’) brain compartments. The region enclosed by the red dashed square shows where LC1a passes near the central body (CB) before crossing the midline. A magnified view of this region (top-right corner) shows the LC1a projection adjacent to the CB, indicated by a white arrow. (B–B’) LC1b projections. The boxed region in (B’) shows that LC1b projects toward the CB. A magnified lateral view (top-right) details the contact point (white arrow). VL: the vertical lobe; ML: the medial lobe; CBU: the upper unit of the central body.
Figure 7 with 5 supplements
Eight clusters detected in the foxQ2-5’-line were also sparsely labeled by Brainbow.
(A) MC1. (B) MC2. (C) LC1. (D) LC2. (E) LC4. (F) PC1. (G) PC2. (H) TC. All panels show the brain of the foxQ2-5’-line. Magenta represents synapsin. Scale bar: 100 μm.
Figure 7—figure supplement 1
Brainbow: stochastic recombination using incompatible lox variants.
(A) In the Brainbow system, Cre randomly selects different excision events (1–3) based on compatible lox sites, respectively. Without Cre activity, no fluorescence signal is detected in the brain due to the presence of a stop codon. Recombination switches expression to mCherry (1), mEYFP (2), or mCerulean (3). (B) In the foxQ2-5’ -line, EGFP fluorescent protein and Cre recombinase are expressed in the same pattern due to an enhancer trap. Since mEYFP and mCerulean fluoresce similarly to EGFP, only mCherry was used as a marker to label cells in the crossed brain. (C) Without Cre activity, RFP is not detected in the Brainbow brain. When the Brainbow line is crossed with the foxQ2-5’ line, recombination activates the expression of mCherry, resulting in RFP being detected in the brain. Scale bar: 100 μm. EFA (3391 bp) is derived from OX466061.1. See Appendix 1 for map and sequence.
Figure 7—figure supplement 2
No notable temperature effect on Cre activity.
Brains grown at 32 °C and 23 °C, stained by immunohistochemistry (IHC). (A–A’) Brain raised at 32 °C stained by IHC. (B–B’) Brain raised at 23 °C stained by IHC. Scale bar: 100 μm.
Figure 7—figure supplement 3
Two LC1 subclusters are sparsely marked with the Brainbow technique.
(A–A’’) LC1a axons bypass the vertical lobe of mushroom bodies and descend in three different brains. (A’’’) Schematic of LC1a projection patterns. (B–B’’) LC1b projects directly to the central body region. (B’’’) Schematic of LC1b projection patterns. Magenta represents synapsin. (n=6). Scale bar: 100 μm.
Figure 7—figure supplement 4
Brainbow marked cells that are not marked by adult Tc-foxQ2 expression.
(A) Columnar neurons (WXYZ tracts) are sparsely labeled by Brainbow. (B1–B5) Some neurons were visualized only once in our analysis. (B6–B7) These neurons were consistently observed in every brain. Scale bar: 100 μm.
Figure 7—figure supplement 5
Brainbow marked cells that are not marked by adult Tc-foxQ2 expression.
Individual cells marked by sparse Brainbow technique. (A1–A7) A cell located in the posterior-dorsal region extends from the vertical lobe (VL) of the mushroom bodies to the anterior-ventral region. (B1–B7) Cells are located in and innervate the antennal lobe. (C1–C5) Cells are located in the ventral part of the brain. (D1–D5) Cells are located in the superior medial protocerebrum (SMP) and project from the dorsal to the ventral part of the brain. (E1–E2) A projection connects the lateral horn (LH) and the antennal lobe (AL). (F1–F2) Cells are located in the medial region of the brain and cross the midline. (G1–G2) Cells are located in the posterior region of the brain and cross the midline. Scale bar: 100 μm.
Figure 8
EGFP-positive cells arborize in the mushroom body and the central complex.
(A–A’’) Arborizations of EGFP-positive cells in the mushroom body (MB). (A) Overview of 3D reconstruction of the MB and EGFP-positive projections, generated using VVDviewer. (A’) Hemi-mushroom body with EGFP-positive projections. (A”) Immunohistochemistry result showing the MB and EGFP-positive projections. White arrows in (A’) and (A’’) indicate EGFP-positive projections arborizing in the α and α’ lobe of the MB. (B–B’’) Arborizations of EGFP-positive cells in the central body (CB). (B) Overview of the 3D reconstruction of the CB with EGFP-positive projections. (B’) Side profile view of the area marked by the white dashed rectangle in (B). (B’’) Immunohistochemistry result showing the CB and EGFP-positive projections. White arrowheads in (B’) and (B’’) indicate EGFP-positive projections arborizing in the upper unit of the central body (CBU). Open arrowheads in (A’) and (B’’) indicate the EGFP-positive projections arborizing in the peduncle (PED). The white dashed circle in (B’’) outlines the PED. Green, EGFP; magenta, synapsin; and white, neuropils (MB or CB).
Figure 9 with 2 supplements
Reconstruction of STED-imaging of LC1a connection of mushroom body with the central complex in the foxQ2-5’-line brain.
(A) LC1a seems to have two main branches: one innervates the mushroom body (MB), and the other innervates the upper unit of the central body (CBU). (B) LC1b arborizes only within the central body (CB). (C) A higher-magnification view of the projections in the CBU (region partially outlined by the white dashed rectangle in A). (D–F) Transverse sections through distinct domains of the central body. (D) Median section. (E) A lateral section showing a single branch projecting into the CB. (F) A section where two distinct tracts are visible. Green and magenta represent the main projections from different sides, which are in close proximity around the midline (white). Yellow indicates arborizations within the CBU. CBU is shown in gray. CBL: lower unit of the central body. The STED dataset used in this panel was acquired by Ying.
Figure 9—figure supplement 1
Tc-foxQ2-dopaminergic neurons of the lateral cluster 1 (LC1) have arborizations in the mushroom body (MB) and the central body (CB).
The arborizations could stem from different neurons, but we cannot rule out that one neuron projects into both neuropils. Red dashed circles indicate components of the MB. PED: peduncle, ML: medial lobe.
Figure 9—figure supplement 2
STED super-resolution microscopy in the Tribolium brain.
(A) Confocal overview of the central brain stained with synapsin (magenta) and Tc-foxQ2-positive neurons (green). Dashed line indicates imaging position in (B), dashed rectangle shows imaging position in (C) and (D). (B–B`) xz-scan (~75 µm in z, FoxQ2 channel only) through the brain in confocal (B) and 3D-STED (B`) mode, showing the resolution increase in 3D-STED. The white arrowhead shows a homogeneous structure in confocal mode, while in 3D-STED four individual neurites can be distinguished. (C) Projection view of a confocal volume. Dashed white line indicates the position of the section in (C`). (C`) Confocal xz-section. White arrowheads indicate an individual FoxQ2-positive neurite (green) running through the neuropil tissue (magenta, synapsin staining). (D) Projection view of a STED (green, FoxQ2-positive neurons) and confocal volume (magenta, synapsin, deconvolved). Dashed white line indicates the position of the section in (D`). (D`) STED xz-section. White arrowheads indicate the same individual FoxQ2-positive neurite shown in (C`) running through the neuropil tissue (magenta, synapsin). A comparison of the neurite in (C`) and (D`) clearly shows the resolution increase in STED compared to the confocal image.
Figure 10 with 6 supplements
A subset of Tc-FoxQ2-positive neurons are dopaminergic.
Co-immunostaining for GFP (green) and the dopamine marker tyrosine hydroxylase (TH, magenta) identifies dopaminergic subtypes among the Tc-FoxQ2-positive neuronal populations. Dashed outlines indicate the brain contour. (A–A’’) The anterior brain region containing lateral cluster 3 (LC3, white arrowheads). (A’) GFP channel. (A’’) TH (dopamine) channel. (B–B’’) A medial section showing the median cluster (MC, open arrowheads), and lateral cluster 2 (LC2, white arrows). (C–C’’) A posterior section showing the median cluster (MC, open arrowheads), lateral cluster 1 (LC1, open arrows), and lateral cluster 2 (LC2, white arrows). Scale bar: 100 μm.
Figure 10—figure supplement 1
GABA is not co-expressed with FoxQ2-positive cells.
(A–A’’) In median cluster (MC), GABA and FoxQ2-positive cells are intermingled in their expression. (B–B’’) Lateral cluster 1 (LC1) shows expression of EGFP only. (C–C’’) In lateral cluster 2 (LC2), no overlap was detected between GABA and FoxQ2 signals. (D–D’’) Lateral cluster 3 (LC3) expresses only EGFP. (E–E’’) In lateral cluster 4 (LC4), GABA and FoxQ2 are not co-expressed in the same cells. (F–F’’) In posterior cluster (PC), GABA and FoxQ2-positive cells are intermingled in their expression. (G–G’’) The tritocerebrum cluster (TC) expresses only EGFP. Scale bar: 20 μm.
Figure 10—figure supplement 2
Serotonin (5-HT) is not co-expressed with FoxQ2-positive cells.
(A–A’’) In median cluster (MC), 5-HT and FoxQ2-positive cells are intermingled in their expression. (B–B’’) Lateral cluster 1 (LC1) shows expression of EGFP only. (C–C’’) In lateral cluster 2 (LC2), no overlap was detected between 5-HT and FoxQ2 signals. (D–D’’) Lateral cluster 3 (LC3) expresses only EGFP. (E–E’’) In lateral cluster 4 (LC4), 5-HT and FoxQ2 are not co-expressed in the same cells. (F–F’’) In posterior cluster (PC), 5-HT and FoxQ2-positive cells are intermingled in their expression. (G–G’’) The tritocerebrum cluster (TC) expresses only EGFP. Scale bar: 20 μm.
Figure 10—figure supplement 3
Choline acetyltransferase (ChAT) is expressed in all FoxQ2-positive subclusters.
(A–A’’) The median cluster (MC), including MC1 and MC2, expresses both ChAT and EGFP. (B–B’’) Lateral cluster 1 (LC1) expresses both ChAT and EGFP. (C–C’’) In lateral cluster 2 (LC2), ChAT and EGFP largely overlap. (D–D’’) Lateral cluster 3 (LC3) expresses both ChAT and EGFP. (E–E’’) In lateral cluster 4 (LC4), ChAT and EGFP largely overlap. (F–F’’) The posterior cluster (PC) expresses both ChAT and EGFP. (G–G’’) The tritocerebrum cluster (TC) expresses both ChAT and EGFP. Scale bar: 20 μm.
Figure 10—figure supplement 4
Glutamate (GLUT) is expressed in all FoxQ2-positive subclusters.
(A–A’’) The median cluster (MC), including MC1 and MC2, expresses both GLUT and EGFP. (B–B’’) In lateral cluster 1 (LC1), GLUT and EGFP largely overlap. (C–C’’) In lateral cluster 2 (LC2), GLUT and EGFP largely overlap. (D–D’’) Lateral cluster 3 (LC3) expresses both GLUT and EGFP. (E–E’’) Lateral cluster 4 (LC4) expresses both GLUT and EGFP. (F–F’’) The posterior cluster (PC) expresses both GLUT and EGFP. (G–G’’) The tritocerebrum cluster (TC) expresses both GLUT and EGFP. Scale bar: 20 μm.
Figure 10—figure supplement 5
Neurotransmitters expressed in the mushroom body and central complex.
(A) Choline acetyltransferase (ChAT). White dashed circles indicate the peduncle (PED) of the mushroom body. The white arrow indicates the signal in the central body. (B) Glutamate (GLUT). White dashed circles represent the peduncle (PED) of the mushroom body. White arrow shows the signals in the central body. (C–C’) Dopamine (TH). White arrows indicate the signals in the α-lobe of the mushroom body. White arrowhead shows the signal in the central body. Scale bar: 100 μm.
Figure 10—figure supplement 6
Octopamine (TBH) is expressed in five FoxQ2-positive subclusters.
(A–A’’) The median cluster (MC), including MC1 and MC2, expresses both TBH and EGFP. (B–B’’) In lateral cluster 1 (LC1), TBH and EGFP largely overlap. (C–C’’) In lateral cluster 2 (LC2), TBH and EGFP largely overlap. (D–D’’) Lateral cluster 3 (LC3) expresses both TBH and EGFP. (E–E’’) Lateral cluster 4 (LC4) expresses only EGFP. (F–F’’) In the posterior cluster (PC), no overlap was detected between TBH and EGFP. (G–G’’) The tritocerebrum cluster (TC) expresses only EGFP. (H) Venn diagrams illustrating the overlap between octopamine-positive and FoxQ2-positive neurons across the four subtypes (n=3). Scale bar: 20 μm.
Figure 11
Co-expression of Tc-foxQ2II and TH (dopamine) in distinct neuronal subsets.
A magnified view highlights neuronal subtypes co-expressing the dopaminergic marker tyrosine hydroxylase (TH) and Tc-FoxQ2/EGFP, alongside an analysis of the signal overlap ratio. (A–D’’) A representative imaging plane showing co-immunostaining for neurons expressing both EGFP (green) and TH (magenta). (E) Schematic of the left-brain hemisphere. Four Tc-FoxQ2-positive neuronal subtypes are outlined with differently colored dashed circles. (F) Venn diagrams quantify the overlap between TH-positive and Tc-FoxQ2-positive neurons for each of the four subtypes (n=5). Scale bar: 20 μm.
Figure 12 with 1 supplement
3D reconstruction of Tc-foxQ2-dopaminergic neuronal projections in the brain of foxQ2-5’-line using VVDviewer.
(A) Overview of the projections from the four neuronal subtypes. (B) Projections from the median cluster (MC). MC1 contains two long tracts (indicated by a white arrowhead), consistent with previous descriptions. (C) Projections from all subtypes within lateral cluster 1 (LC1). (D–D’’’) Individual projections of each LC1 subtype. (D) Overview of the hemi-LC1. (D') LC1a projection (see also Figure 6). (D’’–D’’’) LC1b projections (see also Figure 6). (E) Projections from LC2 and LC3. For the position of the neurons within the brain, see Figures 3, 4 and 6.
Figure 12—figure supplement 1
Reconstruction of dopamine/Tc-foxQ2 double-positive neurons helps to confirm subclusters.
The subset of dopaminergic neurons among EGFP-positive cells was reconstructed using the VVDviewer. (A, B) Overview images of two brains different from the one shown in Figure 12. The median cluster (MC) is shown in magenta; lateral cluster 1 (LC1) in white; lateral cluster 2 (LC2) in yellow; and lateral cluster 3 (LC3) in green. In the MC, the white arrow indicates MC2, while the remaining region corresponds to MC1 (n=2). Only in the sample shown in (A) were the arborizations of LC2 clear enough to reconstruct them.
Appendix 1—figure 1
Map of the Beetle-Brainbow-EFA construct.
Tables
Table 1
Neurotransmitter expression in different EGFP-positive cell types in the foxQ2-5’-line brain.
| Neurotransmitters | Types of EGFP-positive cells in the foxQ2-5’-line | ||||||||
|---|---|---|---|---|---|---|---|---|---|
| MC1 | MC2 | LC1 | LC2 | LC3 | LC4 | PC1 | PC2 | TC | |
| Acetylcholine (ChAT) | |||||||||
| Glutamate | |||||||||
| Dopamine (TH) | |||||||||
| Octopamine (TBH) | |||||||||
| GABA | |||||||||
| Serotonin (5-HT) | |||||||||
-
Color code: gray, positive; white, negative.
-
Gray indicates clusters that contained at least one neuron expressing the respective transmitter. Hence, it does not mean that all cells of a cluster have a respective transmitter nor that transmitters are co-expressed in the neurons.
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Gene (Tribolium castaneum) | Tc-foxQ2II | iBeetle-Base NCBI | TC004761 LOC657858 | Formerly termed Tc-foxQ2 Drosophila ortholog: fd102, CG11152 |
| Strain, strain background (T. castaneum) | San Bernardino | SB | NCBI:txid7070 | |
| Genetic reagent (T. castaneum) | foxQ2-5’-line (generated in the vermilion white strain) | He et al., 2019 | Marks Tc-foxQ2-positive cells with EGFP and Cre; available from the Bucher lab | |
| Antibody | Tc-foxQ2 antibody (guinea pig, polyclonal) | He et al., 2019 | 1:1000 | |
| Antibody | Anti-GFP (chicken, polyclonal) | Abcam | RRID:ab13970 | 1:2000 |
| Antibody | Anti-GFP (rabbit, polyclonal) | Thermo Fisher Scientific | RRID:A11122 | 1:1000 |
| Antibody | Anti-RFP (rabbit polyclonal) | Abcam | RRID:ab62341 | 1:1000 |
| Antibody | Anti-synapsin (3C11; anti SYNORF1) (mouse, monoclonal) | DSHB | RRID:AB_528479 | 1:25 |
| Antibody | Anti-tyrosine-hydroxylase (mouse, monoclonal) | Immunostar | RRID:22941 | 1:1000 |
| Antibody | Anti-serotonin (rabbit, polyclonal) | Sigma-Aldrich, Merck KGaA | RRID:S5545 | 1:1000 |
| Antibody | Anti-GABA (rabbit, polyclonal) | Sigma-Aldrich, Merck KGaA | RRID:A2052 | 1:100 |
| Antibody | Anti-glutamate (mouse, monoclonal) | Sigma-Aldrich | RRID:G9282 | 1:1000 |
| Antibody | Anti-ChAT4B1 (4B1) (mouse, monoclonal) | DSHB | RRID:AB_528122 | 1:200 |
| Antibody | Anti-TβH (rabbit, polyclonal) | Gift from Uwe Homberg | 1:30 | |
| Antibody | Secondary antibodies coupled with Alexa Fluor 488, 555, or 647 (all: goat, polyclonal) | Thermo Fisher Scientific | RRID:A-11039, RRID:A-11070, RRID:A-21435, RRID:A-21425, RRID:A-21430, RRID:A-21450, RRID:A-21235, RRID:A-21245 | 1:1000 |
| Antibody | Anti-rabbit STAR RED (goat, polyclonal) | Abberior | RRID:STRED-1002-500UG | 1:1000 |
| Antibody | Anti-mouse STAR ORANGE (goat, polyclonal) | Abberior | RRID:STORANGE-1001-500UG | 1:1000 |
| Software, algorithm | VVDviewer | Janelia Open Science Software Initiative | Used for reconstructing projections from stacks of stained brains |
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foxQ2 marks fast-acting interneurons including dopaminergic neurons of mushroom bodies and central complex in the beetle Tribolium castaneum
eLife 15:RP111703.
https://doi.org/10.7554/eLife.111703.3
