Transsynaptic mapping of Drosophila mushroom body output neurons

  1. Kristin M Scaplen  Is a corresponding author
  2. Mustafa Talay
  3. John D Fisher
  4. Raphael Cohn
  5. Altar Sorkaç
  6. Yoshi Aso
  7. Gilad Barnea  Is a corresponding author
  8. Karla R Kaun  Is a corresponding author
  1. Department of Neuroscience, Brown University, United States
  2. Department of Psychology, Bryant University, United States
  3. Center for Health and Behavioral Sciences, Bryant University, United States
  4. Laboratory of Neurophysiology and Behavior, The Rockefeller University, United States
  5. Janelia Research Campus, Howard Hughes Medical Institute, United States
8 figures, 1 table and 2 additional files

Figures

Figure 1 with 23 supplements
MBONs have divergent connections across the brain.

Exemplar max-stacks of glutamatergic MBONs (A) MB011B, (B) MB002B, (C) MB399B, (D) MB310C, (E) MB434B, (F) MB298B, GABAergic MBONs (G) MB110C and (H) MB057B, and cholinergic MBONs (I) MB077B, (J) MB018B, (K) MB026B, (L) MB080C, (M) MB082C, (N) MB542B, (O) MB050B, (P) MB051C, (Q) MB549C and (R) MB027B, trans-Tango identified postsynaptic connections. For max-stacks: green, presynaptic MBONs, magenta, postsynaptic trans-Tango signal, blue, brp-SNAP neuropil. A map of the MBONs that are included in the expression pattern in each driver line accompanies each exemplar with the relative expression pattern (grayscale, 1–5) accordingly to FlyLight (https://splitgal4.janelia.org/cgi-bin/splitgal4.cgi). MBON maps are organized by neurotransmitter type: green=glutamatergic, blue=GABAergic, red=cholinergic. Scale bar = 50 μm.

Figure 1—figure supplement 1
MBON driver lines that have similar expression patterns also have similar postsynaptic connections across the brain.

Exemplar max-stacks of glutamatergic MBONs (A) MB074C, (B) MB210B, (C) MB433B, (D) GABAergic MBONs MB083C, and cholinergic MBONs (E) MB051B and (F) MB077C, trans-Tango identified postsynaptic connections. For max-stacks: green, presynaptic MBONs, magenta, postsynaptic trans-Tango signal, blue, brp-SNAP neuropil. A map of the MBONs that are included in the expression pattern in each driver line accompanies each exemplar with the relative expression pattern (grayscale, 1–5) accordingly to FlyLight (https://splitgal4.janelia.org/cgi-bin/splitgal4.cgi). MBON maps are organized by neurotransmitter type: green=glutamatergic, blue=GABAergic, red=cholinergic. Scale bar = 50 μm.

Figure 1—figure supplement 2
Full-size exemplar max-stack of MB011B.

Green, presynaptic MBONs, magenta, postsynaptic trans-Tango signal, blue, brp-SNAP neuropil. Scale bar = 50 μm.

Figure 1—figure supplement 3
Full-size exemplar max-stack of MB002B.

Green, presynaptic MBONs, magenta, postsynaptic trans-Tango signal, blue, brp-SNAP neuropil. Scale bar = 50 μm.

Figure 1—figure supplement 4
Full-size exemplar max-stack of MB399B.

Green, presynaptic MBONs, magenta, postsynaptic trans-Tango signal, blue, brp-SNAP neuropil. Scale bar = 50 μm.

Figure 1—figure supplement 5
Full-size exemplar max-stack of MB310C.

Green, presynaptic MBONs, magenta, postsynaptic trans-Tango signal, blue, brp-SNAP neuropil. Scale bar = 50 μm.

Figure 1—figure supplement 6
Full-size exemplar max-stack of MB434B.

Green, presynaptic MBONs, magenta, postsynaptic trans-Tango signal, blue, brp-SNAP neuropil. Scale bar = 50 μm.

Figure 1—figure supplement 7
Full-size exemplar max-stack of MB298B.

Green, presynaptic MBONs, magenta, postsynaptic trans-Tango signal, blue, brp-SNAP neuropil. Scale bar = 50 μm.

Figure 1—figure supplement 8
Full-size exemplar max-stack of MB110C.

Green, presynaptic MBONs, magenta, postsynaptic trans-Tango signal, blue, brp-SNAP neuropil. Scale bar = 50 μm.

Figure 1—figure supplement 9
Full-size exemplar max-stack of MB057B.

Green, presynaptic MBONs, magenta, postsynaptic trans-Tango signal, blue, brp-SNAP neuropil. Scale bar = 50 μm.

Figure 1—figure supplement 10
Full-size exemplar max-stack of MB077B.

Green, presynaptic MBONs, magenta, postsynaptic trans-Tango signal, blue, brp-SNAP neuropil. Scale bar = 50 μm.

Figure 1—figure supplement 11
Full-size exemplar max-stack of MB018B.

Green, presynaptic MBONs, magenta, postsynaptic trans-Tango signal, blue, brp-SNAP neuropil. Scale bar = 50 μm.

Figure 1—figure supplement 12
Full-size exemplar max-stack of MB026B.

Green, presynaptic MBONs, magenta, postsynaptic trans-Tango signal, blue, brp-SNAP neuropil. Scale bar = 50 μm.

Figure 1—figure supplement 13
Full-size exemplar max-stack of MB080C.

Green, presynaptic MBONs, magenta, postsynaptic trans-Tango signal, blue, brp-SNAP neuropil. Scale bar = 50 μm.

Figure 1—figure supplement 14
Full-size exemplar max-stack of MB082C.

Green, presynaptic MBONs, magenta, postsynaptic trans-Tango signal, blue, brp-SNAP neuropil. Scale bar = 50 μm.

Figure 1—figure supplement 15
Full-size exemplar max-stack of MB051C.

Green, presynaptic MBONs, magenta, postsynaptic trans-Tango signal, blue, brp-SNAP neuropil. Scale bar = 50 μm.

Figure 1—figure supplement 16
Full-size exemplar max-stack of MB549C.

Green, presynaptic MBONs, magenta, postsynaptic trans-Tango signal, blue, brp-SNAP neuropil. Scale bar = 50 μm.

Figure 1—figure supplement 17
Full-size exemplar max-stack of MB027B.

Green, presynaptic MBONs, magenta, postsynaptic trans-Tango signal, blue, brp-SNAP neuropil. Scale bar = 50 μm.

Figure 1—figure supplement 18
Full-size exemplar max-stack of MB074C.

Green, presynaptic MBONs, magenta, postsynaptic trans-Tango signal, blue, brp-SNAP neuropil. Scale bar = 50 μm.

Figure 1—figure supplement 19
Full-size exemplar max-stack of MB210B.

Green, presynaptic MBONs, magenta, postsynaptic trans-Tango signal, blue, brp-SNAP neuropil. Scale bar = 50 μm.

Figure 1—figure supplement 20
Full-size exemplar max-stack of MB433B.

Green, presynaptic MBONs, magenta, postsynaptic trans-Tango signal, blue, brp-SNAP neuropil. Scale bar = 50 μm.

Figure 1—figure supplement 21
Full-size exemplar max-stack of MB083C.

Green, presynaptic MBONs, magenta, postsynaptic trans-Tango signal, blue, brp-SNAP neuropil. Scale bar = 50 μm.

Figure 1—figure supplement 22
Full-size exemplar max-stack of MB051B.

Green, presynaptic MBONs, magenta, postsynaptic trans-Tango signal, blue, brp-SNAP neuropil. Scale bar = 50 μm.

Figure 1—figure supplement 23
Full-size exemplar max-stack of MB077B.

Green, presynaptic MBONs, magenta, postsynaptic trans-Tango signal, blue, brp-SNAP neuropil. Scale bar = 50 μm.

Figure 2 with 1 supplement
Whole brain distribution of MBON postsynaptic connections overlap.

(A) Example of presynaptic MBON γ5β′2a (SS01308) and postsynaptic trans-Tango signal in a registered brain. For max-stacks: green, presynaptic MBONs, magenta, postsynaptic trans-Tango signal. (B) Example of segmented trans-tango signals that was continuous to MBON γ5β′2a terminals. For max-stack: gray, postsynaptic trans-Tango signal. (C) Heatmap displaying the overlap in segmented MBON postsynaptic signal by brain region. Postsynaptic signal for each MBON was normalized within each brain to capture respective expression levels. SS01308 was used to target MBON γ5β′2a, MB399B was used to target MBON β2β′2a, MB002B was used to target MBONs γ5β′2a, β′2mp, SS01143 was used to target MBON β′2mp, MB011B was used to target MBONs γ5β′2a, β′2mp, β′2mp_bi, MB057B was used to target MBON β′1, and MB110C was used to target MBONs γ3, γ3β′1. MB433B was used to target MBONs β1>α, γ4>γ1γ2, MB298B was used to target MBON γ4>γ1γ2, MB077C was used to target MBON γ2α′1 and MB50B was used to target MBONs α′1, α2sc. MB018B was used to target MBON α′2, MB027B was used to target MBON α′3ap, α′3 m, and SS01194 was used to target MBON α2sc. For raw postsynaptic signal see Figure 2—figure supplement 1. (D) Schematic of fly brain highlighting the most anterior brain regions included in mask analysis starting at AL and ending with SLP. (E) Schematic of fly brain highlighting the most posterior brain regions included in mask analysis starting at NO and ending with PB. AL: antennal lobe, AMMC: antennal mechanosensory and motor center, ATL: antler, AVLP: anterior ventrolateral protocerebrum, CRE: crepine, EB: ellipsoid body, EPA: epaulette, FSB: fan-shaped body, FLA: flange, GA: shoulder of lateral accessory lobe, GOR: gorget of ventral complex, IB: interior bridge, ICL: inferior clamp, IPS: inferior posterior slope, IVLP: inferior ventrolateral protocerebrum, LAL: lateral accessory lobe, LB: bulb of lateral complex, LH: lateral horn, MB: mushroom body, NO: noduli, OTU: optic tubercle, PB: protocerebral bridge, PLP: posterior lateral protocerebrum, PRW: prow, PVLP: posterior ventrolateral protocerebrum, SAD: saddle, SCL: superior clamp, SEG: subesophageal ganglion, SIP: superior intermediate protocerebrum, SLP: superior lateral protocerebrum, SMP: superior medial protocerebrum, SPS: superior posterior plate, VES: vest of ventral complex, WED: wedge. Scale bar = 50 μm.

Figure 2—figure supplement 1
Whole brain distribution of MBON postsynaptic connections.

Heatmap displaying the unscaled overlap in signal of segmented MBON postsynaptic signal by brain region.

Figure 3 with 2 supplements
DANs postsynaptic to MBONs.

Exemplar max-stacks of MBON lines in which TH+ cells overlapped with postsynaptic signal of glutamatergic (A) MBON γ5β′2a, β′2mp, β′2mp_bilateral (MB011B), (B) MBON γ5β′2a, β′2mp (MB002B), (C) MBON γ5β′2a, β′2mp, β2β′2a (MB074C), (D) GABAergic MBONs γ3, γ3β′1 (MB083C) and (E) cholinergic MBONs γ2α′1 (MB077C). Overlapping TH+ and trans-Tango cell bodies are highlighted in insets, scale bar = 10 μm. Max stacks of MB are included (Column I), scale bar = 50 μm. Column II-IV depict single optical planes from anterior to posterior outlining MB compartments. Bar graphs indicate the average number of co-localized cells per hemibrain (mean +/- standard error). Green, TH-positive cells; magenta, postsynaptic trans-Tango signal. MBON maps are organized by neurotransmitter type: green=glutamatergic, blue=GABAergic, red=cholinergic. (F) Schematic depicting the MB innervation by PAM DANs. PAM DANs extend dendrites to SMP, CRE, SIP, and SLP. (G) Schematic depicting the MBONs that synapse on TH+ cells.

Figure 3—figure supplement 1
DANs postsynaptic to MBONs.

Exemplar max-stacks of MBON lines in which TH+ cells were not overlapping or had few overlapping cells with postsynaptic signal of glutamatergic (A) MBON α1 (MB310C), (B) MBON γ4>γ1 γ2, β1>α (MB433B), (C) MBON γ4>γ1 γ2 (MB298B), (D) GABAergic MBON β′1 (MB057B), (E) cholinergic MBON α′3 (MB027B), (F) MBON α2sc (MB080C), and (G) MBONs α′1, α2p3p, α′3m (MB542B). Max stacks of MB are included, scale bar = 50 μm. Bar graphs indicate the average number of co-localized cells per hemibrain (mean ± standard error). Green, TH-positive cells; magenta, postsynaptic trans-Tango signal. MBON maps are organized by neurotransmitter type: green=glutamatergic, blue=GABAergic, red=cholinergic.

Figure 3—figure supplement 2
Total PAM TH+ cells counted.

Bar graphs indicate the average number of PAM TH+ cells counted per hemibrain (mean ± standard error).

Figure 4 with 1 supplement
Subsets of MBONs converge on other MBONs.

(A) MBON β′2mp receives convergent input from glutamatergic MBON γ5β′2a (MB011B), GABAergic MBONs γ3, γ3β′1 (MB110C) and cholinergic MBON γ2α′1 (MB077B) and MBON α′2 (MB018B). (B) MBON γ3β′1 receives convergent input from glutamatergic MBON β′2mp (MB002B) and MBON γ4>γ1γ2 (MB298B). β′2mp, γ3 and β′1 are outlined in representative stacks. (C) Schematics summarizing identified convergent MBONs (β′2mp and γ3β′1) and their respective convergent input. Solid lines represent the convergent MBON and dotted lines represent convergent input. For max-stacks: green, presynaptic MBONs, magenta, postsynaptic trans-Tango signal, blue, brp-SNAP neuropil, scale bar=50 μm.

Figure 4—figure supplement 1
Patterns of MBON convergence is consistent across MBON driver lines that have similar MBON expression.

MBON γ3β′1 receives input from (A) glutamatergic MBON β′2mp (MB074C). MBON β′2mp receives convergent input from (B) glutamatergic MBON γ5β′2a (MB210B), (C) GABAergic MBONs γ3, γ3β′1 (MB083C) and (D) cholinergic MBON γ2α′1 (MB051C) and (E) MBON α′2 (MB082C). For max-stacks: green, presynaptic MBONs, magenta, postsynaptic trans-Tango signal, blue, brp-SNAP neuropil, scale bar=50 μm.

Figure 5 with 2 supplements
MBONs converge on different layers of the FSB.

Exemplar max-stacks of glutamatergic (A), GABAergic (B), and cholinergic (C) MBONs whose postsynaptic neurons innervate the FSB. Max-stacks are approximately 50 μm thick. Slices were selected based on the relative position of the FSB. For FSB stacks: magenta, postsynaptic trans-Tango signal, blue, brp-SNAP neuropil. Map of MBONs accompany each exemplar with the relative expression pattern (grayscale, 1–5) accordingly to FlyLight. For each map, green=glutamatergic, blue=GABAergic, red=cholinergic. Scale bar = 50 μm. (D) Map summarizing the percentage of trans-Tango-positive signal in each FSB layer across brains for each MBON. (E) Schematic depicting MBONs that converge onto different layers of the FSB. MB compartments are colorized based on the neurotransmitter expressed by the MBON that innervates it. Lines thickness corresponds to the percentage of trans-Tango-positive signal in each FSB layer across brains for each MBON.

Figure 5—figure supplement 1
MBON α1 postsynaptic signal innervating FSB in females.

Exemplar max-stacks of glutamatergic MBON α1 whose postsynaptic neurons innervate the FSB from two female flies highlighting the broad FSB innervation pattern of postsynaptic signa. Max-stacks are approximately 50 μm thick. Magenta, postsynaptic trans-Tango signal; blue, neuropil. Scale bar = 50 μm.

Figure 5—figure supplement 2
Variability in FSB postsynaptic signal.

Exemplar max-stacks of cholinergic MBON α`2 whose postsynaptic neurons innervate the FSB highlighting the variability that existed in FSB innervation across eight different brains. Max-stacks are approximately 50 μm thick. Magenta, postsynaptic trans-Tango signal; blue, neuropil. Scale bar = 50 μm.

MBONs converge onto LAL neurons.

Exemplar max-stacks of glutamatergic (A), GABAergic (B), and cholinergic (C) MBONs innervating the LAL. Max-stacks are approximately 50 μm thick. Slices were selected based on the relative position of the LAL. Magenta, postsynaptic trans-Tango signal, blue, brp-SNAP neuropil. Map of MBONs accompany each exemplar with the relative expression pattern (grayscale, 1–5) accordingly to FlyLight. For each map green=glutamatergic, blue=GABAergic, red=cholinergic. Scale bar = 50 μm. Scale bar for insets = 10 μm (D) Map summarizing the percentage of trans-Tango-positive signal in LAL across brains for each MBON. (E) Schematic depicting MBONs that converge onto neurons innervating the LAL. MB compartments are colorized based on the neurotransmitter expressed by the MBON that innervates it. Lines thickness corresponds to the percentage of trans-Tango-positive signal in LAL across brains for each MBON.

Figure 7 with 3 supplements
MBONs γ3β′1 and γ2α′1 converge onto the same subset of LAL and FSB neurons.

Exemplar max-stacks of cholinergic MBON γ2α′1 (MB077C) postsynaptic connections and identified overlap with respective (A) FSB (47H09) and (B) LAL (VT015539). (C) Confirmation of functional connection with optogenetic activation of MB077C and calcium imaging of FSB neurons in SMP and FSB (47H09), and calcium imaging of LAL neurons in SMP (VT015539). The red bar indicates when the LED was on and the shutter was closed to protect the PMTs during LED stimulation. Exemplar max-stacks of GABAergic MBON γ3β′1 (MB083C) postsynaptic connections and identified overlap with respective (D) FSB (47H09) and (E) LAL (VT015539). Max-stacks are approximately 50 μm thick. Slices were selected based on the relative position of the LAL and FSB. In A, B, D and E, red, postsynaptic trans-Tango signal; blue, CD2 marker of split-GAL4 line; green, LexA FSB or LAL. Scale bar = 50 μm. (F) Schematic highlighting convergence of MBONs γ3β′1 and γ2α′1 onto the same genetically identified subsets of LAL and FSB neurons. (G) Max-stack of SS32219; green, GFP expression; blue, neuropil. Scale bar = 50 μm. (H) shibirets (shits) inactivation of LAL using split-GAL4 SS32219 resulted in significant increases in group activity (F(2,21)=39.28 p<0.0001). Group activity counts were binned over 10 s periods, averaged across biological replicates of 10 flies each (n = 8) and plotted against time. Lines depict mean +/- standard error. (I) One video was selected at random of each genotype and processed using FlyTracker to calculate the average pathlength (F(2,29)=33.39, p<0.0001), angular velocity (F(2,29)=51.87, p<0.0001) and velocity (F(2,29)=30.97, p<0.0001) of individual flies. Box plots with overlaid raw data were generated using RStudio. Each dot is a single fly. One-way ANOVA with Tukey Posthoc was used to compare mean and variance. ***p<0.0001.

Figure 7—figure supplement 1
Optogenetic activation of MBON α2sc (MB080C) does not result in changes in signal recorded from of LAL neurons in SMP (VT015539).
Figure 7—figure supplement 2
Inactivation of LAL using split-GAL4 SS32230 results in significant increases in group activity.

(A) Max-stack of SS32230; green, GFP expression; blue, neuropil. Scale bar = 50 μm. (B) shits inactivation of LAL using split-GAL4 SS32230 resulted in significant increases in group activity (F(2,21)=13.94 p<0.0001). Group activity counts were binned over 10s periods, averaged across biological replicates of 10 flies each (n = 8) and plotted against time. Lines depict mean ± standard error. (C) One video was selected at random of each genotype and processed using FlyTracker to calculate the average pathlength (F(2,28)=14.76, p<0.0001), angular velocity (F(2,28)=28.16, p<0.0001), and velocity (F(2,28)=13.96, p<0.0001) of individual flies. Box plots with overlaid raw data were generated using RStudio. Each dot is a single fly. One-way ANOVA with Tukey Posthoc was used to compare mean and variance. ***p<0.0001, **p<0.001.

Figure 7—figure supplement 3
Group activity of split-GAL4 SS32219 and SS32230 at permission temperatures.

Group activity of experimental groups were not significantly different from both genetic control groups. (A) Average group activity was significantly different at permissive temperatures for split-GAL4 line SS32219 (F(2,21)=4.617, p=0.02) although Tukey Post-hoc analysis revealed that shits/SS32219 was not significantly different from either genetic control (SS32219/+ vs shits/SS32219, p=0.08, shits/+ vs shits/SS32219, p=0.82). Instead, shits/+ was significantly different from SS32219/+ (p=0.02). (B) Average group activity was significantly different at permissive temperatures for split-GAL4 line SS32230 (F(2,21)=6.195, p=0.0078) although Tukey Post-hoc analysis revealed that shits/SS32230 was not significantly different from both genetic controls (SS32230/+ vs shits/ SS32230, p=0.97, shits/+ vs shits/ SS32230, p=0.013). Instead, SS32230/+ was significantly different from both shits/ SS32230 (p=0.013) and shits/+ (p=0.02).

Summary schematics highlighting postsynaptic connections of MBON innervating (A) innervating the protocerebrum (B) PAM DANs (solid lines) and MBONs (dotted lines).

(C) FSB and LAL. Lines thickness corresponds to the percentage of trans-Tango-positive signal in FSB and LAL across brains for each MBON. (D) Schematic highlighting convergence of MBONs γ3β′1 and γ2α′1 onto the same genetically identified subsets of LAL and FSB neurons (solid lines). Dotted lines depict the established connections between the FSB and LAL (Wolff and Strausfeld, 2015b).

Tables

Key resources table
Reagent type
(species) or
resource
DesignationSource or
reference
IdentifiersAdditional
information
Genetic reagent (D. melanogaster)y[1]w[*]Pfeiffer et al., 2008
Genetic reagent (D. melanogaster)UAS-shibirets1Pfeiffer et al., 2012FLYB: FBst0066600; RRID:BDSC_66600
Genetic reagent (D. melanogaster)LexAop-GCaMP6s, UAS-ChrimsonAllan Wong (Janelia Research Campus)N/A13xLexAop2-Syn21-opGCaMP6s in su(Hw)attP8, 10xUAS-Syn21-Chrimson88-tdTomato-3.1 in attP18
Genetic reagent (D. melanogaster)trans-TangoTalay et al., 2017FLYB: FBst0077124;
RRID:BDSC_ 77124
trans-Tango in attP40
Genetic reagent (D. melanogaster)UAS-myrGFP, QUAS-mtdTomatoTalay et al., 2017FLYB: FBst0077479; RRID:BDSC_7747910xUAS-myrGFP, 5xQUAS-mtdTomato(3xHA) in su(Hw)attP8
Genetic reagent (D. melanogaster)UAS-CD2, QUAS-mtdTomatoThis studyN/A10xUAS-CD2, 5xQUAS-mtdTomato(3xHA) in su(Hw)attP8
Genetic reagent (D. melanogaster)brp-SNAPKohl et al., 2014FLYB: FBst0058397;
RRID:BDSC_ 58397
brp[SNAPf-tag]/Cyo
Genetic reagent (D. melanogaster)LexAop-GFPPfeiffer et al., 2010FLYB: FBst0032203;
RRID:BDSC_32203
13XLexAop2-mCD8::GFP in attP2
Genetic reagent (D. melanogaster)MB002B-split-GAL4Aso et al., 2014aFlyLight Robot ID: 2135053
RRID:BDSC_68305
MBON β′2mp (4), γ5β′2a (2)
Genetic reagent (D. melanogaster)MB011B-split-GAL4Aso et al., 2014aFlyLight Robot ID: 2135062
RRID:BDSC_68294
MBON γ5β′2a (4), β′2mp (3), β′2mp_bilateral (3)
Genetic reagent (D. melanogaster)MB018B-split-GAL4Aso et al., 2014aFlyLight Robot ID: 2135069
RRID:BDSC_68296
MBON α′2 (4)
Genetic reagent (D. melanogaster)MB026B-split-GAL4Aso et al., 2014aFlyLight Robot ID: 2135077
RRID:BDSC_68300
MBON α′1 (3), α′3ap (2)
Genetic reagent (D. melanogaster)MB027B-split-GAL4Aso et al., 2014aFlyLight Robot ID: 2135078
RRID:BDSC_68301
MBON α′3ap (5), α′3 m (5)
Genetic reagent (D. melanogaster)MB050B-split-GAL4Aso et al., 2014aFlyLight Robot ID: 2135100
RRID:BDSC_68365
MBON α′1 (2), α2sc (4)
Genetic reagent (D. melanogaster)MB051B-split-GAL4Aso et al., 2014aFlyLight Robot ID: 2135101
RRID:BDSC_68275
MBON α′2 (1), γ2α′1 (4)
Genetic reagent (D. melanogaster)MB051C-split-GAL4Aso et al., 2014aFlyLight Robot ID: 2135136
RRID:BDSC_68249
MBON α′2 (1), γ2α′1 (3)
Genetic reagent (D. melanogaster)MB057B-split-GAL4Aso et al., 2014aFlyLight Robot ID: 2135106
RRID:BDSC_68277
MBON β′1 (3)
Genetic reagent (D. melanogaster)MB074C-split-GAL4Aso et al., 2014aFlyLight Robot ID: 2135122
RRID:BDSC_68282
MBON β′2mp (4), β2β′2a (3), γ5β′2a (1)
Genetic reagent (D. melanogaster)MB077B- split-GAL4Aso et al., 2014aRRID:BDSC_68283MBON γ2α′1 (4)
Genetic reagent (D. melanogaster)MB077C- split-GAL4Aso et al., 2014aFlyLight Robot ID: 2135125
RRID:BDSC_68284
MBON γ2α′1 (3)
Genetic reagent (D. melanogaster)MB080C- split-GAL4Aso et al., 2014aFlyLight Robot ID: 2135128
RRID:BDSC_68285
MBON α2sc (2)
Genetic reagent (D. melanogaster)MB082C- split-GAL4Aso et al., 2014aFlyLight Robot ID: 2135130
RRID:BDSC_68286
MBON α′2 (3), α3 (5)
Genetic reagent (D. melanogaster)MB083C- split-GAL4Aso et al., 2014aFlyLight Robot ID: 2135131
RRID:BDSC_68287
MBON γ3 (5), γ3β′1 (5)
Genetic reagent (D. melanogaster)MB093C- split-GAL4Aso et al., 2014aFlyLight Robot ID: 2135141
RRID:BDSC_68289
MBON α′2 (4)
Genetic reagent (D. melanogaster)MB110C-split-GAL4Aso et al., 2014aFlyLight Robot ID: 2135158
RRID:BDSC_68262
MBON γ3 (5), γ3β′1 (5)
Genetic reagent (D. melanogaster)MB210B-split-GAL4Aso et al., 2014aFlyLight Robot ID: 2135258
RRID:BDSC_68272
MBON γ5β′2a (1), β′2mp (4), β2β′2a (3)
Genetic reagent (D. melanogaster)MB298B-split-GAL4Aso et al., 2014aFlyLight Robot ID: 2135346
RRID:BDSC_68309
MBON γ4>γ1γ2 (4)
Genetic reagent (D. melanogaster)MB310C-split-GAL4Aso et al., 2014aFlyLight Robot ID: 2135358
RRID:BDSC_68313
MBON α1 (5)
Genetic reagent (D. melanogaster)MB399B-split-GAL4Aso et al., 2014aFlyLight Robot ID: 2501738
RRID:BDSC_68369
MBON β2β′2a (2)
Genetic reagent (D. melanogaster)MB433B-split-GAL4Aso et al., 2014aFlyLight Robot ID: 2501774
RRID:BDSC_68324
MBON β1>α (3), γ4>γ1γ2 (4)
Genetic reagent (D. melanogaster)MB434B-split-GAL4Aso et al., 2014aFlyLight Robot ID: 2501775
RRID:BDSC_68325
MBON β1>α (4), γ4>γ1γ2 (4)
Genetic reagent (D. melanogaster)MB542B-split-GAL4Aso et al., 2014aFlyLight Robot ID: 2501887
RRID:BDSC_68372
MBON α′1 (1), α′3 m (2), α2p3p (2)
Genetic reagent (D. melanogaster)GMR47H09-LexAPfeiffer et al., 2013FLY: FBtp0088666
RRID:BDSC_53482
Genetic reagent (D. melanogaster)VT055139-LexATirian and Dickson, 2017N/A
Genetic reagent (D. melanogaster)VT018476-lexABidaye et al., 2014N/A
Genetic reagent (D. melanogaster)SS01308-split GAL4Janelia Research CampusN/AMBON γ5β′2a
Genetic reagent (D. melanogaster) SS01143-split GAL4Janelia Research CampusN/AMBON β′2mp
Genetic reagent (D. melanogaster)SS1194-split GAL4Janelia Research CampusN/AMBON α2sc
Genetic reagent (D. melanogaster)SS32219-split GAL4Janelia Research CampusN/ALateral Accessory Lobe
Genetic reagent (D. melanogaster)SS32230-split GAL4Janelia Research CampusN/ALateral Accessory Lobe
Antibodyα-GFP (Rabbit polyclonal)Life TechCat #A11122
RRID:AB_221569
(1:1000)
Antibodyα-HA (Rat monoclonal)RocheCat #11867423001
RRID:AB_390918
(1:100)
Antibodyα-GFP (Chicken polyclonal)ClontechCat #ab13970
RRID:AB_300798
(1:2000)
Antibodyα-DS (Rabbit monoclonal)ClontechCat #632496
RRID:AB_10013483
(1:1000)
Antibodyα-CD2 (Mouse monoclonal)Bio-RadCat #MCA154GA
RRID:AB_566608
(1:100)
Antibodyα-TH (Mouse monoclonal)ImmunostarCat #22941
RRID:AB_572268
(1:500)
AntibodyGoat α-Mouse AF647 (polyclonal)Thermo FisherCat #A21235
RRID:AB_2535804
(1:1000)
AntibodyGoat α-Rabbit AF488
(polyclonal)
Life TechCat #A11034
RRID:AB_2576217
(1:400)
AntibodyGoat α-Rat AF568 (polyclonal)Life TechCat #A11077
RRID:AB_2534121
(1:400)
AntibodyGoat α-Chicken
AF488
(polyclonal)
Life TechCat #A11039
RRID:AB_2534096
(1:400)
AntibodyGoat α-Rabbit AF568
(polyclonal)
Life TechCat #A11011
RRID:AB_143157
(1:400)
SoftwareAdobe Illustrator CCAdobeRRID:SCR_014199
SoftwareZENCarl Zeiss MicroscopyVersion 2.1 (blue edition)
RRID:SCR_013672
SoftwareFijihttp://fiji.scRRID:SCR_002285

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  1. Kristin M Scaplen
  2. Mustafa Talay
  3. John D Fisher
  4. Raphael Cohn
  5. Altar Sorkaç
  6. Yoshi Aso
  7. Gilad Barnea
  8. Karla R Kaun
(2021)
Transsynaptic mapping of Drosophila mushroom body output neurons
eLife 10:e63379.
https://doi.org/10.7554/eLife.63379