Selective integration of diverse taste inputs within a single taste modality

  1. Julia U Deere
  2. Arvin A Sarkissian
  3. Meifeng Yang
  4. Hannah A Uttley
  5. Nicole Martinez Santana
  6. Lam Nguyen
  7. Kaushiki Ravi
  8. Anita V Devineni  Is a corresponding author
  1. Zuckerman Mind Brain Behavior Institute, Columbia University, United States
  2. Neuroscience Graduate Program, Emory University, United States
  3. Department of Biology, Emory University, United States
9 figures, 3 tables and 1 additional file

Figures

Models for bitter taste processing and Gal4 lines to target bitter neuron subsets.

(A) Schematic depicting three major taste organs in the fly. (B) Models for how different subsets of bitter-sensing neurons could be processed in the brain. (C–D) Expression patterns of Gal4 lines …

Figure 1—source data 1

Expression pattern of Gal4 lines used to target bitter neuron subsets.

https://cdn.elifesciences.org/articles/84856/elife-84856-fig1-data1-v1.xlsx
Figure 2 with 1 supplement
Bitter neuron subsets in different organs act in parallel to regulate feeding responses.

(A) Effect of activating bitter neuron subsets on proboscis extension response (PER) to 100 mM sucrose (n=5–8 sets of flies). Bar graphs for each neuronal subset (left) represent the fraction of …

Figure 2—figure supplement 1
Additional characterization of the effects of bitter neuron manipulations on feeding.

(A) Schematic of the microstructure of fly feeding behavior, as defined in Itskov et al., 2014. Feeding activity bouts are periods of time when the fly is frequently interacting with the food, while …

Figure 3 with 2 supplements
Bitter neuron subsets in three different organs elicit similar effects on locomotion.

(A) Arena used for tracking flies during optogenetic activation. (B) Schematic of protocol to test the effect of bitter neuron activation on locomotion. 5 s light stimulation was used at three …

Figure 3—figure supplement 1
Additional characterization of locomotor effects elicited by activating all bitter neurons.

(A) Fraction of flies moving when all bitter neurons were activated using Gr33a-Gal4 (n=11–12 trials). Traces show behavior with medium intensity light stimulation. Bar graphs show change in …

Figure 3—figure supplement 2
Additional characterization of locomotor effects elicited by subsets of bitter neurons.

(A) Change in forward speed (top) or turning (bottom) elicited by low or high intensity light stimulation (n=10–12 trials). Results for medium intensity stimulation are shown in Figure 3E–F. (B) …

Figure 4 with 2 supplements
Subsets of bitter neurons in multiple organs elicit innate and learned aversion.

(A) Protocol to test the effect of bitter neuron activation on innate positional aversion. (B–C) Effect of activating all bitter-sensing neurons (B) or subsets of bitter neurons (C) on innate …

Figure 4—figure supplement 1
Additional characterization of innate aversion elicited by activating all bitter neurons.

(A) Innate positional aversion elicited by the activation of all bitter-sensing neurons at medium or high light intensity (n=22–24 trials, 11–12 sets of flies). Traces for low light intensity and …

Figure 4—figure supplement 2
Effects of bitter neuron subsets on innate positional aversion at other light intensities.

Graphs show the preference index (PI) during the last 5 s of light presentation at medium (A) or high (B) light intensity (n=20–24 trials, 10–12 sets of flies). The effect of low intensity light is …

Summary of behavioral effects elicited by optogenetic activation of bitter neuron subsets.

(A) Effects are color-coded by strength, relative to the effect of activating all bitter neurons with Gr33a-Gal4. All observed effects went in the same direction. See Materials and methods for …

Figure 5—source data 1

Summary of behavioral effects elicited by optogenetic activation of bitter neuron subsets.

https://cdn.elifesciences.org/articles/84856/elife-84856-fig5-data1-v1.xlsx
Figure 6 with 1 supplement
Bitter inputs from different organs are relayed to overlapping downstream pathways.

(A) The entire population of second-order bitter neurons was labeled by trans-Tango tracing from Gr33a-Gal4-expressing cells. Arrows denote the three major tracts (lateral, mediolateral, and medial) …

Figure 6—figure supplement 1
Additional images of trans-Tango labeling.

Additional images of trans-Tango labeling, with presynaptic bitter-sensing neurons labeled in green and postsynaptic second-order neurons labeled in red. The top row represents the same brains shown …

Figure 7 with 2 supplements
Second-order mlSEZt bitter neurons receive input from multiple organs.

(A) Top: R29F12-Gal4 expression pattern (maximum intensity projection created from images generated by the Janelia FlyLight Project Team). Brain slices at the far anterior and posterior edges are …

Figure 7—figure supplement 1
Further characterization of mlSEZt responses.

(A) Image of R29F12-Gal4 expression (same image as Figure 7A) showing the approximate location of subesophageal zone (SEZ) and superior lateral protocerebrum (SLP) imaging examples in panels B and …

Figure 7—figure supplement 2
Responses of mlSEZt cells labeled by R55E01-Gal4 and R29F12-R55E01 split-Gal4.

(A) R55E01-Gal4 expression pattern (maximum intensity projection created from images generated by the Janelia FlyLight Project Team). Brain slices at the far anterior and posterior edges are not …

Figure 8 with 3 supplements
Second-order mlSEZt neurons regulate a subset of taste-related behaviors.

(A) Expression pattern of split-Gal4 line (R29F12-AD+R55E01-DBD) labeling mlSEZt neurons with high specificity. Pink arrowhead shows mlSEZt cell bodies; white and yellow arrows denote their …

Figure 8—figure supplement 1
Additional characterization of mlSEZt effects on feeding behavior.

(A–D) Additional characterization of the effects of activating (A–B; n=42–47 flies) or silencing (C–D; n=66–69 flies) mlSEZt neurons. These data are derived from the same experiments shown in Figure …

Figure 8—figure supplement 2
Additional characterization of mlSEZt effects on locomotor and preference behaviors.

(A–D) mlSEZt neurons were activated using R29F12-R55E01 split-Gal4 driving UAS-Chrimson. (A, C) Effects of low and medium intensity light stimulation on forward velocity (A) or turning (C) (n=11–12 …

Figure 8—figure supplement 3
Effects of silencing mlSEZt neurons on locomotor and preference behaviors elicited by bitter neuron activation.

(A–D) Effects of activating bitter-sensing neurons (Gr66a-lexA driving Aop-Chrim) while silencing mlSEZt neurons (mlSEZt split-Gal4 driving UAS-Kir). Flies lacking Aop-Chrim were used as a negative …

Figure 9 with 2 supplements
Downstream pathways from mlSEZt cells.

(A) Colabeling of mlSEZt cells with markers for GABAergic (top) or cholinergic (bottom) neurons. Each row shows mlSEZt labeling (red), the neurotransmitter marker (green), and both channels …

Figure 9—figure supplement 1
Morphology of mlSEZt neurons in male brains.

Examples of mlSEZt cells in male brains labeled with the split-Gal4 line (R29F12-AD+R55E01-DBD). No reproducible differences in morphology were identified between mlSEZt neurons in male and female …

Figure 9—figure supplement 2
Additional images of mlSEZt colabeling with neurotransmitter markers.

(A–B) Additional images showing colabeling of mlSEZt cells with markers for GABAergic (A) or cholinergic (B) neurons. Each column shows mlSEZt labeling (red), the neurotransmitter marker (green), …

Tables

Table 1
List of 3Ns receiving at least 20 synapses from mlSEZt cells.

Percent input from mlSEZt refers to the percent of total input synapses the cell receives that come from mlSEZt cells. Target regions for each cell refer to areas where the cell has output synapses. …

3N cell typeCell ID# mlSEZt input cellsmlSEZt input types# mlSEZt synapses% Input from mlSEZtTarget regions
SLP1914209735997mAL3A, mAL3B646.6AVLP, SCL, SIP, SLP
SLP1323598926696mAL3A, mAL4542.1AVLP, LH, SCL, SIP, SLP
SMP3895751974828mAL3A, mAL3B,mAL4483.4AVLP, PLP, SCL, SIP, SLP, SMP
SLP1914213135638mAL3A, mAL3B, mAL4475.0AVLP, LH, SCL, SIP, SLP
aSP-g3B4216509825mAL3A4612.9AVLP, PLP, SCL, SIP, SLP, SMP, SMP(C)
LHAV4l13602367244mAL4413.3AVLP, LH, SCL, SIP, SLP
SLP179_b4206238735mAL3B, mAL4383.7SCL, SIP, SLP
LHAV2f2_b5747101215mAL3A, mAL3B, mAL4387.9AVLP, LH, SCL, SLP
LHAV2f2_a6047355254mAL3B, mAL4369.4AVLP, LH, SCL, SLP
LHAV2f2_b5733463245mAL3A, mAL4355.5AVLP, LH, SCL, SLP
aSP-g3A3299190365mAL3A, mAL3B, mAL4356.3SIP, SLP, SMP(C), SMP
LHAV1e13902710334mAL3A, mAL3B, mAL4352.2AVLP, LH, SCL, SIP, SLP
SLP015_e3933404024mAL3B, mAL4336.0SIP, SLP
SLP0112975197367mAL3A, mAL4332.2LH, SCL, SIP, SLP
SLP179_b3913111865mAL3B, mAL4323.3SIP, SLP
SLP015_c3592401445mAL3B, mAL4283.1AVLP, SCL, SIP, SLP
aSP-g3B4854303365mAL3A279.2AVLP, SCL, SIP, SLP, SMP
SLP1875785219412mAL3B2711.8SIP, SLP
SLP1876078209372mAL3B267.3AVLP, SCL, SLP
SLP3762982543844mAL3B, mAL4250.6SIP, SLP, SMP
SLP21658130111197mAL3A, mAL3B, mAL4251.0AOTU, AVLP, GOR, GOR(C), IB, ICL, LH, PLP, SCL, SIP, SLP, SMP, SMP(C)
SLP2156085340973mAL3B, mAL4241.1AVLP, PLP, SCL, SLP, VES
SLP015_c3599269234mAL3B, mAL4235.4SIP, SLP
SLP1874837118113mAL3B238.9SLP
SLP25958130407074mAL3B, mAL4231.7AVLP, SCL, SIP, SLP, SMP
AVLP0244209651173mAL3B221.2AVLP, LH, PVLP, SCL, SIP, SLP
LHCENT13288612824mAL4220.2AVLP, LH, MB, SCL, SIP, SLP, SMP
LHAV6a1058130472553mAL4226.5LH, SIP, SLP
SLP015_e3292066282mAL4213.2SIP, SLP
SMP5504526894943mAL3B, mAL4210.4AVLP, LH, PLP, SCL, SIP, SLP, SMP
SLP0184516631724mAL3A, mAL3B, mAL4203.1AVLP, LH, SCL, SIP, SLP
SLP05758130199554mAL3B, mAL4200.5AVLP, LH, PLP, SCL, SIP, SLP
Table 2
List of top 30 4Ns.

Top 30 4Ns were selected and sorted based on the total number of synapses from top 3Ns. Percent input from 3Ns refers to the percent of total input synapses the cell receives that come from the top …

4N cell typeCell ID# 3N input cells3N input cell types# synapses from 3Ns% input from 3NsTarget regions
SMP54829758058916AVLP024, LHAV1e1, SLP015_c, SLP015_e, SLP057, SLP179_b, SLP191, SLP216, SMP389, SMP550, aSP-g3A, aSP-g3B3636.3AVLP, LH, PLP, SCL, SIP, SLP, SMP
SLP2793605918608LHAV1e1, LHCENT1, SLP015_e, SLP018, SLP179_b, SLP216, SMP550, aSP-g3A3548.4SCL, SIP, SLP, SMP, SMP(C)
LHCENT45175062654LHAV4l1, LHCENT1, SLP057, SMP3892974.6AVLP, LH,, MB, PLP, SCL, SIP, SLP, SMP, mALT
DM1_lPN5426348181LHCENT12472.4AL, LH, MB, SCL, SLP, mALT
LHMB158130209882LHCENT1, SLP0572383.5CRE, LH, MB, SCL, SIP, SLP, SMP
SLP38829825861111AVLP024, SLP015_e, SLP018, SLP179_b, SLP191, SLP216, SMP389, aSP-g3B2033.1SCL, SIP, SLP, SMP, SMP(C)
SMP5504526894947AVLP024, LHAV1e1, SLP018, SLP216, SMP389, aSP-g3B2014.2AVLP, LH, PLP, SCL, SIP, SLP, SMP
oviIN4231011892SMP389, SMP5501900.8CAN, CRE, CRE(C), GOR, IB, LAL, SIP, SIP(C), SMP, SMP(C), SPS, VES
LHCENT933026894010AVLP024, LHAV1e1, LHCENT1, SLP132, SLP191, SMP389, aSP-g3A, aSP-g3B1771.4AOTU, AVLP, LH, MB, SCL, SIP, SLP, SMP
SLP212581298052914AVLP024, SLP015_e, SLP018, SLP057, SLP132, SLP191, SLP216, SMP389, SMP550, aSP-g3A, aSP-g3B1456.7SIP, SLP, SMP, SMP(C)
SMP1082982585133SLP057, SMP389, SMP5501450.6CRE, CRE(C), LAL, LH, MB, MB(C), SCL, SIP(C), SIP, SLP, SMP, SMP(C)
oviDNa5506556682SLP216, SMP55013712.3CRE, SCL, SIP, SLP, SMP, SMP(C), VES
MBON1858130208281LHCENT11321.0LH, MB, SCL, SIP, SLP
LHPD4c14216418599LHAV1e1, LHAV4l1, LHCENT1, SLP011, SLP018, SLP057, SLP132, SLP1871302.5LH, SCL, SIP, SLP, SMP
SLP1133905895916AVLP024, LHAV2f2_b, LHAV4l1, LHCENT1, SLP13212710.5LH, SIP, SLP, SMP
LHPV10b16047097272LHCENT1, SLP0571252.8CRE, LH, MB, PLP, PVLP, SCL, SIP, SLP, SMP
SMP1566737767693SLP216, SMP389, SMP5501242.1CRE, GOR(C), IB, ICL(C), ICL, LAL, MB, SMP, SMP(C), SPS, SPS(C)
SMP38558130837802SMP389, SMP5501242.0CRE, CRE(C), LAL, LAL(C), SCL, SIP, SIP(C), SMP, SMP(C),
SLP44032887047213LHAV1e1, SLP015_c, SLP015_e, SLP018, SLP057, SLP179_b, SLP191, SLP376, aSP-g3B1233.0LH, SCL, SIP, SLP, SMP, SMP(C)
LHPV5e13286110045LHAV1e1, LHAV4l1, LHCENT1, SLP015_e, SLP1321231.1CRE, CRE(C), LH, MB(C), SCL, SIP, SIP(C), SLP, SMP, SMP(C)
SMP0296040704333SLP216, SMP389, SMP5501206.7AVLP, LH, MB, PLP, SCL, SIP, SLP, SMP
SMP31158130493782SMP389, SMP5501177.5AVLP, ICL, PLP, SCL, SIP, SLP, SMP
PPL2013285337616LHAV1e1, LHCENT1, SLP011, SLP015_e, SLP057, SLP179_b1171.4AVLP, CRE, LH, MB, PLP, POC, SCL, SIP, SLP, SMP, mALT
SMP10958130096202SMP389, SMP5501071.7AOTU, CRE, CRE(C), LAL, LAL(C), MB, SIP, SMP, VES
SMP0295413478114SLP215, SLP216, SMP389, SMP5501065.8AVLP, MB, PLP, SCL, SIP, SLP, SMP
SMP50336131280812AVLP024, LHCENT1, SLP011, SLP015_e, SLP018, SLP132, SLP187, SLP215, SMP389, SMP550, aSP-g3B1021.5AVLP, CRE, LH, MB, PLP, SCL, SIP, SIP(C), SLP, SMP, SMP(C)
SLP1134212713058LHAV2f2_a, LHAV2f2_b, LHAV4l1, LHCENT1, SLP132, SLP18710210.0LH, SIP, SLP, SMP, SMP(C)
MBON184571964441LHCENT1973.6LH, MB(C), SCL, SIP, SIP(C), SLP
SLP14958130093129LHAV1e1, LHAV4l1, LHCENT1, SLP015_c, SLP018, SLP057, SLP179_b, SLP376974.3SIP, SLP, SMP
SLP441581307854213LHAV1e1, LHCENT1, SLP011, SLP015_c, SLP015_e, SLP057, SLP179_b, SLP191, SLP259, SLP376, aSP-g3B963.3AVLP, SCL, SIP, SLP, SMP
Key resources table
Reagent type (species) or resourceDesignationSource or referenceIdentifiersAdditional information
Genetic reagent (Drosophila melanogaster)Gr33a-Gal4Moon et al., 2009BDSC: 31425
Genetic reagent (Drosophila melanogaster)Gr58c-Gal4Weiss et al., 2011BDSC: 57646
Genetic reagent (Drosophila melanogaster)Gr59c-Gal4Weiss et al., 2011BDSC: 57650
Genetic reagent (Drosophila melanogaster)Gr22f-Gal4Weiss et al., 2011BDSC: 57610
Genetic reagent (Drosophila melanogaster)Gr9a-Gal4Weiss et al., 2011BDSC: 57596
Genetic reagent (Drosophila melanogaster)R29F12-Gal4Jenett et al., 2012BDSC: 49495
Genetic reagent (Drosophila melanogaster)R55E01-Gal4Jenett et al., 2012BDSC: 39117
Genetic reagent (Drosophila melanogaster)R29F12-ADDionne et al., 2018BDSC: 71164
Genetic reagent (Drosophila melanogaster)R55E01-DBDDionne et al., 2018BDSC: 69662
Genetic reagent (Drosophila melanogaster)Gr66a-lexAThistle et al., 2012BDSC: 93024
Genetic reagent (Drosophila melanogaster)VGAT-lexADeng et al., 2019BDSC: 84441
Genetic reagent (Drosophila melanogaster)ChAT-lexADeng et al., 2019BDSC: 84379
Genetic reagent (Drosophila melanogaster)UAS-Chrimson-TdTDuistermars et al., 2018N/A
Genetic reagent (Drosophila melanogaster)UAS-GTACR1-TdTB NoroN/A
Genetic reagent (Drosophila melanogaster)UAS-Kir2.1Baines et al., 2001BDSC: 6595
Genetic reagent (Drosophila melanogaster)trans-Tango reporter (UAS-Myr-GFP, QUAS-mtdTomato; trans-Tango)Talay et al., 2017BDSC: 77124
Genetic reagent (Drosophila melanogaster)UAS-GCaMP6fChen et al., 2013BDSC: 42747
Genetic reagent (Drosophila melanogaster)UAS-TdTVK5D HattoriN/A
Genetic reagent (Drosophila melanogaster)UAS-TdTp40G Rubin and B PfeifferBDSC: 32222
Genetic reagent (Drosophila melanogaster)lexAop-GCaMP6fD Kim; Hattori et al., 2017BDSC 44277
AntibodyAnti-GFP (chicken polyclonal)Aves LabsCat# GFP-1020; RRID: AB_100002401:1000
AntibodyAnti-DsRed (rabbit polyclonal)ClontechCat# 632496; RRID: AB_100134831:500
AntibodyAnti-bruchpilot (nc82; mouse monoclonal)Development Studies Hybridoma BankCat# nc82; RRID: AB_23148661:10
AntibodyAlexa Fluor 488 (goat anti-chicken polyclonal)Life TechnologiesCat# A11039; RRID: AB_25340961:500
AntibodyAlexa Fluor 568 (goat anti-rabbit polyclonal)Life TechnologiesCat# A11036; RRID: AB_105635661:500
AntibodyAlexa Fluor 633 (goat anti-mouse polyclonal)Life TechnologiesCat# A21052; RRID: AB_25357191:500
Software, algorithmPrism, version 9GraphPadN/A
Software, algorithmMATLABMathworksN/A
Software, algorithmFlyTrackerCaltech; Eyjolfsdottir et al., 2014N/Ahttp://www.vision.caltech.edu/Tools/FlyTracker/

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