DOI: 10.5061/dryad.ncjsxkt25
title: Data from: Glutamate sensor and calcium signals in dopamine neurons and dopamine release

description: We recorded glutamate input to dopamine neurons in the ventral tegmental
 area (VTA) with iGluSnFR(A184S), a glutamate sensor, somatodendritic
 dopamine neuron activity in the VTA with jGCaMP7f, a calcium indicator,
 and dopamine release at the ventral striatum (VS) with GrabDA2m, a
 dopamine sensor, using fiber fluorometry. Recordings were performed in two
 types of reward association tasks (classical conditioning and sequential
 conditioning), with presentation of aversive airpuff to the eye with or
 without buprenorphine treatment, and with optogenetic activation of
 glutamate input axons at the VTA.
description: <strong>Behavior</strong> After 1
 week of recovery from surgery, mice were water-restricted in their cages.
 All conditioning tasks were controlled by a NIDAQ board and LabVIEW. Mice
 were handled for 2 days, acclimated to the experimental setup for 1-2 days
 including consumption of water from the tube, and head-fixed with random
 interval water for 1-3 days until mice showed reliable water consumption.
 For odor-based classical conditioning, all mice were head-fixed, and the
 volume of water reward was constant for all reward trials (predicted or
 unpredicted) in all conditions (6 ml). Some sessions included mild air
 puff trials, directed at one of the eyes and the intensity of air puff was
 constant for all air puff trials (predicted or unpredicted; 2.5 psi). In
 classical conditioning, each association trial began with an odor cue
 (lasting 1 s) followed by a 2 s delay, and then an outcome (either water,
 nothing, or air puff) was delivered. In sequential conditioning, each
 association trial began with a distal odor cue (lasting 1 s) followed by a
 2 s delay, and then a distal odor cue (lasting 1 s) followed by 1 s delay,
 and then an outcome (either water, or nothing) was delivered. Some trial
 types began with a proximal odor cue (lasting 1 s) followed by 1 s delay,
 and then an outcome (either water, or nothing) was delivered. Odors were
 delivered using a custom olfactometer. Each odor was dissolved in mineral
 oil at 1:10 dilution and 30 ml of diluted odor solution was applied to the
 syringe filter (2.7mm pore, 13mm; Whatman, 6823-1327). Odorized air was
 further diluted with filtered air by 1:8 to produce a 900 ml/min total
 flow rate. Different sets of odors (Ethyl butyrate, p-Cymene, Isoamyl
 acetate, Isobutyl propionate, 1-Butanol, 4-Methylanisole, Caproic acid,
 Eugenol, and 1-Hexanol) were selected for each animal. Some of the animals
 shared the same odor set (4 animals and 3 animals for DA sensor classical
 conditioning; 3 animals and 2 animals for sequential
 conditioning) A variable inter-trial interval (ITI) of
 flat hazard function (minimum 10s, mean 13s, truncated at 20s) was placed
 between trials. Each session was composed of multiple blocks (12-24
 trial/block) and all trial types were pseudorandomized in each block. Each
 day, the mice did about 70-350 trials over the course of 20-75 min, and
 with constant excitation from the laser and continuous recording in
 recording sessions. Training for classical conditioning
 used 4 types of trials; odor cue predicting 100% water, odor cue
 predicting 40% water/60% no outcome (nothing), odor cue predicting nothing
 (29.4% of all trials for each odor), and water without cue (free water)
 (11.8%) for 7-10 days, and then odor cue predicting 80% water/20% nothing,
 odor cue predicting 40% water/60% nothing, odor cue predicting nothing
 (29.4% each), and free water (11.8%) for more than 2 days of training
 followed by recording sessions. The first 170 trials or trials before the
 animal stops licking for each session are used for analysis.  3 animals
 used for glutamate sensor recording (Figure 2) were trained with classical
 conditioning with air puff trial; odor cue predicting 100% water, odor cue
 predicting 100% air puff, odor cue predicting 40% water/60% no outcome
 (nothing), odor cue predicting nothing (20.8 % of all trials for each
 odor), and water without cue (free water), air puff without cue (free air
 puff)  (8.3 % of all trials for each stimulus) for 8-9 days, and then odor
 cue predicting 80% water/20% nothing, odor cue predicting 80% air puff/20%
 nothing, odor cue predicting 40% water/60% nothing, odor cue predicting
 nothing (20.8 % each), and water without cue (free water), air puff
 without cue (free air puff) (8.3% each) for more than 7 days of training
 followed by recording sessions. The first 192 trials or trials before the
 animal stops licking for each session are used for analysis. Of note these
 3 animals are used only in the analysis for glutamate sensor activity
 pattern, and not included for comparison between different sensor
 recordings. For sequential conditioning step1, mice
 were trained with proximal odor for 3-7 days, using 3 types of trials;
 proximal odor cue predicting 100% water, proximal odor cue predicting
 nothing (45.8 % of all trials for each odor), and water without cue (free
 water) (8.3 %). Then, for sequential conditioning step2, mice were further
 trained with distal odor and proximal odor using 6 types of trials; distal
 odor cue predicting proximal odor cue predicting 100% water (25 %), distal
 odor cue predicting 50% proximal odor cue predicting water/50% proximal
 odor cue predicting nothing (25 %), distal odor cue predicting proximal
 odor cue predicting nothing (25 %), proximal odor cue predicting 100%
 water (8.3 %), proximal odor cue predicting nothing (8.3 %), and water
 without cue (free water) (8.3 %). Sensor signals were recorded after 8-12
 days of step2 training. The first 192 trials of each session are used for
 analysis. For buprenorphine treatment,<strong>
 </strong>the mouse received unexpected air puff and water with a
 variable ITI of the flat hazard function (minimum 10s, mean 13s, truncated
 at 20s). Animals were first acclimated to head-fixation with water reward
 for 2 days. Then the animals were habituated to a test procedure for two
 days, which was composed of 40-70 trials of air puff and water presented
 in pseudorandomized order (the order of trials is pseudorandomized within
 each block of 10 trials composed of the same number of air puff trials and
 reward trials) with subcutaneous injection of saline (10 ml/g in body
 weights) at the end of sessions. After habituation sessions, buprenorphine
 sessions and control saline sessions were performed in pseudorandom order.
 Each session was separated into pre-injection sub-session and
 post-injection sub-session. The sub-session was composed of 48-75 trials
 with the air puff and water trials in the pseudorandom order. After
 pre-injection sub-sessions, either buprenorphine (Buprenorphine
 hydrochloride, Par Pharmaceutical: diluted in saline 0.03 mg/ml) or
 control saline was subcutaneously injected (10 ml/g; 0.3 mg/g
 buprenorphine or corresponding volume of saline). Post-injection
 sub-session started 5 min after injection. To prevent acute adverse
 effects, we used a smaller dose of buprenorphine (0.15 mg/g) in the first
 session and this session is not included in the analysis.
 <strong>Optogenetic stimulation</strong>
 Red light from 625 nm LED light (M625F2, Thorlab) was applied
 through an optical patch cord (400 mm, 0.39 NA, Thorlab). 12 mW single
 block pulse light of 20, 50, and 250ms duration was triggered pseudorandom
 order through custom software written in LabVIEW (National Instruments)
 via a NIDAQ board (PCI-e6321, National Instruments). A variable
 inter-trial interval (ITI) of flat hazard function (minimum 10s, mean 13s,
 truncated at 20s) was placed between trials.
 <strong>Data analysis</strong>
 Fiber-fluorometry The noise from the power
 line in the voltage signal was cleaned by removing 58-62Hz signals through
 a band stop filter. The global change within a session was normalized
 using a moving median of 100 s. Then, the correlation between green and
 red signals during ITI was examined by linear regression when the red
 signal data was available. If the correlation is significant (<em>p
 </em>≤ 0.05), fitted tdTomato signals were subtracted from green
 signals. Responses were calculated by subtracting the average baseline
 activity from the average activity of the target window. Z-scores of the
 signals were obtained using mean and standard deviation of signals in all
 ITI (from 5 s before odor onset to odor onset for classical conditioning,
 from 5 s before distal odor onset to odor onset for sequential
 conditioning) in each animal. Licking
 Licking from a water spout was detected by a photoelectric sensor
 that produces a change in voltage when the light path is broken. The
 timing of each lick was detected at the peak of the voltage signal above a
 threshold.
description: # Glutamate sensor and calcium signals in dopamine neurons and dopamine
 release The recording data for glutamate input to dopamine,
 somatodendritic dopamine activity, and dopamine release at the ventral
 striatum. The datasets are divided into 5 different task conditions:
 ‘Classical_conditioning’, ‘Sequential conditioning’, ‘Airpuff_response’,
 ‘Airpuff_with_Buprenorphine’, and ‘Optogenetic_stimulation’. ##
 Description of the data and file structure **File names** Data of each
 recording session is stored in each “.mat” file (**_********.mat). Task
 type is indicated at the beginning of the file name (‘task
 type’******.mat). The mouse ID number is indicated in the middle of the
 file name (**_***’mouse ID’***.mat). The session number is indicated at
 the end of the file name (**_**_**_session’session number’.mat). Task
 structures were exactly the same in each session, and each session was
 performed on different days. The block name is indicated at the end of the
 file name after the session number in some files (see below). \** **
 **Classical_conditioning** **Files start from “CC_Glu_”** Activity of
 glutamate inputs to dopamine neurons in VTA was recorded using a glutamate
 sensor, SF-iGluSnFR(A184S), during classical conditioning tasks. Animals
 were trained for more than nine days before neural recording.
 “dFoverFGCaMP2” is the motion-corrected  and z-scored glutamate signal
 (see Methods for the details). Each row corresponds to each trial (-1 sec
 to 8 sec from water onset for free reward trial, and -1 sec to 8 sec from
 odor onset for other trial types) in the order in the session, and each
 column corresponds to signals in 1 ms. “lickmatrix” is the lick event
 stamp (1: lick, 0: no lick) for the same time window as photometry data.
 “TrialType” is the list of trial types in the session. Trial types for the
 task are as follows. Mouse ID: 18, 436, 455, 516, 524, 537, 541, 577, 578,
 580, 581, 587 1: free reward trial (reward at 1001) 9: 80%
 reward-predicting cue followed by reward (cue at 1001 and reward at 4001)
 10: 80% reward-predicting cue followed by reward omission (cue at 1001)
 11: 40% reward-predicting cue followed by reward (cue at 1001 and reward
 at 4001) 12: 40% reward-predicting cue followed by reward omission (cue at
 1001) 14: 0% reward (nothing)-predicting cue (cue at 1001)   Mouse ID:
 346, 362, 364 1: free reward trial (reward at 1001) 4: free airpuff trial
 (airpuff at 1001) 5: 80% airpuff predicting cue followed by airpuff (cue
 at 1001 and airpuff at 4001) 9: 80% reward predicting cue followed by
 reward (cue at 1001 and reward at 4001) 10: 80% reward predicting cue
 followed by reward omission (cue at 1001) 11: 40% reward predicting cue
 followed by reward (cue at 1001 and reward at 4001) 12: 40% reward
 predicting cue followed by reward omission (cue at 1001) 14: 0% reward
 (nothing) predicting cue (cue at 1001) 16: 80% airpuff predicting cue
 followed by airpuff omission (cue at 1001)   **Files start from “CC_DA_”**
 Dopamine sensor signal data recorded from VS neurons during classical
 conditioning are stored in these files. “dFoverFGCaMP2” is the z-scored
 dopamine sensor signal for each trial (-1 sec to 8 sec from water onset
 for free reward trial, and -1 sec to 8 sec from odor onset for other trial
 types) in a single session. “lickmatrix” is the lick event stamp (1: lick,
 0: no lick) for the same time window as photometry data. “TrialType” is
 the list of trial type order in the session. Trial types for the task are
 as follows. All mice 1: free reward trial (reward at 1001) 9: 80% reward
 predicting cue reward (cue at 1001 and reward at 4001) 10: 80% reward
 predicting cue followed by reward omission (cue at 1001) 11: 40% reward
 predicting cue followed by reward (cue at 1001 and reward at 4001) 12: 40%
 reward predicting cue followed by reward omission (cue at 1001) 14: 0%
 reward (nothing) predicting cue (cue at 1001)   **Files start from
 “CC_GCaMP_”** Calcium sensor signal data recorded from VTA dopamine
 neurons during classical conditioning are stored in these files.
 “dFoverFGCaMP2” is the z-scored dopamine sensor signal for each trial (-1
 sec to 8 sec from water onset for free reward trial, and -1 sec to 8 sec
 from odor onset for other trial types) in a single session. “lickmatrix”
 is the lick event stamp (1: lick, 0: no lick) for the same time window as
 photometry data. “TrialType” is the list of trial type order in the
 session. Trial types for the task are as follows. All mice 1: Free reward
 trial (reward at 1001) 9: 80% reward predicting cue followed by reward
 (cue at 1001 and reward at 4001) 10: 80% reward predicting cue followed by
 reward omission (cue at 1001) 11: 40% reward predicting cue followed by
 reward (cue at 1001 and reward at 4001) 12: 40% reward predicting cue
 followed by reward omission (cue at 1001) 14: 0% reward (nothing)
 predicting cue (cue at 1001)   **Sequential_conditioning** **Files start
 from “SeqC_GluDA_”** Simultaneously recorded glutamate sensor signals from
 VS neurons and dopamine sensor signals from VTA dopamine neurons during
 sequential conditioning are stored in these files.   “dFoverFGCaMP2” is
 the motion-corrected and z-scored glutamate sensor signal, and
 “dFoverFGCaMP” is the z-scored dopamine sensor signal for each trial in a
 single session. -6 sec to 8 sec from water onset for free reward trial
 (trial type 1), -4sec to 10 sec from proximal cue onset for unexpected
 proximal cue trial (trial types 9 and 14), and -1 sec to 13 sec from
 distal odor onset for other trial types (trial types 18, 19, 20, and 21).
 “lickmatrix” is the lick event stamp (1: lick, 0: no lick) for the same
 time window as photometry data. “TrialType” is the list of trial type
 order in the session. Trial types for the task are as follows. All mice 1:
 Free reward trial (reward at 6001) 9: Unexpected 100% reward predicting
 proximal cue at 4001 and reward at 6001 14: Unexpected 0% reward (nothing)
 predicting proximal cue at 4001 18: Distal cue 100% predicting 100% reward
 predicting proximal cue at 1001/ 100% reward predicting proximal cue at
 4001/ reward at 6001 19: Distal cue 50% predicting 100% reward or 0%
 reward predicting proximal cue at 1001/ 100% reward predicting proximal
 cue at 4001/ reward at 6001 20: Distal cue 50% predicting 100% reward or
 0% reward predicting proximal cue at 1001/ 0% reward predicting proximal
 cue at 4001 21: Distal cue 100% predicting 0% reward (nothing) predicting
 proximal cue at 1001/ 0% reward predicting proximal cue at 4001  
 **Airpuff_response** **Files start from “Ap_GluDA_”** Mouse ID: 361, 363,
 457, 524, and 537 Simultaneously recorded glutamate sensor signals from
 VTA dopamine neurons and dopamine sensor signals from VS neurons during
 free airpuff and free reward trials are stored in these files.
 “dFoverFGCaMP2” is the z-scored and motion-corrected glutamate signal, and
 “dFoverFGCaMP” is the z-scored dopamine sensor signal for each trial in a
 single session. -1 sec to 8 sec from water or airpuff onset. “lickmatrix”
 is the lick event stamp (1: lick, 0: no lick) for the same time window as
 photometry data. “TrialType” is the list of trial type order in the
 session. Trial types for the task are as follows. Mouse ID: 457 and 537 1:
 Free reward trial (reward at 1001) 4: Free airpuff trial (airpuff at 1001)
   Mouse ID: 361, 363, and 524 1: Free reward trial (reward at 1001) 4:
 Free airpuff trial (airpuff at 1001) 5: 80% airpuff predicting cue
 followed by airpuff (cue at 1001 and airpuff at 4001) 9: 80% reward
 predicting cue followed by reward (cue at 1001 and reward at 4001) 10: 80%
 reward predicting cue followed by reward omission (cue at 1001) 11: 40%
 reward predicting cue followed by reward (cue at 1001 and reward at 4001)
 12: 40% reward predicting cue followed by reward omission (cue at 1001)
 14: 0% reward (nothing) predicting cue (cue at 1001) 16: 80% airpuff
 predicting cue followed by airpuff omission (cue at 1001)   **Files start
 from “Ap_Glu_”** Mouse ID: 455, 578, and 587 Glutamate sensor signals from
 VTA dopamine neurons at free airpuff and free reward trials are stored in
 these files. “dFoverFGCaMP2” is the z-scored and motion-corrected
 glutamate signal for each trial in a single session. -1 sec to 8 sec from
 water or airpuff onset. “lickmatrix” is the lick event stamp (1: lick, 0:
 no lick) for the same time window as photometry data. “TrialType” is the
 list of trial type order in the session. Trial types for the task are as
 follows. 1: Free reward trial (reward at 1001) 4: Free airpuff trial
 (airpuff at 1001)   **Files start from “Ap_DA_”** Mouse ID: 362 and 456
 Dopamine sensor signals from VS neurons during free airpuff and free
 reward trials are stored in these files. “dFoverFGCaMP2” is the z-scored
 dopamine signal for each trial in a single session. -1 sec to 8 sec from
 water or airpuff onset. “lickmatrix” is the lick event stamp (1: lick, 0:
 no lick) for the same time window as photometry data. “TrialType” is the
 list of trial type order in the session. Trial types for the task are as
 follows. 1: Free reward trial (reward at 1001) 4: Free airpuff trial
 (airpuff at 1001)   **Files start from “Ap_GCaMP_”** Mouse ID: 582, 583,
 585, 586, 588, 597, 599, 600, and 602 Calcium sensor signal data during
 free airpuff and free reward trials are stored in these files.
 “dFoverFGCaMP2” is the z-scored dopamine signal for each trial in a single
 session. -1 sec to 8 sec from water or airpuff onset. “lickmatrix” is the
 lick event stamp (1: lick, 0: no lick) for the same time window as
 photometry data. “TrialType” is the list of trial type order in the
 session. Trial types for the task are as follows. 1: Free reward trial
 (reward at 1001) 4: Free airpuff trial (airpuff at 1001)  
 **Airpuff_with_Buprenorphine** (Correspond to Figure 8) **Files start from
 “ApOpi_GluDA_”** Simultaneously recorded glutamate sensor signals from VTA
 dopamine neurons and dopamine sensor signals from VS neurons for free
 airpuff and free reward responses before and after buprenorphine treatment
 are stored in these files. The treatment (“_pre”: before buprenorphine
 injection, and “_post”: after buprenorphine treatment) is indicated at the
 end of the file name. “dFoverFGCaMP2” is the z-scored and motion-corrected
 glutamate signal, and “dFoverFGCaMP” is the z-scored dopamine sensor
 signal for each trial in a single session. -1 sec to 8 sec from water or
 airpuff onset. “lickmatrix” is the lick event stamp (1: lick, 0: no lick)
 for the same time window as photometry data. “TrialType” is the list of
 trial type order in the session. Trial types for the task are as follows.
 All mice 1: Free reward trial (reward at 1001) 4: Free airpuff trial
 (airpuff at 1001)   **Files start from “ApSal_GluDA_”** Simultaneously
 recorded glutamate sensor signal and dopamine sensor signal data for free
 airpuff and free reward responses before and after control saline
 treatment are stored in these files. The treatment (“_pre”: before saline
 injection, and “_post”: after saline injection) is indicated at the end of
 the file name. “dFoverFGCaMP2” is the z-scored and motion-corrected
 glutamate signal, and “dFoverFGCaMP” is the z-scored dopamine sensor
 signal for each trial in a single session. -1 sec to 8 sec from water or
 airpuff onset. “lickmatrix” is the lick event stamp (1: lick, 0: no lick)
 for the same time window as photometry data. “TrialType” is the list of
 trial type order in the session. Trial types for the task are as follows.
 All mice 1: Free reward trial (reward at 1001) 4: Free airpuff trial
 (airpuff at 1001)   **Optogenetic_stimulation** (Correspond to Figure 1)
 **Files start from “OptoChr_Glu_”** Mouse ID: 590, 606, 613, and 614
 Glutamate sensor signal data during free light stimulation (single 20, 50,
 or 250 ms pulse) and free reward trials in the mice expressing ChrimsonR
 in glutamate input to the VTA are stored in these files. “dFoverFGCaMP2”
 is the z-scored glutamate signal for each trial in a single session. -1
 sec to 8 sec from water or airpuff onset. “lickmatrix” is the lick event
 stamp (1: lick, 0: no lick) for the same time window as photometry data.
 “TrialType” is the list of trial type order in the session. Trial types
 for the task are as follows. All mice 1: Free reward trial (reward at
 1001) 23: Light stimulation (20 ms pulse stimulation at 1001) 25: Light
 stimulation (50 ms pulse stimulation at 1001) 26: Light stimulation (250
 ms pulse stimulation at 1001)   **Files start from “OptoCtrl_Glu_”** Mouse
 ID: 591, 607, and 611 Glutamate sensor signal data during free light
 stimulation (single 20, 50, or 250 ms pulse) and free reward trials in the
 mice expressing tdTomato in glutamate input to the VTA are stored in these
 files. “dFoverFGCaMP2” is the z-scored glutamate signal for each trial in
 a single session. -1 sec to 8 sec from water or airpuff onset.
 “lickmatrix” is the lick event stamp (1: lick, 0: no lick) for the same
 time window as photometry data. “TrialType” is the list of trial type
 order in the session. Trial types for the task are as follows. All mice 1:
 Free reward trial (reward at 1001) 23: Light stimulation (20 ms pulse
 stimulation at 1001) 25: Light stimulation (50 ms pulse stimulation at
 1001) 26: Light stimulation (250 ms pulse stimulation at 1001)   **Files
 start from “OptoChr_DA_”** Mouse ID: 589, 692, 693, and 608 Dopamine
 sensor signal data during free light stimulation (single 20, 50, or 250 ms
 pulse) and free reward trials in the mice expressing ChrimsonR in
 glutamate input to the VTA are stored in these files “dFoverFGCaMP2” is
 the z-scored dopamine sensor signal for each trial in a single session. -1
 sec to 8 sec from water or airpuff onset. “lickmatrix” is the lick event
 stamp (1: lick, 0: no lick) for the same time window as photometry data.
 “TrialType” is the list of trial type order in the session. Trial types
 for the task are as follows. All mice 1: Free reward trial (reward at
 1001) 23: Light stimulation (20 ms pulse stimulation at 1001) 25: Light
 stimulation (50 ms pulse stimulation at 1001) 26: Light stimulation (250
 ms pulse stimulation at 1001)   **Files start from “OptoCtrl_DA_”** Mouse
 ID: 591, 607, and 611 Dopamine sensor signals from VTA neurons during free
 light stimulation (single 20, 50, or 250 ms pulse) and free reward trials
 in the mice expressing tdTomato in glutamate input to the VTA are stored
 in these files. “dFoverFGCaMP2” is the z-scored dopamine sensor signal for
 each trial in a single session. -1 sec to 8 sec from water or airpuff
 onset. “lickmatrix” is the lick event stamp (1: lick, 0: no lick) for the
 same time window as photometry data. “TrialType” is the list of trial type
 order in the session. Trial types for the task are as follows. All mice 1:
 Free reward trial (reward at 1001) 23: Light stimulation (20 ms pulse
 stimulation at 1001) 25: Light stimulation (50 ms pulse stimulation at
 1001) 26: Light stimulation (250 ms pulse stimulation at 1001)