Abstract: This dataset includes the source data related to the submission of the article “Sex differences in fear memory consolidation via Tac2 signaling in mice” to Nature Communications

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This documentation file was generated on 2021-03-02 by Antonio Florido and Raul Andero.

GENERAL INFORMATION ---

1. Title of Dataset: Effects of Tac2 pathway blockade on fear memory consolidation, circulating steroids expression, and mRNA and protein expression in the Central Amygdala.

2. Authorship:

Name: Raül Andero Galí

Institution: Institut de Neurociències (UAB) Email: Raul.Andero@uab.cat

ORCID: 0000-0003-3641-8903 Name: Antonio Florido

Institution: Institut de Neurociències (UAB) Email: AntonioLuis.Florido@uab.cat

ORCID: 0000-0001-6184-0706

DESCRIPTION ---

1. Dataset language: English

2. Abstract:

This dataset includes the source data related to the submission of the article “Sex differences in fear memory consolidation via Tac2

signaling in mice” to Nature Communications. It includes fear memory consolidation affections after the administration of a Neurokinin 3 Receptor antagonist including behavioral and molecular data related to sex-steroids and amygdalar mRNA and protein expression.

3. Keywords:

Sex-differences, Tac2 pathway, Amygdala.

4. Date of data collection (single date or date range):

2017-03-01 – 2021-02-01

5. Date of dataset publication:

2020-08-12

6. Funding sources:

Funding agency: Brain & Behavior Research Foundation Project number: NARSAD Young Investigator Grant #22434 Funding agency: Ministry of Science (Spain)

Project number: Ramón y Cajal program RYC2014-15784 Funding agency: Ministry of Economy (Spain)

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Project number: SAF2016-76565-R Funding agency: Ministry of Treasury Project number: FEDER funds

Funding agency: Federal Ministry of Education and Research Project number: NEURON-ERANET JTC 2019

Funding agency: Instituto de Salud Carlos III Project number: AC19/00077

Funding agency: Ministry of Science and Innovation (Spain) Project number: SAF2016-80027-R

Funding agency: Ministry of Science and Innovation (Spain) Project number: PID2019-106615RB-100

Funding agency: Instituto de Salud Carlos III Project number: CB06/05/0042

Funding agency: Ministry of Science and Innovation (Spain) Project number: SAF2017-83430

Funding agency: Federal Ministry of Education and Research Project number: NEURON-ERANET JTC 2019

Funding agency: ICREA

Project number: ICREA Academia Award (Generalitat de Catalunya, 2015-2019)

Funding agency: Generalitat de Catalunya Project number: 2018 FI_B00030

Funding agency: Generalitat de Catalunya Project number: 2017 FI_B00326

Funding agency: Ministry of Economy (Spain) Project number: FPI-2017

Funding agency: Ministry of Science, Innovation and Universities (Spain)

Project number: FJC2018-037958-I

Funding agency: Ministry of Science, Education, Culture and Sport (Spain)

Project number: FPU16/05416

Funding agency: Spanish National Health System Project number: CPII16/00027

7. Geographic location/s of data collection:

Barcelona, Spain.

ACCESS INFORMATION ---

1. Creative Commons License of the dataset:

Creative Commons License

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2. Dataset DOI:

3. Related publication:

Antonio Florido, Eric Velasco, Carlos Soto, Alex Gómez, Roser Nadal, Oscar Pozo, Carlos Saura, Raül Andero. Opposite-sex effects of the Tac2 pathway blockade in fear memory consolidation. Research Square.

DOI: 10.21203/rs.3.rs-38533/v1 [Preprint]

The article has been reviewed in Nature Communications.

VERSIONING AND PROVENANCE --- 1. Last modification date:

2021-02-01

2. Was data derived from another source?:

No

3. Additional related data collected that was not included in the current data package:

METHODOLOGICAL INFORMATION ---

1. Description of methods used for collection-generation of data:

Adult and young male and female mice underwent a fear conditioning procedure (five pairings of 30s, 60KHz tone coterminating with a footshock) and received osanetant (Nk3R antagonist, 5mg/Kg,

intraperitoneally, ip) 30 minutes after. Animals were tested for fear memory with the presentation of 15 tones in a different context 24 hours after the administration of the vehicle or the drug (sheets 1, 2, and 3 of the document).

Adult and young male and female mice brains were sliced and stained with an anti-Nk3R antibody for determination of Nk3+ positive cells in the Central Amygdala using Fiji for Windows.

Adult male and female mice’s blood was collected 30 minutes and 330 minutes after the treatments and steroid determination was performed using Mass Spectrometry and ELISA (Testosterone, Corticosterone, Progesterone, Deoxycorticosterone, Dehydrocorticosterone, and Estradiol; sheets 4 and 5 of the document).

Female mice were implanted with a bilateral cannula aiming at the amygdala and received intra amygdalar osanetant after a fear

conditioning task to evaluate memory alteration 24 hours after the injection (sheet 6 of the document).

Male and naturally cycling female Tac2-Cre mice were inoculated with a Cre-dependent Adeno-Associated Virus for the expression of the

inhibitory DREADD hMD4i in Tac2 cells in the Central Amygdala. These neurons were silenced through an ip injection of CNO (1mg/Kg, ip) 30 minutes after fear conditioning and memory was tested 24 hours later (sheet 7 of the document). Tac2 neurons in the Central Amygdala were counted using Fiji for Windows (sheet 8 of the document).

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Amygdala lysates of male and female mice that received osanetant 30 minutes after fear conditioning were analyzed for protein expression of total CREB, phospho-CREB, Total Akt, phospho-Akt, Total mTOR,

phosphor-mTOR, Total GSK3, phospho-GSK3, Total -Catenin and phospho-

-Catenin (sheet 9 of the document).

Male mice were bilaterally cannulated in the Central Amygdala and, after undergoing a fear conditioning task, all received osanetant ip, and also received an intra-amygdalar injection of vehicle, Akt

activator, or Androgen Receptor agonist. Fear memory was tested 24 hours later.

Female mice were bilaterally cannulated in the Central Amygdala and, after a fear conditioning task, all received osanetant ip, and also received an intra-amygdala injection of vehicle, anti-Akt or Estrogen Receptors antagonist (Sheet 10 of the document).

Male and female mice were evaluated during fear conditioning and fear expression tests for darting behavior using recordings of the

sessions. Darting behavior was manually counted by an expert (sheet 11 of the document).

Male and naturally cycling female mice received 1mg/Kg and 10mg/Kg doses of osanetant ip 30 minutes after fear conditioning to evaluate the effect of these doses 24 hours later during fear expression (sheet 12 of the document).

Male and naturally cycling female mice underwent the confrontation tube test in which, after housing animals in pairs for 5 weeks, each pair confront inside a methacrylate tube for 6 sessions (8

trials/session) to assess the dominant (which one stays inside of the tube) versus the submissive mice of the pair (sheet 13 of the

document).

Dominant and submissive male and female mice received osanetant 5mg/Kg after a fear conditioning task and were evaluated for fear memory recall 24 hours later (sheet 14 of the document). Blood of these animals was collected by tail-nick in basal conditions, before

habituation to the confrontation tube, before the confrontation tube test, before habituation to the fear conditioning procedure, before fear conditioning, and before fear expression test; and corticosterone values were measured by Mass Spectrometry (sheet 15 of the document).

Corticosterone and Testosterone were measured 330 minutes in amygdala micropunches of male mice using Mass Spectrometry (sheet 16 of the document).

Estradiol was assessed by Mass Spectrometry using serum from females that were decapitated under basal conditions (sheet 17 of the

document).

Brain slices of male and female mice were stained with antibodies for Nk3R, Estrogen receptor alpha and beta, GAD65, CaMKIIalpha and vGLUT2.

Colocalization with Nk3R was performed using Fiji for windows (sheet 18 of the document).

Male and naturally cycling female mice received osanetant after a fear conditioning task and, 30 minutes after, both amygdalae were extracted for qPCR of G-Protein Coupled Receptor related genes (sheet 19 of the document).

All procedures’ description is explained in detail in the preprint version of the submitted article:

Antonio Florido, Eric Velasco, Carlos Soto, Alex Gómez, Roser Nadal, Oscar Pozo, Carlos Saura, Raül Andero. Opposite-sex effects of the Tac2 pathway blockade in fear memory consolidation. Research Square.

DOI: 10.21203/rs.3.rs-38533/v1 [Preprint]

2. Methods for processing the data:

Freezing data was measured by automated software from Harvard

Apparatus (Panlab). Darting counts and confrontation tube test exits

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were analyzed by an experimented researcher. Amygdala micrographs were obtained using confocal microscopes and later analyzed using Fiji for windows expression or colocalization plugins. Sex steroids

determination, protein expression, and mRNA expression were determined by the controlling software for each dataset.

3. Instrument- or software-specific information needed to interpret the data:

Freezing v1.3.04 (Panlab-Harvard, Barcelona, Spain): used for

controlled delivery of tones and shocks and registration of freezing behavior through a high sensitivity weight transducer system. Target Lynx module of the MassLynx v4.1 (Water Associates, MA, USA): used for quantification by external calibration approach of plasma and

micropunch steroids. Fiji v1.52.n for Windows was used for confocal image processing. LungJ and Just Another Colocalization Plugin (JACoP) plugins were utilized to create and apply masks, and quantification of Mander's coefficient respectively. 7500FAST v2.0.6 (Thermo Fisher, Barcelona, Spain) software was employed as a controller for the thermocycler 7500FAST for quantification of mRNA in amygdala micropunches. Ingenuity Pathway Analysis (IPA) #44691306 (Qiagen, Germany) software was used to analyze canonical pathways containing identified significantly altered gene expression. ImageLab v5.2.1 software (Bio-Rad, Madrid Spain) was used for the quantification of Western Blots. Zeiss LSM700 Confocal microscope was controlled with ZEN v2010 software (Zeiss, Madrid, Spain). Leica TCS SP5 Confocal microscope was controlled by LAS X v2.7.3.9729 software (Leica, Madrid, Spain).

4. Instruments, calibration and standards information:

Freezing procedures were carried out using the StartFear system by Panlab-Harvard Apparatus. ABI 7500FAST thermocycler (Thermo Fisher) was used for qPCR arrays. Zeiss LSM700 Confocal microscope and Leica TCS SP5 Confocal microscope were used to obtain amygdala micrographs.

FILE OVERVIEW ---

1. Explain the file naming convention, if applicable:

2. File List:

Filename: Source_Data_Nat_Comm

Short description: this file contains the data reported in our submission to Nature Communications entitled “Sex differences in fear memory consolidation via Tac2 pathway in mice”.

3. Relationship between files:

4. File format: .xls

TABULAR DATA-SPECIFIC INFORMATION ---

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Sheet 1

1. File name: Males_osanetant

2. Number of rows and columns: 28 rows, 24 columns.

3. Variable list:

Variable name: ID

Description: Identification of the animal Measurement units or value labels: number Variable name: Treatment

Description: Treatment that the animal received after fear acquisition

Measurement units or value labels: vehicle or osanetant Variable name: Habituation

Description: Time spent freezed during 300 seconds prior to CS presentations

Measurement units or value labels: seconds Variable name: CS1

Description: Time spent freezed during the first CS presentation

Description: Time spent freezed during the second CS presentation

Description: Time spent freezed during the third CS presentation

Description: Time spent freezed during the fourth CS presentation

Description: Time spent freezed during the fifth CS presentation

Description: Time spent freezed during the sixth CS presentation

Variable name: CS7

Description: Time spent freezed during the seventh CS presentation

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Description: Time spent freezed during the eight CS presentation

Variable name: CS9

Description: Time spent freezed during the ninth CS presentation

Description: Time spent freezed during the tenth CS presentation

Description: Time spent freezed during the eleventh CS presentation

Description: Time spent freezed during the twelfth CS presentation

Description: Time spent freezed during the thirteenth CS presentation

Description: Time spent freezed during the fourteenth CS presentation

Description: Time spent freezed during the fifteenth CS presentation

Measurement units or value labels: seconds

4. Missing data codes:

Code or symbol: blank cell

Definition: loss value during measurement

Sheet 2

1. File name: Females_osanetant

3. Variable list:

Variable name: ID

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Measurement units or value labels: vehicle or osanetant Variable name: Estrous

Description: Estrous cycle stage during fear conditioning

Measurement units or value labels: proestrus, estrus, metestrus or diestrus

Variable name: Habituation

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Variable name: CS11

Description: Time spent freezed during the twelveth CS presentation

Definition: loss value during measurement Sheet 3

1. File name: Pups_osanetant

3. Variable list:

Variable name: ID

Description: Identification of the animal Measurement units or value labels: number Variable name: Sex

Description: Sex of the animal

Measurement units or value labels: Male or female Variable name: Treatment

Measurement units or value labels: vehicle or osanetant Variable name: Estrous

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Variable name: CS5

Variable name: CS9

Variable name: CS11

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Variable name: CS13

Sheet 4

1. File name: Nk3Rneurons

3. Variable list:

Variable name: ID

Measurement units or value labels: Male or female Variable name: Age

Description: Age of the animal

Measurement units or value labels: 8 weeks or P28 Variable name: Sex

Measurement units or value labels: Male or female Variable name: Estrous

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Variable name: Nk3R+

Description: number of Nk3R+ neurons

Measurement units or value labels: numbers Variable name: Nk3R+/mm²

Description: number of Nk3R+ neurons per mm²

Measurement units or value labels: positive cells/mm² 4. Missing data codes:

Sheet 5

1. File name: Steroids

3. Variable list:

Variable name: ID

Description: Identification of the animal Measurement units or value labels: number Variable name: Sex_Estrous

Description: Sex of the animal or estrous in females

Measurement units or value labels: Male or proestrus females or metestrus females

Variable name: Treatment

Description: Treatment received after fear acquisition Measurement units or value labels: vehicle or osanetant Variable name: Time point

Description: minutes after receiving the treatment until blood collection

Measurement units or value labels: minutes Variable name: Testosterone

Description: Serum testosterone

Measurement units or value labels: ng/ml Variable name: Corticosterone

Description: Serum corticosterone

Measurement units or value labels: ng/ml Variable name: Dehydrocorticosterone Description: Serum dehydrocorticosterone

Measurement units or value labels: ng/ml Variable name: Deoxycorticosterone

Description: Serum deoxycorticosterone Measurement units or value labels: ng/ml Variable name: Progesterone

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Description: Serum Progesterone

Measurement units or value labels: ng/ml

Code or symbol: Undetermined

1. File name: Estradiol

3. Variable list:

Variable name: ID

Measurement units or value labels: Female Variable name: Estrous

Description: Estrous cycle during conditioning

Measurement units or value labels: Proestrus, estrus, metestrus or diestrus

Description: Treatment received after fear acquisition Measurement units or value labels: vehicle or osanetant Variable name: Time point

Description: minutes after receiving the treatment until blood collection

Measurement units or value labels: minutes Variable name: Estradiol

Description: Serum estradiol

Measurement units or value labels: g/ml

1. File name: Proestrus_intraCeA

3. Variable list:

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Variable name: ID

Measurement units or value labels: vehicle or osanetant Variable name: Habituation

Variable name: CS5

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Description: Time spent freezed during the nineth CS presentation

Description: Time spent freezed during the twelveth CS presentation

Variable name: CS13

1. File name: DREADDs

3. Variable list:

Variable name: ID

Measurement units or value labels: Male or female

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Variable name: Estrous

Variable name: Genotype

Description: Genotype of the animal regarding Tac2-Cre expression

Measurement units or value labels: Wild-type or Heterozygous Variable name: Habituation

Variable name: CS5

Variable name: CS9

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Variable name: CS10

Variable name: CS11

Variable name: CS13

1. File name: CeM-mCherryNeurons

3. Variable list:

Variable name: ID

Description: Identification of the animal Measurement units or value labels: number Variable name: Genotype

Description: Genotype of the animal regarding Tac2-Cre expression

Measurement units or value labels: Wild-type or Heterozygous Variable name: mCherry+ neurons

Description: neurons expressing mCherry Measurement units or value labels: number Variable name: mCherry+/mm²

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Description: neurons expressing mCherry per mm² Measurement units or value labels: number

1. File name: WesternBlots

3. Variable list:

Variable name: ID

Description: Treatment received after fear acquisition

Measurement units or value labels: vehicle, osanetant or basal (without fear procedure)

Variable name: Sex

Measurement units or value labels: Male or Female Variable name: Total CREB

Description: Total CREB expressed in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: CREB(S113)/Total CREB

Description: Ratio CREB(S113)/Total CREB expressed in the Central Amygdala

Measurement units or value labels: Fold changes Variable name: Total Akt

Description: Total Akt expressed in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Akt(S473)/Total

Description: Ratio Akt(S473)/Total Akt expressed in the Central Amygdala

Measurement units or value labels: Fold changes Variable name: Akt(T308)/Total

Description: Ratio Akt(T308)/Total Akt expressed in the Central Amygdala

Measurement units or value labels: Fold changes

Variable name: Total mTOR

Description: Total mTOR expressed in the Central Amygdala Measurement units or value labels: Fold changes

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Variable name: mTOR(S2448)/Total

Description: Ratio mTOR(S2448)/Total mTOR expressed in the Central Amygdala

Measurement units or value labels: Fold changes Variable name: Total GSK3

Description: Total GSK3 expressed in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: GSK3(S9)/Total

Description: Ratio mTOR(S2448)/Total mTOR expressed in the Central Amygdala

Measurement units or value labels: Fold changes Variable name: Total -Catenin

Description: Total -Catenin expressed in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: GSK3(S9)/Total

Description: Ratio -Catenin(Ser33/37, Thr41)/Total mTOR expressed in the Central Amygdala

Measurement units or value labels: Fold changes 4. Missing data codes:

1. File name: Cannulations

3. Variable list:

Variable name: ID

Description: Estrous cycle stage during fear conditioning Measurement units or value labels: proestrus

Variable name: ip

Description: treatment received intraperitoneally Measurement units or value labels: osanetant Variable name: intra-CeA

Description: treatment received intra Central Amygdala

Measurement units or value labels: vehicle, SC 79, CI-4AS-1, Akti-1/2, ICI 182,780

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Variable name: CS5

Variable name: CS7

Variable name: CS11

(21)

Variable name: CS13

Sheet 12

1. File name: Darting

3. Variable list:

Variable name: ID

Description: Estrous cycle stage during fear conditioning Measurement units or value labels: proestrus

Description: Darts count during 300 seconds prior to CS presentations

Measurement units or value labels: number Variable name: CS1

Description: Darts count during the first CS presentation Measurement units or value labels: number

Variable name: CS2

Description: Darts count during the second CS presentation Measurement units or value labels: number

Variable name: CS3

Description: Darts count during the third CS presentation

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Measurement units or value labels: number Variable name: CS4

Description: Darts count during the fourth CS presentation Measurement units or value labels: number

Variable name: CS5

Description: Darts count during the fifth CS presentation Measurement units or value labels: number

Variable name: CS6

Description: Darts count during the sixth CS presentation Measurement units or value labels: number

Variable name: CS7

Description: Darts count during the seventh CS presentation Measurement units or value labels: number

Variable name: CS8

Description: Darts count during the eight CS presentation Measurement units or value labels: number

Variable name: CS9

Description: Darts count during the ninth CS presentation Measurement units or value labels: number

Variable name: CS10

Description: Darts count during the tenth CS presentation Measurement units or value labels: number

Variable name: CS11

Description: Darts count during the eleventh CS presentation Measurement units or value labels: number

Variable name: CS12

Description: Darts count during the twelfth CS presentation Measurement units or value labels: number

Variable name: CS13

Description: Darts count during the thirteenth CS presentation Measurement units or value labels: number

Variable name: CS14

Description: Darts count during the fourteenth CS presentation Measurement units or value labels: number

Variable name: CS15

Description: Darts count during the fifteenth CS presentation Measurement units or value labels: number

1. File name: Dose_Response

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3. Variable list:

Variable name: ID

Description: Estrous cycle stage during fear conditioning Measurement units or value labels: proestrus or metestrus Variable name: Osanetant dose ip

Measurement units or value labels: vehicle, osanetant 1mg/Kg or osanetant 10mg/Kg

Variable name: CS7

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Variable name: CS9

Description: Time spent freezed during the nineth CS presentation

Sheet 14

1. File name: CTT

3. Variable list:

Variable name: Sex

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Measurement units or value labels: Male or female Variable name: Pair

Description: Identification of each pair of animals Measurement units or value labels: number

Variable name: Session

Description: Session of the CTT

Measurement units or value labels: Session 1, Session 2, Session 3, Session 4, Session 5, Session 6.

Variable name: Trial per session

Description: Number of trial in each session

Measurement units or value labels: vehicle, osanetant 1mg/Kg or osanetant 10mg/Kg

Variable name: Winner

Description: Winner mouse per trial

Measurement units or value labels: 1 or 2 Variable name: D

Description: Dominant mouse per session Measurement units or value labels: 1 or 2 Variable name: Dominant

Description: Overall dominant animal Measurement units or value labels: 1 or 2

Code or symbol: N

Sheet 15

1. File name: FC_CTT

3. Variable list:

Variable name: ID

Description: Estrous cycle stage during fear conditioning Measurement units or value labels: proestrus or metestrus Variable name: Hierarchy

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Description: Hierarchical position for each mouse

Measurement units or value labels: Dominant or submissive Variable name: Habituation

Variable name: CS7

Variable name: CS9

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Variable name: CS12

Sheet 16

1. File name: Corticosterone_CTT

3. Variable list:

Variable name: ID

Measurement units or value labels: Male or females Variable name: Hierarchy

Description: Hierarchical position for each animal

Measurement units or value labels: dominant or submissive Variable name: Before habituation to CTT

Description: Corticosterone in serum before habituation to CTT Measurement units or value labels: ng/ml

Variable name: Before CTT

Description: Corticosterone in serum before CTT Measurement units or value labels: ng/ml

Variable name: Before habituation to FA

Description: Corticosterone in serum before habituation to fear conditioning chambers

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Measurement units or value labels: ng/ml Variable name: Before FA

Description: Corticosterone in serum before fear conditioning Measurement units or value labels: ng/ml

Variable name: Before FE

Description: Corticosterone in serum before fear expression test

1. File name: Punches_steroids

3. Variable list:

Variable name: ID

Measurement units or value labels: Male Variable name: Treatment

Description: Treatment administered after fear conditioning Measurement vehicle or osanetant

Variable name: Testosterone

Description: Testosterone concentration in amygdala micropunches

Measurement units or value labels: ng/ml Variable name: Corticosterone

Description: Corticosterone concentration in amygdala micropunches

Code or symbol: Undetermined Definition: loss value during measurement Sheet 16

1. File name: Corticosterone_CTT

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3. Variable list:

Variable name: ID

Measurement units or value labels: Male or females Variable name: Hierarchy

Description: Hierarchical position for each animal

Measurement units or value labels: dominant or submissive Variable name: Before habituation to CTT

Description: Corticosterone in serum before habituation to CTT Measurement units or value labels: ng/ml

Variable name: Before CTT

Description: Corticosterone in serum before CTT Measurement units or value labels: ng/ml

Variable name: Before habituation to FA

Description: Corticosterone in serum before habituation to fear conditioning chambers

Measurement units or value labels: ng/ml Variable name: Before FA

Description: Corticosterone in serum before fear conditioning Measurement units or value labels: ng/ml

Variable name: Before FE

Description: Corticosterone in serum before fear expression test

Sheet 18

1. File name: Estradiol_Mass_Spec

3. Variable list:

Variable name: ID

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Measurement units or value labels: Female Variable name: Estrous

Description: Estrous cycle stage during blood collection Measurement units or value labels: Proestrus or metestrus Variable name: Treatment

Description: Treatment administered Measurement units or value labels: Basal Variable name: E2 concentration

Description: Concentration of Estradiol in serum samples Measurement units or value labels: g/ml

Code or symbol: Undetermined Definition: loss value during measurement

Sheet 19

1. File name: Colocalization

3. Variable list:

Variable name: Sex

Measurement units or value labels: Male or Female Variable name: Estrous

Description: Estrous cycle stage during brain collection Measurement units or value labels: Proestrus or metestrus Variable name: Nk3R with ER

Description: Colocalization of Nk3R with ER in the Central Amygdala

Measurement units or value labels: Percentage Variable name: Nk3R with ER

Description: Colocalization of Nk3R with ER in the Central Amygdala

Measurement units or value labels: Percentage Variable name: Nk3R with GAD65

Description: Colocalization of Nk3R with GAD65 in the Central Amygdala

Measurement units or value labels: Percentage Variable name: Nk3R with CaMKII

Description: Colocalization of Nk3R with CaMKII in the Central Amygdala

Measurement units or value labels: Percentage

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Variable name: Nk3R with vGLUT2

Description: Colocalization of Nk3R with vGLUT2 in the Central Amygdala

Measurement units or value labels: Percentage 4. Missing data codes:

Code or symbol: Undetermined Definition: loss value during measurement Sheet 20

1. File name: qPCR_array

3. Variable list:

Variable name: ID

Measurement units or value labels: Male or Female Variable name: Estrous

Description: Estrous cycle stage during brain collection Measurement units or value labels: Proestrus or metestrus Variable name: Treatment

Description: Treatment administered 30 after fear conditioning Measurement units or value labels: Vehicle or osanetant

Variable name: Adcy5

Description: mRNA expression of Adcy5 in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Adora2a

Description: mRNA expression of Adora2a in the Central Amygdala

Measurement units or value labels: Fold changes Variable name: Adrb1

Description: mRNA expression of Adrb1 in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Adrb2

Description: mRNA expression of Adrb2 in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Agt

Description: mRNA expression of Agt in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Agtr1a

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Description: mRNA expression of Agtr1a in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Agtr1b

Description: mRNA expression of Agtr1b in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Agtr2

Description: mRNA expression of Agtr2 in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Agtrap

Description: mRNA expression of Agtrap in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Akt1

Description: mRNA expression of Akt1 in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Arrb1

Description: mRNA expression of Arrb1 in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Arrb2

Description: mRNA expression of Arrb2 in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Bai1

Description: mRNA expression of Bai1 in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Bcl2

Description: mRNA expression of Bcl2 in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Bcl2l1

Description: mRNA expression of Bcl2l1 in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Calcr

Description: mRNA expression of Calcr in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Calcrl

Description: mRNA expression of Calcrl in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Casr

Description: mRNA expression of Casr in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Ccl2

Description: mRNA expression of Ccl2 in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Ccl4

Description: mRNA expression of Ccl4 in the Central Amygdala Measurement units or value labels: Fold changes

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Variable name: Ccnd1

Description: mRNA expression of Ccnd1 in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Ccne1

Description: mRNA expression of Ccne1 in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Ccne2

Description: mRNA expression of Ccne2 in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Cdkn1a

Description: mRNA expression of Cdkn1a in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Cdkn1b

Description: mRNA expression of Cdkn1b in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Cflar

Description: mRNA expression of Cflar in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Col1a1

Description: mRNA expression of Col1a1 in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Crhr1

Description: mRNA expression of Crhr1 in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Crhr2

Description: mRNA expression of Crhr2 in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Ctgf

Description: mRNA expression of Ctgf in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Cyp19a1

Description: mRNA expression of Cyp19a1 in the Central Amygdala

Measurement units or value labels: Fold changes Variable name: Drd1a

Description: mRNA expression of Drd1a in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Drd2

Description: mRNA expression of Drd2 in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Dusp14

Description: mRNA expression of Dusp14 in the Central Amygdala Measurement units or value labels: Fold changes

(34)

Variable name: Edn1

Description: mRNA expression of Edn1 in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Egr1

Description: mRNA expression of Egr1 in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Elk4

Description: mRNA expression of Elk4 in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Fgf2

Description: mRNA expression of Fgf2 in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Fos

Description: mRNA expression of Fos in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Galr2

Description: mRNA expression of Galr2 in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Gcgr

Description: mRNA expression of Gcgr in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Gnaq

Description: mRNA expression of Gnaq in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Gnas

Description: mRNA expression of Gnas in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Grm1

Description: mRNA expression of Grm1 in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Grm2

Variable name: Grm4

Variable name: Grm5

Variable name: Grm7

Variable name: Icam1

(35)

Description: mRNA expression of Icam1 in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Il1b

Description: mRNA expression of Il1b in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Il1r1

Description: mRNA expression of Il1r1 in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Il1r2

Description: mRNA expression of Il1r2 in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Il2

Description: mRNA expression of Il2 in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Jun

Description: mRNA expression of Jun in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Junb

Description: mRNA expression of Junb in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Kcnh8

Description: mRNA expression of Kcnh8 in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Lhcgr

Description: mRNA expression of Lhcgr in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Lpar1

Description: mRNA expression of Lpar1 in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Lpar2

Description: mRNA expression of Lpar2 in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Max

Description: mRNA expression of Max in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Mmp9

Description: mRNA expression of Mmp9 in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Myc

Description: mRNA expression of Myc in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Nos2

Description: mRNA expression of Nos2 in the Central Amygdala Measurement units or value labels: Fold changes

(36)

Variable name: Oprd1

Description: mRNA expression of Oprd1 in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Oprk1

Description: mRNA expression of Oprk1 in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Pdpk1

Description: mRNA expression of Pdpk1 in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Pik3cg

Description: mRNA expression of Pik3cg in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Prkca

Description: mRNA expression of Prkca in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Ptgdr

Description: mRNA expression of Ptgdr in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Ptgs2

Description: mRNA expression of Ptgs2 in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Rgs2

Description: mRNA expression of Rgs2 in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Rho

Description: mRNA expression of Rho in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: S1pr1

Description: mRNA expression of S1pr1 in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Sctr

Description: mRNA expression of Sctr in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Serpine1

Description: mRNA expression of Serpine1 in the Central Amygdala

Measurement units or value labels: Fold changes Variable name: Socs1

(37)

Description: mRNA expression of Socs1 in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Tnf

Description: mRNA expression of Tnf in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Tshr

Description: mRNA expression of Tshr in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Upc1

Description: mRNA expression of Upc1 in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Vcam1

Description: mRNA expression of Vcam1 in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Vegfa

Description: mRNA expression of Vegfa in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Ywhaz

Description: mRNA expression of Ywhaz in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Actb

Description: mRNA expression of Actb in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: B2m

Description: mRNA expression of B2m in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Gapdh

Description: mRNA expression of Gapdh in the Central Amygdala Measurement units or value labels: Fold changes

Variable name: Gusb (HK gene)

Description: mRNA expression of Gusb (HK gene)in the Central Amygdala

Measurement units or value labels: Fold changes Variable name: HSP90ab1

Description: mRNA expression of HSP90ab1in the Central Amygdala

Measurement units or value labels: Fold changes

Code or symbol: Undetermined Definition: loss value during measurement

(38)