DOI: 10.5061/dryad.rv15dv4fn
title: Chemo- and optogenetic activation of hypothalamic Foxb1-expressing neurons and their terminal endings in the rostral-dorsolateral PAG leads to tachypnea, bradycardia, and immobility

description: Foxb1-expressing neurons occur in the dorsal premammillary nucleus (PMd)
 and further rostrally in the parvafox nucleus, a longitudinal cluster of
 neurons in the lateral hypothalamus of rodents. The descending projection
 of these Foxb1+ neurons end in the dorsolateral part of the periaqueductal
 gray (dlPAG). The functional role of the Foxb1+ neuronal subpopulation in
 the PMd and the parvafox nucleus remains elusive. In this study, the
 activity of the Foxb1+ neurons and of their terminal endings in the dlPAG
 was selectively altered by employing chemo- and optogenetic tools. Our
 results show that in whole-body barometric plethysmography,
 hM3Dq-mediated, global Foxb1+ neuron excitation activates respiration.
 Time-resolved optogenetic gain-of-function manipulation of the terminal
 endings of Foxb1+ neurons in the rostral third of the dlPAG leads to
 abrupt immobility and bradycardia. Chemogenetic activation of Foxb1+ cell
 bodies and ChR2-mediated excitation of their axonal endings in the dlPAG
 led to a phenotypical presentation congruent with a “freezing-like”
 situation during innate defensive behavior. The authors have no relevant
 financial or non-financial interests to disclose. This work was supported
 by the Swiss National Foundation grant 31003A_160325 to Marco R. Celio.
description: # Chemo- and optogenetic activation of hypothalamic Foxb1-expressing
 neurons and their terminal endings in the rostral-dorsolateral PAG leads
 to tachypnea, bradycardia, and immobility
 This data set contains raw
 data collected in whole-body plethysmography, open field, hot plate
 experiments with wild-type and transgenic mice. Optogenetic and
 chemogenetic actuators were used to activate *Foxb1*+ neurons of the
 parvafox nucleus and the dorsal premammillary nucleus. ## Description of
 the data and file structure The data set contains 3 directories, one for
 each type of experiments (i.e. WholeBodyPlethysmography, OpenField,
 HotPlate). 1\. **WholeBodyPlethysmography:** * Raw files containing data
 across all conditions and replicates recorded for a specific animal (e.g.
 AllCond_Animal...xlsx) * Saline injected condition 1-3 = BL1-BL3 (1-3
 indicates the replicate number; 3 replicates were performed on 3
 consecutive days) * Clozapine injected condition 1-3 = Clo1-Clo3 (1-3
 indicates the replicate number; 3 replicates were performed on 3
 consecutive days) * Clozapine-N-oxide injected condition 1-3 = Exp1-3 (1-3
 indicates the replicate number; 3 replicates were performed on 3
 consecutive days) * Column parameters are as follows: * Event = Column
 indicating when external signals (e.g. TTL pulses) were detected (not used
 in this data set) * Alarm = Column indication if a user-defined threshold
 was reached (not used in this data set) * Flow_Num = The number of the
 cycle * PIF = The Peak Inspiratory Flow is the highest value measured from
 the baseline.  The baseline is formed by connecting the start inspiration
 mark and the start expiration mark. * PEF = The Peak Expiratory Flow is
 the lowest value measured from the baseline * TV = Tidal Volume is the
 integral of the difference between the inspiration signal and the baseline
 expressed in flow units-seconds. The baseline is formed by connecting the
 start inspiration mark and the start expiration mark  * MV = Minute Volume
 is the product of TV and BPM * BPM = Breaths Per Minute is the reciprocal
 of the period, in seconds, associated with a breath, multiplied by 60 * IT
 = Inspiration Time is the time, in milliseconds, from the start of
 inspiration to the end of inspiration. The start of inspiration is marked
 at the point where the respiration signal crosses the baseline with a
 positive slope. The end of inspiration signal is marked at the point where
 the respiration signal crosses the baseline with a negative slope * ET =
 Expiration Time is the time, in milliseconds, from the end of inspiration
 to the end of expiration * TT = Total Time is the sum of inspiration and
 expiration times: IT + ET * AT = Apnea Time (not used) * Pause = Pause
 provides a timing comparison of early and late expiration * PEnh =
 Enhanced Pause is the product of the ration of PEF to PIF and Pause *
 TVadj = The adjusted Tidal Volume is calculated by multiplying the
 reported TV by an adjustment factor based on the chamber temperature,
 relative humidity and atmospheric pressure * MVadj = The Minute Volume adj
 is the product of TVadj and BPM * ApCt = Apnea count (not used) * IF50 =
 Inspiratory Flow at 50% Tidal Volume * EF50 = Expiratory Flow at 50% Tidal
 Volume * E/IF50 = The ratio of Expiratory Flow to Inspiratory Flow at 50%
 Tidal Volume * Pamb = ambient pressure * T_Num = temperature cycle number
 * T_Mean = temperature mean * T_RMax = relative temperature maximum *
 T_RMin = relative temperature minimum * T_Per = temperature period * T_BPM
 = temperature breaths per minute * T_Area = temperature area * R_Num =
 relative humidity cycle number * R_Mean = relative humidity mean * R_Max =
 relative humidity maximum * R_Min = relative humidity minimum * R_Period =
 relative humidity period * R_BPM = relative humidity breaths per minute *
 R_Area = relative humidity area * TVadjPerGram(uL) = TVadj normalized to
 body weight (in microliters) * MVadjPerGram(mL) = MVadj normalized to body
 weight (in mililiters) * Correction_factor(TVadj/Tv) = Drorbaugh &
 Fenn Correction factor applied to correct raw values for recording
 conditions * PIFadj = adjusted Peak Inspiratory Flow (see
 Correction_factor) * PEFadj = adjusted Peak Expiratory Flow (see
 Correction_factor) * Condition = Experimental condition (BL = Baseline,
 Clo = Clozapine, Exp = Clozapine-N-oxide) * Animal = Animal number (ID) *
 DREADD_receptor = Which chemogenetic actuator was expressed (hM3Dq, hM4Di,
 or none (DREADD_neg)) * "DREADD_Additional_Information.xlsx"
 helper file containing animal ID and information 2\. **OpenField:** *
 Standard raw tracking data obtained by Deeplabcut
 ( * The files
 containing the raw data end in
 "...DLC_resnet50_OpenFieldDec23shuffle1_600000_filtered.csv" *
 The section just prior to the above mentioned suffix contains the
 condition and replicate information (e.g. BL2 for baseline replicate 2) *
 Files with a different suffix are helper files necessary for the analysis
 and interpretation. * Tracked body parts and landmarks are labeled as: *
 ArenaTL/ArenaTR/ArenaBL/ArenaBR = Corners of the open field arena
 (top-left, top-right, bottom-left, bottom-right) * Body parts: Nose,EarR
 (base of right ear),EarL (base of left ear), Occiput, TailBase *
 HelperFile column parameters: * VideoName_AnimalID = Identifier of the
 video recording associated with a given animal * genotype = Genotype of
 the mouse (all Foxb1-Cre, see associated publication for details) * sex =
 f for female; m for male * DREADDreceptro = Which chemogenetic actuator
 was expressed (hM3Dq, hM4Di, or none (DREADD_neg)) * WBPsession =
 Whole-body plethysmography recordings were performed with these mice. The
 recordings were either started at 9:00am or 10:30am. This column indicates
 in which sessions the mice were recorded for whole-body plethysmography,
 but this information is not required for the OpenField analysis. *
 PerfusionNr = unique identifier given to each mouse after perfusion *
 PerfusionDate = date on which mouse was perfused 3\. **HotPlate:** *
 Scoring data for different features measured in the hot plate experiments.
 * 23-01-01_Opto_HotPlate_1stLickAttempt_or_correctEP * This file contains
 average durations (in seconds) until the mouse either displayed it's
 first attempt to lick its hind paw, actually licked its hind paw or jumped
 (whichever happened first). * Animal = Animal identifier * BL_avg =
 Baseline average (no optogenetic stimulation) * Opto_avg = Average of the
 stimulation condition (either activation of ChR2 or ArchT3.0; see
 HelperFile parameters) * 23-01-01_Opto_HotPlate_JumpOrCorrectLick * This
 file contains average durations (in seconds) until the mouse either licked
 its hind paw or jumped (whichever happened first). * Animal = Animal
 identifier * BL_avg = Baseline average (no optogenetic stimulation) *
 Opto_avg = Average of the stimulation condition (either activation of ChR2
 or ArchT3.0; see HelperFile parameters) *
 23-01-01_Opto_HotPlate_nShakesBeforeEP * This file contains the average
 number of hind limb shakes before the mouse either licked its hind paw or
 jumped (whichever happened first). * Animal = Animal identifier * BL_avg =
 Baseline average (no optogenetic stimulation) * Opto_avg = Average of the
 stimulation condition (either activation of ChR2 or ArchT3.0; see
 HelperFile parameters) * 23-01-01_Opto_HotPlate_TimeTo1stShake * This file
 contains average durations (in seconds) until the mouse performed its
 first hind limb shake. * Animal = Animal identifier * BL_avg = Baseline
 average (no optogenetic stimulation) * Opto_avg = Average of the
 stimulation condition (either activation of ChR2 or ArchT3.0; see
 HelperFile parameters) * HelperFile column parameters: * AnimalNr = Animal
 ID given at birth of the animal * genotype = Genotype of mouse (Foxb1-Cre
 or PV-Cre (see associated publication for more details)) * sex = f for
 female; m for male * type of opsin = optogenetic modulators expressed in
 the mouse (ChR2 = Channelrhodopsin-2; ArchT3.0 = Archaerhodopsin-3) *
 Perfusion# = unique identifier given to each mouse after perfusion *
 Perfusion date = date on which mouse was perfused * FiberLocGroup = Group
 allocation based on position of optic fiber tip (see publication for more
 details) ## Sharing/Access information Data was derived from the following
 sources: * Raw data for the scRNAseq reanalysis was obtained from
 *
 Accompanying code to analyze the deposited data will be made available
 upon publication of the study on the github repository of the lead author