Chemo- and optogenetic activation of hypothalamic Foxb1-expressing neurons and their terminal endings in the rostral-dorsolateral PAG leads to tachypnea, bradycardia, and immobility
Data files
Jan 31, 2024 version files 308.03 MB
Abstract
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.
README: Chemo- and optogenetic activation of hypothalamic Foxb1-expressing neurons and their terminal endings in the rostral-dorsolateral PAG leads to tachypnea, bradycardia, and immobility
https://doi.org/10.5061/dryad.rv15dv4fn
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 (http://www.mackenziemathislab.org/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)
- This file contains average durations (in seconds) until the mouse either licked its hind paw or jumped (whichever happened first).
- 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)
- This file contains the average number of hind limb shakes before the mouse either licked its hind paw or jumped (whichever happened first).
- 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)
- This file contains average durations (in seconds) until the mouse performed its first hind limb shake.
- 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 https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE146692
- Accompanying code to analyze the deposited data will be made available upon publication of the study on the github repository of the lead author https://github.com/rbcola/