DOI: 10.5061/dryad.1vhhmgr01
title: A single neuron in C. elegans orchestrates multiple motor outputs through parallel modes of transmission

description: Animals generate a wide range of highly coordinated motor outputs, which
 allows them to execute purposeful behaviors. Individual neurons in the
 circuits that generate behaviors have a remarkable capacity for
 flexibility, as they exhibit multiple axonal projections, transmitter
 systems, and modes of neural activity. How these multi-functional
 properties of neurons enable the generation of adaptive behaviors remains
 unknown. Here we show that the HSN neuron in C. elegans evokes multiple
 motor programs over different timescales to enable a suite of behavioral
 changes during egg-laying. Using HSN activity perturbations and in vivo
 calcium imaging, we show that HSN acutely increases egg-laying and
 locomotion while also biasing the animals towards low-speed dwelling
 behavior over minutes. The acute effects of HSN on egg-laying and
 high-speed locomotion are mediated by separate sets of HSN transmitters
 and different HSN axonal projections. The long-lasting effects on dwelling
 are mediated by HSN release of serotonin that is taken up and re-released
 by NSM, another serotonergic neuron class that directly evokes dwelling.
 Our results show how the multi-functional properties of a single neuron
 allow it to induce a coordinated suite of behaviors and also reveal that
 neurons can borrow serotonin from one another to control behavior.
description: GCaMP datasets were collected from freely-moving <em>C.
 elegans</em> animals, using widefield imaging of animals on agar
 slides, as described in the manuscript.
description: <strong>ImageJ tracking script:</strong>  Put the
 .txt file and the .ijm file in the same folder under the
 "macros" folder in ImageJ/Fiji installation. The .txt file
 includes settings that can be adjusted for the neuron of interest. the
 .ijm file is an ImageJ macro <strong>Neuron
 retracking script</strong>: MATLAB code that enables tracking of
 fluorescence intensity in the same neuron location in different channel.
 The inputs required are a txt file containing the neuron location (which
 is the output of the ImageJ tracking script), and Tiff files from the
 recordings. <strong>For HSN GCaMP
 dataset:</strong> there are 6 file folders: 1) gcamp raw 2) mscarlet
 raw 3) Ratiometric are in .mat files (MATLAB) ; 4) Eggevetns 5) GcampPeaks
 and 6) speed are in txt file format  <strong>For
 BAG GCaMP dataset:</strong> each folder contains a JPEG file to
 define fused agar boundary and a gcamp tracking file (a csv with:
 frame,xcoord,ycoored,intdens,intsub,bgmedian,maxint,area,x,y,sqintdens,sqintsub,sqarea,threshold,animal,redFlag,useTracking)
description: ##A single neuron in C. elegans orchestrates multiple motor outputs
 through parallel modes of transmission This dataset is from the study “A
 single neuron in C. elegans orchestrates multiple motor outputs through
 parallel modes of transmission” by YC Huang, J Luo, W Huang, CM Baker, MA
 Gomes, AB Byrne, and SW Flavell (2023). GCaMP datasets were collected from
 freely-moving animals placed on agar with seeded OP50 bacterial food. The
 HSN neuron calcium imaging aimed to investigate the activity of the HSN
 neuron related to the animals' locomotion and egg-laying behavior on
 food. The BAG neuron calcium imaging aimed to observe whether the BAG
 neuron responds to the osmotic upshift. ## Description of the data and
 file structure The HSN\_GCaMP\_data folder contains six subfolders that
 provide HSN activity and animals' behaviors from indicated
 recordings, labeled with the date of recording in the file name. 1. The
 EggEvents folder contains .txt files that show the frame(s) in which an
 animal laid an egg during that recording. Recordings that are in other
 folders but absent in this folder are recordings without egg-laying
 events. 2. The gcamp\_raw folder contains .mat (MATLAB) files with
 background-subtracted HSN GCaMP intensity from the recordings. 3. The
 GcampPeaks folder contains .txt files that show the frames in which the
 recordings have maximum HSN GCaMP peaks. 4. The mscarlet\_raw folder
 contains .mat (MATLAB) files with background-subtracted HSN mscarlet
 intensity from the recordings. As the mscarlet doesn't respond to
 neuron activity changes, it is used as a correction of imaging focus
 shift. 5. The Ratiometric folder contains .mat (MATLAB) files with
 gcamp\_raw normalized to mscarlet\_raw. 6. The Speed folder contains .txt
 files for the speed of the animals in the corresponding recordings. For
 BAG calcium imaging, animals were placed on agar fused with two different
 osmolarities. Under the BAG\_GCaMP\_data folder, each subfolder contains a
 TIF file showing the location of the boundary of two osmolarities and a
 .txt file that contains information on BAG neuron activity and the
 neuron's location in the corresponding frames. The variables used
 from .txt files are from column 1-5: Slice (frame), xc (x-coordinate), yc
 (y-coordinate), intdens (integrated density), and intsub
 (background-subtracted integrated density). • Relationship between files,
 if important: None • Additional related data collected that was not
 included in the current data package: None • Are there multiple versions
 of the dataset? No ## Code/Software ImageJ 2.3 (Fiji) The "imagej
 scripts for tracking" folder contains a .txt file and 2 .ijm files.
 These scripts are used in the publication for automatic single neuron
 tracking from recorded videos. Please refer to the linked publication for
 more details about the imaging setup to see if this script is useful for
 your recordings or if parameters in the codes need to be modified. To use
 these scripts, put all files under the "macros" folder in your
 ImageJ/Fiji installation. The .txt file contains settings that you can
 modify to adjust tracking for your neuron. The .ijm file is an ImageJ
 macro. MATLAB (2022a) "Neuron\_retracking" is a MATLAB code that
 allows you to use the .txt files that are outputs from the ImageJ scripts
 to retrack the pixel intensity from the same pixel location from different
 channels (e.g., GFP vs mCherry) of the same recording. The input files for
 this function are the .txt file output from the ImageJ scripts and
 individual TIF files from recordings (videos need to be exported as
 individual TIF files first). ## Sharing/Access information 1. Links to
 publications that cite or use the data: YC Huang, J Luo, W Huang, CM
 Baker, MA Gomes, AB Byrne and SW Flavell (2023). A single neuron in C.
 elegans orchestrates multiple motor outputs through parallel modes of
 transmission 2. Links to other publicly accessible locations of the data:
 None 3. Links/relationships to ancillary data sets: None 4. Was data
 derived from another source? No A. If yes, list source(s): NA 5.
 Recommended citation for this dataset: YC Huang, J Luo, W Huang, CM Baker,
 MA Gomes, AB Byrne and SW Flavell (2023). A single neuron in C. elegans
 orchestrates multiple motor outputs through parallel modes of transmission
 Dryad Digital Repository. doi:10.5061/dryad.1vhhmgr01