##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