Data from: Cilia-driven epithelial folding and unfolding in an early-diverging animal
Data files
Nov 29, 2025 version files 226.51 GB
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animal_3D_curvature_maps.zip
32.58 MB
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animal_folding_on_glass_capillaries.zip
4.17 GB
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animal_folding_snapshots_-_light_sheet.zip
14.96 GB
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animal_folding_without_substrate.zip
13.64 GB
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animal_unfolding_trials_-_LiCl_and_CFSW.zip
6.14 GB
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animal_unfolding_trials_-_light_sheet.zip
39.69 GB
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animal_unfolding_trials_-_size.zip
27.14 GB
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animal_unfolding_trials_-_unfolding_time_distribution.zip
119.87 GB
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cellular_perspective_in_folds_-_confocal.zip
623.95 MB
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README.md
5.91 KB
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silicone_rubber_sheet_unfolding.zip
223.27 MB
Abstract
Multicellular organisms utilize epithelial folding to achieve remarkable three-dimensional forms. During embryonic development, stereotypical epithelial folds emerge from underlying active cellular and molecular processes including cell shape change and differential cell growth. However, the origin of epithelial folding in early animals and how folding may be harnessed in synthetic systems remain open questions. Here we discover a novel modality of behavior-induced epithelial folding and unfolding arising from cilia-substrate adhesion and ciliary walking in the basal animal Trichoplax adhaerens (phylum Placozoa). We show that T. adhaerens is capable of exhibiting dynamic non-stereotyped folding states, providing a novel 3D perspective to an organism previously only characterized in its 2D state. We correlate these folding states to local substrate geometry, revealing that the animal conforms to available substrate surface area, promoting the maintenance of a folded state. Using 4D fluorescence light sheet microscopy, we characterize fold geometry, curvature evolution during unfolding, and the non-stereotypy of unfolding behavior. Through repeated unfolding trials, we reveal the robustness and timescales associated with unfolding behavior and employ scaling analysis and toy model simulations to establish how collective ciliary activity can robustly drive unfolding. In this way, despite lacking any folding-unfolding "pathway," transitions between folding and unfolding states emerge as a function of the animal’s environment and motility. Our work reveals a remarkable behavior exhibited by a brainless, nerveless animal, and demonstrates the capacity for 3D-2D transitions in folding epithelial sheets using ciliary activity.
Dataset DOI: 10.5061/dryad.xpnvx0kvd
Description of the data and file structure
The following datasets were collected in support of the study of cilia-driven folding and unfolding behavior exhibited by the early-diverging animal Trichoplax adhaerens. The headings below correspond to zip files included in this dataset. Animal numbering (e.g., "animal1") is not preserved throughout the dataset, and only serves to organize data within a given section.
1. animal folding on glass capillaries
This section contains data from experiments testing T. adhaerens' folding state as a function of substrate geometry. Animals were allowed to attach to the surface of a glass capillary of either 1mm or 0.17mm outer diameter, suspended in a dish of artificial sea water.
- The directory structure is
[capillary diameter] / [unfolding trial] animal_area_on_capillary.py: Python script to extract and plot the maximum diameter of the animal contour as it is in contact with the capillary.
2. animal folding without substrate
This section contains data from experiments testing T. adhaerens' folding state in the absence of a rigid substrate. Animals were imaged in artificial sea water suspended in an annular imaging chamber.
- The directory structure is
[animal] / [image dir], where the image_dir contains a maximum of 1000 images. tplax_area_no_substrate.py: Python script to extract and plot the maximum diameter of the animal contour as it is suspended in the vertical tracking microscope.
3. animal unfolding trials - size
This section contains data from experiments testing T. adhaerens' unfolding trajectory as a function of animal size.
- The directory structure is
[animal] / [unfolding trial number]. animal_size_unfolding_times.csv: Table of unfolding times in provided directories.animal_size_unfolding_time_regression.py: Python script to compute unfolding time statistics as a function of animal size, and plot animal size vs. unfolding time statistic with linear regression of the same.animal_size_unfolding_times.csvprovides the input data to this script.animal_size_unfolding_time_violinplot.py: Python script to visualize the distribution of unfolding times sorted by animal identity.animal_size_unfolding_times.csvprovides the input data to this script.
4. cellular perspective in folds - confocal
This section contains data from confocal imaging of fixed T. adhaerens' in a folded state.
fixed fold hoechst and membrane stain.czi: raw 2-channel imaging file from confocal microscopy session. Channels show membrane stain (red) and nuclear stain (green).fixed fold hoechst and membrane stain.ims: imaris file with oblique slicer to visualize the fold profile. Same data as infixed fold hoechst and membrane stain.czi.
5. animal folding snapshots - light sheet
This section contains data from light sheet imaging of live and fixed T. adhaerens' in a folded state.
- The directory structure is
[animal] / [data files] / [raw data] - Snapshot 6 is of a fixed animal while all other snapshots are of live animals.
6. silicone rubber sheet unfolding
This section contains data from unfolding time lapses of silicone rubber sheets on glass surfaces coated with silicone oil.
- This directory contains an image directory (labeled 1-10) and a corresponding results file (labeled
[n]_Results.csv). The results files contain pixel area values for imaging data.
rubber_sheet_unfolding.csv: Python script to visualize the contact area over time for rubber sheet unfolding time lapses. Plots data in results files in an overlay plot.
7. animal unfolding trials - light sheet
This section contains data from unfolding time lapses of fluorescently labeled live T. adhaerens captured via light sheet microscopy.
- This directory structure is
[animal id] / [time lapse part]
8. animal 3D curvature maps
This section contains local curvature maps of light sheet data, obtained using MeshLab.
9. animal unfolding trials - LiCl and CFSW
This section contains unfolding trials for animals treated in a sequence of artificial sea water, either calcium-free sea water (CFSW) or 200 mM lithium chloride, and artificial sea water again.
./CFSWtrials: contains calcium-free sea water unfolding trials../CFSWtrials/Raw imagescontains the image sequence for each unfolding trial.Ignore_[filename]files contain any frames to ignore due to segmentation issues.Results_[filename]files contain contact area data in pixels../LiCl trials: contains 200 mM lithium chloride in sea water unfolding trials../LiCl trials/Raw imagescontains the image sequence for each unfolding trial.Ignore_[filename]files contain any frames to ignore due to segmentation issues.Results_[filename]files contain contact area data in pixels.licl_cfsw_unfolding.py: Python script to plot contact area vs. time for all unfolding trials in this section, organized by treatment type and whether the trial is pre-treatment, treatment, or post-treatment.
10. animal unfolding trials -unfolding time distribution
This section contains unfolding time lapses used to obtain unfolding times across a distribution of animals.
./additional unfolding timelapses/: contains 45 additional unfolding time lapses which, together with the data included in (3) above, are used to obtain an unfolding time distribution.animal_size_unfolding_times_all.csv: Unfolding time data from 163 unfolding time lapses, annotated by animal identity, size, and trial.animal unfolding times distribution.py: Python script to plot the distribution of unfolding times inanimal_size_unfolding_times_all.csv.
