Rapid neural DNA methylation responses to predation stress in Trinidadian guppies
Data files
Apr 28, 2025 version files 101.57 KB
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data.zip
50.59 KB
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README.md
5.51 KB
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scripts.zip
45.46 KB
Apr 28, 2025 version files 101.63 KB
-
data.zip
50.59 KB
-
README.md
5.58 KB
-
scripts.zip
45.46 KB
Abstract
DNA methylation (DNAm) is a well-studied epigenetic mechanism implicated in environmentally induced phenotypes and phenotypic plasticity. However, few studies investigate the time scale of DNAm shifts. Thus, it is uncertain whether DNAm can change on timescales relevant for rapid phenotypic shifts, such as during the expression of short-term behavioural plasticity. DNAm could be especially reactive in the brain, potentially increasing its relevance for behavioural plasticity. Most research investigating neural changes in methylation has been conducted in mammalian systems, on isolated individuals, and using stressors that are less ecologically relevant, reducing their generalizability to other natural systems. We exposed pairs of male and female Trinidadian guppies (Poecilia reticulata) to alarm cue, conspecific skin extract that reliably induces anti-predator behaviour, or a control cue. Whole genome bisulfite sequencing on whole brains at various time points following cue exposure (0.5h, 1h, 4h, 24h, and 72h) allowed us to uncover the timescale of neural DNAm responses. Males and females both showed rapid shifts in DNAm in as little as 0.5 hours. However, males and females differed in the time-course of their responses: both sexes showed a peak in the number of loci showing significant responses at 4 hours but males showed an additional peak at 72 hours. We suggest that this finding could be due to differing longer-term plastic responses between the sexes. This study shows that DNAm can be rapidly induced by an ecologically relevant stressor in fish and suggests that DNA methylation could be involved in short-term behavioural plasticity.
https://doi.org/10.5061/dryad.x0k6djhw6
Description of the data and file structure
Rapid neural DNA methylation responses to predation stress in Trinidadian guppies
DOI: http://doi.org/10.1111/mec.17774.
Experiment Summary:
Adult guppies were exposed to either alarm cue or control cue one time. Behaviour was measured for 5 minute before and after cue exposure. At set time points following cue exposure (0.5h, 1h, 4h, 24h, 72h), brains were dissected out for whole genome bisulfite sequencing.
File structure:
Raw and clean behavioural and tank info in data files (raw and clean folders) are provided. Sequencing data are stored on SRA. Analysis scripts are provided in separate folders for each analysis type (01_behavioural_analysis and 02_methylation_analysis). Within each folder, each script should be run in order of numbering.
All variables and abbreviations defined in metadata. Missing data = NA.
Files and variables
File: data.zip
Description: zip file containing all data
File: all_cue_data.csv
Description: Cleaned full behavioural responses dataset (10 minutes of behaviour before and after cue exposure)
Variables
- obs_id: observation ID
- sex: sex (female/male)
- length_obs_s: length of time of observation
- sig_s: length of time engaging in sigmoidal displays (seconds) - males only
- sig_nb: number of sigmoidal displays - males only
- chasing_s: length of time spent chasing the female (seconds) - males only
- feeding_s: length of time spent feeding (seconds)
- frozen_s: length of time spent frozen (seconds)
- Q1_s: length of time in quadrant 1 (seconds)
- Q2_s: length of time in quadrant 2 (seconds)
- Q3_s: length of time in quadrant 3 (seconds)
- Q4_s: length of time in quadrant 4 (seconds)
- Q5_s: length of time in quadrant 5 (seconds)
- Q6_s: length of time in quadrant 6 (seconds)
- dash_nb: Number of dashes
- bin: which time bin observation is a part of
- tank: tank ID number
- time: before or after cue exposre
- fish_ID: fish ID
- cue: which cue tank was exposed to (alarm or control)
File: clean_cue_fivemin.csv
Description: Cleaned five minute behavioural responses dataset (5 minutes of behaviour before and after cue exposure)
Variables
- fish_ID: fish ID
- sex: male/female
- time: before/after cue exposure
- cue: alarm cue/control cue
- tank: tank ID
- length_obs_s: length of time of observation (seconds)
- sig_s: length of time spent engaging in sigmoidal display (seconds) - males only
- sig_nb: number of sigmoidal displays - males only
- chasing_s: length of time spent chasing female (seconds) - males only
- feeding_s: length of time spent feeding (seconds)
- frozen_s: length of time spent frozen (seconds
- Q1_s: length of time in quadrant 1 (seconds)
- Q2_s: length of time in quadrant 2 (seconds)
- Q3_s: length of time in quadrant 3 (seconds)
- Q4_s: length of time in quadrant 4 (seconds)
- Q5_s: length of time in quadrant 5 (seconds)
- Q6_s: length of time in quadrant 6 (seconds)
- dash_nb: number of dashes
File: cueresponse.csv
Description: raw cue response data
Variables
- obs_id: Observation ID
- length_obs_s: length of obervation (seconds)
- sex: male/female
- sig_s: length of time spent engaging in sigmoidal displays - male only (seconds)
- sig_nb: number of sigmoidal displays - male only
- chasing_s: length of time spent chasing the female - male only (seconds)
- feeding_s: length of time spent feeding (seconds)
- frozen_s: length of time spent frozen (seconds)
- Q1_s: length of time in quadrant 1 (seconds)
- Q2_s: length of time in quadrant 2 (seconds)
- Q3_s: length of time in quadrant 3 (seconds)
- Q4_s: length of time in quadrant 4 (seconds)
- Q5_s: length of time in quadrant 5 (seconds)
- Q6_s: length of time in quadrant 6 (seconds)
- dash_nb: Number of dashes
File: cuetanks.csv
Description: raw tank and cue exposure data
Variables
- obs_id: Observation ID
- tank: tank ID
- time: before/after cue exposure
- sex: male/female
- fish_ID: fish ID
- cue: alarm cue/control
Code/software
All required packages and functions are listed at the top of each script.
Setup
1. Download project repository from Github or Dryad
2. Download data folders:
- Behavioural data: from Github or Dryad
- Sequencing data: from SRA (Short-term: SubmissionID: SUB15053745, BioProject ID: PRJNA1220975)
Code Running
All code for analysis is provided in the github repository or in scripts.zip file on Dryad. Each folder contains all scripts for a specific type of analysis (01_behavioural_analysis and 02_methylation_analysis) and the files within the folder should be run in order of numbering. Files with a and b lettering represent code paired with its slurm submission file. Note that any code with this pairing was written to be run on Compute Canada HPC system and may need to be adapted to your needs if you do not have access. All required software is commented at top of each script. Uncomment and run install lines if required. Analysis was originally run on R v4.3.2. The nf-core/methylseq pipeline v1.6.1 was used to process sequencing reads. Other details on versions of software used can be found in manuscript.
Access information
Other publicly accessible locations of the data:
- SRA: SUB15053745, BioProject ID: PRJNA1220975
We exposed guppies to alarm cue or control cue and then measured behavioural responses for 10 minutes (only 5 minutes of data used in paper) before and after cue exposure. Tanks had a back board that visually divided the tank into three equal horizontal sections so that fish position could be recorded as upper, middle, and bottom of tank. All behavioural responses to cue exposure were scored by a single observer that was blind to the treatment using BORIS v7.12.2. Behavioural videos were binned into 60 second segments for scoring. The observer recorded the time spent in each section of the tank, time spent frozen, and time spent foraging. Foraging was defined as active pecking at substrate and ended when the fish was no longer oriented towards the substrate and had not pecked for two seconds. We also recorded the time males spent pursuing females and the time males engaged in sigmoidal mating displays, a courtship display where males curve their bodies into an S-shape. After the assigned time point (0.5h, 1h, 4h, 24h, or 72h), we euthanized fish and removed brains for whole genome bisulfite sequencing.
