Source Paper
Lauren M. Hablitz, Virginia Plá, Michael Giannetto, Hanna S. Vinitsky, Frederik Filip Stæger et al.
Nature Communications • 2020
Objective: Continuous monitoring of animal locomotor activity in 5-minute bins to analyze circadian behavioral rhythms and determine free-running periods
This is a Circadian Activity Monitoring protocol using mouse as the model organism. The procedure involves 6 procedural steps, 2 equipment items, 2 materials. Extracted from a 2020 paper published in Nature Communications.
Model and subjects
mouse • C57BL/6 and AQP4 KO • unknown • 3-5 months • 25-30g
Study window
~1.4 week study window
Core workflow
Animal acquisition and housing setup • Establish lighting conditions • Initiate continuous activity monitoring
Primary readouts
Key equipment and reagents
Verified items
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Acquire male and female C57BL/6 mice aged 3-5 months weighing 25-30g from Charles River Laboratories. Breed AQP4 KO mice in University vivarium and backcross to C57BL/6 for 20+ generations. House mice in groups with equal numbers of males and females per experimental group.
Note: Minimum of five mice per group; maintain humidity at 30-70%
“Male and female C57BL/6 mice (aged 3–5 months, weight between 25 g and 30 g) were acquired from Charles River Laboratories”
House mice either in a 12:12 light/dark cycle or under constant light conditions. For constant light experiments, house animals two per cage to reduce risk of hypothermia.
Note: Constant light housing requires two mice per cage for thermoregulation
“Mice were group-housed either in a 12:12 light/dark cycle or under constant light with ad libitum access to food and water”
Place mice in the Comprehensive Lab Animal Monitoring System (Columbus Instruments) and begin continuous monitoring of locomotor activity.
Note: Activity is recorded in 5-minute bins
“Activity was monitored continuously in 5 min bins via the Comprehensive Lab Animal Monitoring System (Columbus Instruments)”
Collect continuous activity data for at least 10 days to obtain sufficient data for circadian analysis and free-running period determination.
Note: Data collected in 5-minute bins
“Free-running period of each cage was determined using at least 10 days of activity”
Use ActogramJ software to complete circadian behavioral analysis of the collected activity data.
“Circadian behavioral analysis was completed with ActogramJ”
Apply chi-square (χ²) periodogram analysis to at least 10 days of activity data to confirm behavioral rhythmicity, estimate experimental times, and determine the free-running period of each cage in conjunction with activity onset.
Note: χ² periodogram used to confirm rhythmicity and estimate experimental timing
“Free-running period of each cage was determined using at least 10 days of activity and a χ 2 periodogram, which was used to confirm behavioral rhythmicity and estimate experimental times in conjunction with activity onset of the cage”
This section explains what the experiment is doing, which readouts matter, what the data artifacts usually look like, and how the analysis should flow from raw capture to reported result.
Continuous monitoring of animal locomotor activity in 5-minute bins to analyze circadian behavioral rhythms and determine free-running periods
Objective
Continuous monitoring of animal locomotor activity in 5-minute bins to analyze circadian behavioral rhythms and determine free-running periods
Subjects
From papermouse • C57BL/6 and AQP4 KO • unknown • 3-5 months • 25-30g
Cohort notes
From paperAQP4 KO mice backcrossed to C57BL/6 for 20+ generations; equal numbers of males and females used to control for sex differences
Animal acquisition and housing setup
Establish lighting conditions
Initiate continuous activity monitoring
Collect activity data (at least 10 days)
Locomotor activity in 5-minute bins
From paperCircadian behavioral analysis completed with ActogramJ software.
Artifact type
Longitudinal gait metrics and per-animal performance tables
Comparison focus
Compare recovery trajectory across post-injury timepoints and treatment conditions
Circadian behavioral rhythms
From paperCircadian behavioral analysis completed with ActogramJ software.
Artifact type
Endpoint measurements summarized by group or timepoint
Comparison focus
Compare endpoint magnitude between groups, timepoints, or both
Free-running period
From paperCircadian behavioral analysis completed with ActogramJ software.
Artifact type
Endpoint measurements summarized by group or timepoint
Comparison focus
Compare endpoint magnitude between groups, timepoints, or both
Behavioral rhythmicity confirmation
From paperCircadian behavioral analysis completed with ActogramJ software.
Artifact type
Endpoint measurements summarized by group or timepoint
Comparison focus
Compare endpoint magnitude between groups, timepoints, or both
Locomotor activity in 5-minute bins
From paperRaw artifact
Per-run gait capture with paw placement, timing, and stride features for each animal
Processed artifact
Cleaned gait metrics table and recovery trend summary across timepoints
Final reported form
Group comparisons of gait indices, stride metrics, or recovery curves
Circadian behavioral rhythms
From paperRaw artifact
Per-sample or per-animal endpoint measurements collected during the experiment
Processed artifact
Structured table with cleaned measurements ready for comparison
Final reported form
Summary statistics and between-group or across-timepoint comparisons
Free-running period
From paperRaw artifact
Per-sample or per-animal endpoint measurements collected during the experiment
Processed artifact
Structured table with cleaned measurements ready for comparison
Final reported form
Summary statistics and between-group or across-timepoint comparisons
Behavioral rhythmicity confirmation
From paperRaw artifact
Per-sample or per-animal endpoint measurements collected during the experiment
Processed artifact
Structured table with cleaned measurements ready for comparison
Final reported form
Summary statistics and between-group or across-timepoint comparisons
Acquisition
Collect raw experimental outputs with enough metadata to preserve sample identity, condition, and timing.
Preprocessing / cleaning
Circadian behavioral analysis completed with ActogramJ software.
Scoring or quantification
Quantify the primary readouts for this experiment: Locomotor activity in 5-minute bins; Circadian behavioral rhythms; Free-running period; Behavioral rhythmicity confirmation.
Statistical comparison
Statistical method not yet structured for this page.
Reporting output
Report representative outputs alongside summary comparisons for Locomotor activity in 5-minute bins, Circadian behavioral rhythms, Free-running period, Behavioral rhythmicity confirmation.
Source links and direct wording from the methods section for validation and deeper review.
Citation
Lauren M. Hablitz et al. (2020). Circadian control of brain glymphatic and lymphatic fluid flow. Nature Communications
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Steps
6
Evidence Quotes
10
Protocol Items
4
Linked Products
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Canonical Sync
Pending
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Computed from the current experiment record updated Feb 28, 2026.
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Steps
6
Evidence
10
Specific Products
0/0
Canonical Sync
Pending
What this score means
The verification score reflects evidence coverage, subject detail, paper provenance, step depth, and whether linked products resolve to specific item pages instead of generic searches.
Computed from the current experiment record updated Feb 28, 2026.
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