Source Paper
Zhigang Liu, Xiaoshuang Dai, Hongbo Zhang, Renjie Shi, Yan Hui et al.
Nature Communications • 2020
Abstract Cognitive decline is one of the complications of type 2 diabetes (T2D). Intermittent fasting (IF) is a promising dietary intervention for alleviating T2D symptoms, but its protective effect on diabetes-driven cognitive dysfunction remains elusive. Here, we find that a 28-day IF regimen for diabetic mice improves behavioral impairment via a microbiota-metabolites-brain axis: IF enhances mitochondrial biogenesis and energy metabolism gene expression in hippocampus, re-structures the gut microbiota, and improves microbial metabolites that are related to cognitive function. Moreover, strong connections are observed between IF affected genes, microbiota and metabolites, as assessed by integrative modelling. Removing gut microbiota with antibiotics partly abolishes the neuroprotective effects of IF. Administration of 3-indolepropionic acid, serotonin, short chain fatty acids or tauroursodeoxycholic acid shows a similar effect to IF in terms of improving cognitive function. Together, our study purports the microbiota-metabolites-brain axis as a mechanism that can enable therapeutic strategies against metabolism-implicated cognitive pathophysiologies.
Objective: Cognitive behavioral assessment to evaluate spatial learning and memory in mice using water maze test
This is a Water Maze Test protocol using mouse as the model organism. The procedure involves 9 procedural steps, 1 equipment items, 9 materials. Extracted from a 2020 paper published in Nature Communications.
Model and subjects
mouse • BKS.Cg-Dock7m+/+Leprdb/J and heterozygous controls • male • 4 months at start of intervention
Study window
~4 week study window | ~24 hours hands-on
Core workflow
Animal housing and acclimation • Baseline feeding and monitoring • Intermittent fasting regimen initiation
Primary readouts
Key equipment and reagents
Verified items
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Male BKS.Cg-Dock7m+/+Leprdb/J mice obtained from The Jackson Laboratory and housed in animal facility under standard conditions
Note: Strict 12h:12h light:dark cycle maintained; humidity at 50 ± 15%
“All mice were originally obtained from The Jackson Laboratory (Bar Harbor, ME) and housed in the Northwest A&F University animal facility under standard conditions with a strict 12 h:12 h light:dark cycle, humidity at 50 ± 15%, temperature 22 ± 2°C”
Mice fed ad libitum with regular chow (AIN-93M) and pure water before intervention at 4 months of age
Note: Bodyweight, food intake, and water consumption recorded on fasting day
“The animals were fed ad libitum before the IF was initiated at 4 months of age. Mice were fed a regular chow (AIN-93M, purchased from TROPHIC Animal Feed High-tech Co. Ltd, Nantong, China) and pure water. Bodyweight, food intake, and water consumption were recorded on fasting day”
Mice divided into groups and subjected to intermittent fasting protocol: food deprivation for 24 hours every other day with ad libitum feeding on intervening days
Note: Applied to db/m and db/db mice in multiple cohorts
“the IF mice were deprived of food for 24 h, every other day and were fed ad libitum on the intervening day for 28 days”
Antibiotics cocktail administered in drinking water to deplete gut microbiota in designated groups
Note: Different concentrations used for db/db mice (full strength) and db/m mice (half concentration)
“The antibiotics cocktail...were given in the drinking water starting 14 days before the IF regimen and throughout the experiment”
For Set 4 animals, IPA, 5-HT, and TUDCA administered via intraperitoneal injection; SCFAs dissolved in drinking water
Note: Control groups received saline injections of same volume
“For db/db-IPA, db/db-5-HT, db/db-TUDCA groups, mice were intraperitoneally injected with IPA (10 mg kg−1 d−1), 5-HT (1 mg kg−1 d−1), and TUDCA (250 mg kg−1 d−1) dissolved in saline, respectively. The mice in other groups were injected with same volume saline”
16S rRNA copies in feces detected via qPCR to confirm antibiotics effectiveness
“The 16S rRNA copies in the feces of the animals after antibiotics treatment were detected with qPCR”
Cognitive behavioral assessment performed to evaluate spatial learning and memory
Note: All behavioral studies conducted on fasting day of IF regimen. One mouse in db/db group drowned during test and data excluded
“After IF regimen, the antibiotics treatment, and metabolites treatment, the cognitive behavioral assessment was determined with a water-maze test. All of the behavioral studies and biochemical samples were collected/detected on the fasting day of IF regimen”
After behavioral testing, mice euthanized and serum and tissue samples collected
Note: Samples either snap-frozen in liquid nitrogen and stored at -80°C or stored in 4% paraformaldehyde for histological analysis
“After behavioral tests, mice were killed, then the serum and tissues samples were collected by either snap-frozen by liquid nitrogen and store at −80°C or directly stored in 4% paraformaldehyde for histological analysis”
Fecal, serum, and hippocampus samples analyzed for gut microbiome, metabolome, and RNA sequencing
“Fecal, serum, and hippocampus samples were collected to analyze gut microbiome, metabolome, and RNA sequencing, respectively”
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.
Cognitive behavioral assessment to evaluate spatial learning and memory in mice using water maze test
Objective
Cognitive behavioral assessment to evaluate spatial learning and memory in mice using water maze test
Subjects
From papermouse • BKS.Cg-Dock7m+/+Leprdb/J and heterozygous controls • male • 4 months at start of intervention
Cohort notes
From paperHomozygous Leprdb/db mice were diabetic; heterozygous Leprdb/m mice were used as controls.
Animal housing and acclimation
Baseline feeding and monitoring
Intermittent fasting regimen initiation (28 days)
Antibiotics treatment initiation (Starting 14 days before IF regimen and throughout experiment)
Spatial learning and memory performance in water maze test
From paper16S rRNA copies detected via qPCR; plasma metabolomics analyzed with two samples excluded due to poor measurement quality
Artifact type
Endpoint measurements summarized by group or timepoint
Comparison focus
Compare endpoint magnitude between groups, timepoints, or both
Bodyweight changes
From paper16S rRNA copies detected via qPCR; plasma metabolomics analyzed with two samples excluded due to poor measurement quality
Artifact type
Endpoint measurements summarized by group or timepoint
Comparison focus
Compare endpoint magnitude between groups, timepoints, or both
Food intake
From paper16S rRNA copies detected via qPCR; plasma metabolomics analyzed with two samples excluded due to poor measurement quality
Artifact type
Endpoint measurements summarized by group or timepoint
Comparison focus
Compare endpoint magnitude between groups, timepoints, or both
Water consumption
From paper16S rRNA copies detected via qPCR; plasma metabolomics analyzed with two samples excluded due to poor measurement quality
Artifact type
Endpoint measurements summarized by group or timepoint
Comparison focus
Compare endpoint magnitude between groups, timepoints, or both
Spatial learning and memory performance in water maze test
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
Bodyweight changes
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
Food intake
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
Water consumption
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
16S rRNA copies detected via qPCR; plasma metabolomics analyzed with two samples excluded due to poor measurement quality
Scoring or quantification
Quantify the primary readouts for this experiment: Spatial learning and memory performance in water maze test; Bodyweight changes; Food intake; Water consumption.
Statistical comparison
Statistical method not yet structured for this page.
Reporting output
Report representative outputs alongside summary comparisons for Spatial learning and memory performance in water maze test, Bodyweight changes, Food intake, Water consumption.
Source links and direct wording from the methods section for validation and deeper review.
Citation
Zhigang Liu et al. (2020). Gut microbiota mediates intermittent-fasting alleviation of diabetes-induced cognitive impairment. Nature Communications
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TROPHIC Animal Feed High-tech Co. Ltd • AIN-93M
1 item with ReplicateScience direct pages
Estimated: $690.00
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