Source Paper
Fecal microbiota transplantation protects rotenone-induced Parkinson’s disease mice via suppressing inflammation mediated by the lipopolysaccharide-TLR4 signaling pathway through the microbiota-gut-brain axis
Zhe Zhao, Jingwen Ning, Xiu-qi Bao, Meiyu Shang, Jingwei Ma et al.
Microbiome • 2021
Intestinal Transit Test
Objective: Measurement of intestinal transit distance to assess gastrointestinal motility in mice
Protocol Steps
Animal acclimation
Mice were acclimated to standard laboratory conditions with 12-hour light/dark cycle, temperature 22±2°C, humidity 50-60%, with ad libitum access to food and water
Note: Humidity maintained at 50-60%. All procedures followed guidelines from Beijing Municipal Ethics Committee
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“The mice were then acclimatized (12-h light/dark cycle) under standard conditions (temperature 22 ± 2 °C, humidity 50–60%) with ad libitum access to food and water for 7 days”
Baseline group assignment
45 mice randomly assigned into two groups: control group (n=15) and model group (n=30)
Note: Random assignment performed at beginning of study
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“A total of 45 mice were randomly assigned into two groups: the control group (n = 15) and the model group (n = 30)”
Rotenone treatment phase
Model group received daily oral administration of rotenone. Control group received vehicle administration daily
Note: This phase establishes the Parkinson's disease-like model
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“In the beginning 4 weeks, the model group received the oral administration of rotenone every day. Meanwhile, the control group mice were administrated with vehicle”
Secondary group assignment
After 4 weeks, model group mice were randomly divided into two groups: Rotenone group (n=15) and FMT group (n=15)
Note: Control group remained unchanged
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“After 4 weeks, we randomly divided the model group mice into two groups: Rotenone group (n = 15) and FMT group (n = 15)”
FMT treatment phase
FMT group received fecal microbiota transplant once per day. Control group and Rotenone group received vehicle administration once per day
Note: Treatment administered during weeks 5 and 6 of the study
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“During week 5 and 6, the mice in the FMT group were treated with FMT once per day. In the meantime, the control group and the rotenone group mice received vehicle administration once a day”
Body weight monitoring
All mice were weighed daily throughout the study period
Note: Weight measurements recorded from week 1 through week 6
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“These mice were daily weighed for 6 weeks”
Intestinal transit test
Measurement of intestinal transit distance to assess gastrointestinal motility. Test performed at week 6
Note: n=5 replicates in each group for this measurement
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“GI function tests and behavioral tests were performed at week 6”
Additional behavioral and GI function tests
Rota-Rod test, Adhesive removal test, Grip strength test, Pole test, Colon length measurement, Fecal water percentage analysis, and Fecal output measurement performed at week 6
Note: n=15 for behavioral tests and fecal analysis; n=5 replicates for intestinal transit and colon length
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“GI function tests and behavioral tests were performed at week 6”
Animal sacrifice and tissue collection
All mice were sacrificed at week 6 for further analysis
Note: Tissues collected for downstream analysis
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“All the mice were sacrificed at week 6 for further analysis”