Fecal Microbiota Transplantation
Objective: Therapeutic transplantation of fecal microbiota to normalize dysbiosis and assess effects on stroke lesion volume and functional outcomes
This is a Fecal Microbiota Transplantation protocol using mouse as the model organism. The procedure involves 8 procedural steps, 3 equipment items, 1 materials. Extracted from a 2016 paper published in Journal of Neuroscience.
Model and subjects
mouse • Not explicitly stated in provided text • unknown • Not explicitly stated in provided text • Not explicitly stated in provided text
Study window
Estimated timing pending
Core workflow
Induce acute brain ischemia • Collect and analyze fecal samples • Assess intestinal barrier function and motility
Primary readouts
- Stroke lesion volume
- Functional outcomes/deficits
- Gut microbiota composition (species diversity, bacterial abundance)
- Intestinal barrier function
Key equipment and reagents
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Protocol Steps
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Induce acute brain ischemia
Perform acute middle cerebral artery occlusion using two distinct models
Note: Two distinct models of acute middle cerebral artery occlusion were used
View evidence from paper
“Using two distinct models of acute middle cerebral artery occlusion, we show by next-generation sequencing that large stroke lesions cause gut microbiota dysbiosis”
Collect and analyze fecal samples
Collect fecal microbiota from stroke-affected mice and analyze composition using next-generation sequencing
Note: Analysis identifies reduced species diversity and bacterial overgrowth of bacteroidetes as hallmarks of poststroke dysbiosis
View evidence from paper
“Reduced species diversity and bacterial overgrowth of bacteroidetes were identified as hallmarks of poststroke dysbiosis”
Assess intestinal barrier function and motility
Determine intestinal barrier dysfunction and reduced intestinal motility using in vivo intestinal bolus tracking
Note: Dysbiosis is associated with intestinal dysfunction
View evidence from paper
“intestinal barrier dysfunction and reduced intestinal motility as determined by in vivo intestinal bolus tracking”
Recolonize germ-free mice with dysbiotic microbiota
Transplant dysbiotic poststroke microbiota into germ-free mice and assess stroke outcomes
Note: Recolonization with dysbiotic microbiota exacerbates lesion volume and functional deficits compared to normal control microbiota
View evidence from paper
“Recolonizing germ-free mice with dysbiotic poststroke microbiota exacerbates lesion volume and functional deficits after experimental stroke compared with the recolonization with a normal control microbiota”
Assess immune polarization
Analyze T-cell polarization in intestinal immune compartment and ischemic brain following dysbiotic microbiota recolonization
Note: Dysbiotic microbiome induces proinflammatory T-cell polarization
View evidence from paper
“recolonization of mice with a dysbiotic microbiome induces a proinflammatory T-cell polarization in the intestinal immune compartment and in the ischemic brain”
Track intestinal lymphocyte migration
Use in vivo cell-tracking studies to demonstrate migration of intestinal lymphocytes to the ischemic brain
Note: Demonstrates bidirectional communication along brain-gut microbiota-immune axis
View evidence from paper
“Using in vivo cell-tracking studies, we demonstrate the migration of intestinal lymphocytes to the ischemic brain”
Perform therapeutic fecal microbiota transplantation
Transplant normal fecal microbiota to stroke-affected mice to normalize dysbiosis
Note: Therapeutic intervention to restore normal microbiota composition
View evidence from paper
“Therapeutic transplantation of fecal microbiota normalizes brain lesion-induced dysbiosis and improves stroke outcome”
Assess stroke outcome measures
Measure lesion volume and functional outcomes following fecal microbiota transplantation
Note: Primary outcome measures for therapeutic efficacy
View evidence from paper
“Therapeutic transplantation of fecal microbiota normalizes brain lesion-induced dysbiosis and improves stroke outcome”