Middle Cerebral Artery Occlusion
Objective: To investigate the mechanistic link between acute brain ischemia, microbiota alterations, and immune response after stroke using two distinct models of middle cerebral artery occlusion to assess lesion volume and functional outcomes
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Equipment3
Materials1
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Software1
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Protocol Steps
Middle Cerebral Artery Occlusion
Perform acute middle cerebral artery occlusion using two distinct models to induce stroke
Note: Two distinct models were used but specific details of each model are not provided in the excerpt
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”
Microbiota Analysis
Analyze gut microbiota composition using next-generation sequencing to identify dysbiosis patterns
Note: Reduced species diversity and bacterial overgrowth of bacteroidetes were identified as hallmarks of poststroke dysbiosis
View evidence from paper
“Reduced species diversity and bacterial overgrowth of bacteroidetes were identified as hallmarks of poststroke dysbiosis”
Intestinal Function Assessment
Assess intestinal barrier dysfunction and reduced intestinal motility using in vivo intestinal bolus tracking
Note: Measurements performed post-stroke
View evidence from paper
“intestinal barrier dysfunction and reduced intestinal motility as determined by in vivo intestinal bolus tracking”
Microbiota Recolonization Studies
Recolonize germ-free mice with either dysbiotic poststroke microbiota or normal control microbiota
Note: Dysbiotic microbiota recolonization exacerbates lesion volume and functional deficits compared to normal 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”
T-cell Polarization Assessment
Assess proinflammatory T-cell polarization in intestinal immune compartment and ischemic brain following 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”
Lymphocyte Migration Tracking
Track migration of intestinal lymphocytes to the ischemic brain using in vivo cell-tracking studies
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”
Fecal Microbiota Transplantation
Perform therapeutic transplantation of fecal microbiota to normalize dysbiosis and assess stroke outcome
Note: Transplantation normalizes brain lesion-induced dysbiosis and improves stroke outcome
View evidence from paper
“Therapeutic transplantation of fecal microbiota normalizes brain lesion-induced dysbiosis and improves stroke outcome”
Lesion Volume Assessment
Measure brain lesion volume as a primary outcome measure
Note: Lesion volume is affected by microbiota composition and dysbiosis status
View evidence from paper
“Recolonizing germ-free mice with dysbiotic poststroke microbiota exacerbates lesion volume and functional deficits”
Functional Outcome Assessment
Assess functional deficits and recovery following stroke and microbiota interventions
Note: Functional outcomes are improved by fecal microbiota transplantation
View evidence from paper
“Therapeutic transplantation of fecal microbiota normalizes brain lesion-induced dysbiosis and improves stroke outcome”