Source Paper
V. Singh, S. Roth, G. Llovera, R. Sadler, D. Garzetti et al.
Journal of Neuroscience • 2016
We have identified a bidirectional communication along the brain-gut microbiota-immune axis and show that the gut microbiota is a central regulator of immune homeostasis. Acute brain lesions induced dysbiosis of the microbiome and, in turn, changes in the gut microbiota affected neuroinflammatory and functional outcome after brain injury. The microbiota impact on immunity and stroke outcome was transmissible by microbiota transplantation. Our findings support an emerging concept in which the gut microbiota is a key regulator in priming the neuroinflammatory response to brain injury. These findings highlight the key role of microbiota as a potential therapeutic target to protect brain function after injury.
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
This is a Middle Cerebral Artery Occlusion protocol using mouse as the model organism. The procedure involves 9 procedural steps, 3 equipment items, 1 materials. Extracted from a 2016 paper published in Journal of Neuroscience.
Model and subjects
mouse • Not specified in provided text • unknown • Not specified in provided text • Not specified in provided text
Study window
Estimated timing pending
Core workflow
Middle Cerebral Artery Occlusion • Microbiota Analysis • Intestinal Function Assessment
Primary readouts
Key equipment and reagents
Verified items
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Direct vendor links
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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
“Using two distinct models of acute middle cerebral artery occlusion, we show by next-generation sequencing that large stroke lesions cause gut microbiota dysbiosis”
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
“Reduced species diversity and bacterial overgrowth of bacteroidetes were identified as hallmarks of poststroke dysbiosis”
Assess intestinal barrier dysfunction and reduced intestinal motility using in vivo intestinal bolus tracking
Note: Measurements performed post-stroke
“intestinal barrier dysfunction and reduced intestinal motility as determined by in vivo intestinal bolus tracking”
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
“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 proinflammatory T-cell polarization in intestinal immune compartment and ischemic brain following microbiota recolonization
Note: Dysbiotic microbiome induces proinflammatory T-cell polarization
“recolonization of mice with a dysbiotic microbiome induces a proinflammatory T-cell polarization in the intestinal immune compartment and in the ischemic brain”
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
“Using in vivo cell-tracking studies, we demonstrate the migration of intestinal lymphocytes to the ischemic brain”
Perform therapeutic transplantation of fecal microbiota to normalize dysbiosis and assess stroke outcome
Note: Transplantation normalizes brain lesion-induced dysbiosis and improves stroke outcome
“Therapeutic transplantation of fecal microbiota normalizes brain lesion-induced dysbiosis and improves stroke outcome”
Measure brain lesion volume as a primary outcome measure
Note: Lesion volume is affected by microbiota composition and dysbiosis status
“Recolonizing germ-free mice with dysbiotic poststroke microbiota exacerbates lesion volume and functional deficits”
Assess functional deficits and recovery following stroke and microbiota interventions
Note: Functional outcomes are improved by fecal microbiota transplantation
“Therapeutic transplantation of fecal microbiota normalizes brain lesion-induced dysbiosis and improves stroke outcome”
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.
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
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
Subjects
From papermouse • Not specified in provided text • unknown • Not specified in provided text • Not specified in provided text
Cohort notes
From paperStudy included germ-free mice and mice recolonized with different microbiota
Middle Cerebral Artery Occlusion (Not specified)
Microbiota Analysis (Not specified)
Intestinal Function Assessment (Not specified)
Microbiota Recolonization Studies (Not specified)
Gut microbiota composition and diversity
From paperNext-generation sequencing used for microbiota analysis; specific statistical methods not detailed in provided excerpt
Artifact type
Endpoint measurements summarized by group or timepoint
Comparison focus
Compare endpoint magnitude between groups, timepoints, or both
Bacterial species abundance (bacteroidetes overgrowth)
From paperNext-generation sequencing used for microbiota analysis; specific statistical methods not detailed in provided excerpt
Artifact type
Endpoint measurements summarized by group or timepoint
Comparison focus
Compare endpoint magnitude between groups, timepoints, or both
Intestinal barrier function
From paperNext-generation sequencing used for microbiota analysis; specific statistical methods not detailed in provided excerpt
Artifact type
Endpoint measurements summarized by group or timepoint
Comparison focus
Compare endpoint magnitude between groups, timepoints, or both
Intestinal motility
From paperNext-generation sequencing used for microbiota analysis; specific statistical methods not detailed in provided excerpt
Artifact type
Endpoint measurements summarized by group or timepoint
Comparison focus
Compare endpoint magnitude between groups, timepoints, or both
Gut microbiota composition and diversity
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
Bacterial species abundance (bacteroidetes overgrowth)
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
Intestinal barrier function
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
Intestinal motility
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
Next-generation sequencing used for microbiota analysis; specific statistical methods not detailed in provided excerpt
Scoring or quantification
Quantify the primary readouts for this experiment: Gut microbiota composition and diversity; Bacterial species abundance (bacteroidetes overgrowth); Intestinal barrier function; Intestinal motility.
Statistical comparison
Statistical method not yet structured for this page.
Reporting output
Report representative outputs alongside summary comparisons for Gut microbiota composition and diversity, Bacterial species abundance (bacteroidetes overgrowth), Intestinal barrier function, Intestinal motility.
Source links and direct wording from the methods section for validation and deeper review.
Citation
V. Singh et al. (2016). Microbiota Dysbiosis Controls the Neuroinflammatory Response after Stroke. Journal of Neuroscience
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