Source Paper
TGFβ signaling in the brain increases with aging and signals to astrocytes and innate immune cells in the weeks after stroke
Kristian P Doyle, Egle Cekanaviciute, Lauren E Mamer, Marion S Buckwalter
Journal of Neuroinflammation •
Source Paper
Kristian P Doyle, Egle Cekanaviciute, Lauren E Mamer, Marion S Buckwalter
Journal of Neuroinflammation •
Objective: Measure TGF-β signaling after transient focal ischemia stroke in young and old mice using real-time bioluminescence imaging over 21 days, and determine variability in bioluminescence between individual mice and effects of estrus cycle
This is a Stroke Model with Bioluminescence Monitoring protocol using mouse as the model organism. The procedure involves 8 procedural steps, 1 equipment items, 1 materials. Extracted from a 2010 paper published in Journal of Neuroinflammation.
Model and subjects
mouse • SBE-LucRT mice • unknown • 5 months old (young), 18 months old (old) • not specified • 56
Study window
~3 week study window
Core workflow
Animal cohort assignment for variability assessment • Animal cohort assignment for stroke and sham surgery groups • Transient focal ischemia stroke induction
Primary readouts
Key equipment and reagents
Verified items
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Nine females and 5 males (5 months old) were assigned to gauge variability in bioluminescence between individual mice and determine if bioluminescence is affected by the estrus cycle
Note: This cohort was used for preliminary characterization
“Nine females and 5 males (5 months old) were used to gauge variability in bioluminescence between individual mice, and to determine if bioluminescence is affected by the estrus cycle”
Eighteen young females (5 months old) and eighteen old females (18 months old) were assigned to measure TGF-β signaling after stroke. Three mice from each cohort were designated for sacrifice at days 1, 3, 7, 14 and 21 after stroke. Three mice of each age group underwent sham surgery and were imaged for 21 days prior to sacrifice
Note: Experimental paradigm was repeated in young and old male mice
“Eighteen young females (5 months old) and eighteen old females (18 months old) were used to measure TGF-β signaling after stroke in real time by bioluminescence. 3 mice from each cohort were sacrificed at days 1, 3, 7, 14 and 21 after stroke”
Stroke was induced in young and old female mice (and subsequently in male mice)
Note: Specific surgical technique not detailed in provided text
“Eighteen young females (5 months old) and eighteen old females (18 months old) were used to measure TGF-β signaling after stroke in real time by bioluminescence”
Three mice of each age group underwent sham surgery without stroke induction
Note: Sham surgery mice served as controls for uninjured brains
“Three mice of each age group also underwent sham surgery and were imaged for 21 days prior to sacrifice”
Bioluminescence imaging was performed to measure TGF-β signaling in real time over 21 days post-stroke and post-sham surgery
Note: Imaging was conducted on both stroke and sham surgery groups
“measure TGF-β signaling after stroke in real time by bioluminescence”
Three mice from each cohort were sacrificed at days 1, 3, 7, 14 and 21 after stroke for tissue analysis
Note: Tissue was collected for immunofluorescence analysis
“3 mice from each cohort were sacrificed at days 1, 3, 7, 14 and 21 after stroke”
Localization of TGF-β1 and TGF-β signaling was determined by immunofluorescence in tissue samples from stroke and sham surgery groups
Note: Performed on sacrificed tissue samples
“3 mice from each cohort were sacrificed at days 1, 3, 7, 14 and 21 after stroke for localization of TGF-β1 and TGF-β signaling by immunoflorescence”
The entire experimental paradigm (stroke induction, sham surgery, bioluminescence imaging, and tissue collection) was repeated in young and old male mice
Note: Allows comparison of sex differences in TGF-β signaling response to stroke
“This experimental paradigm was repeated in young and old male mice”
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.
Measure TGF-β signaling after transient focal ischemia stroke in young and old mice using real-time bioluminescence imaging over 21 days, and determine variability in bioluminescence between individual mice and effects of estrus cycle
Objective
Measure TGF-β signaling after transient focal ischemia stroke in young and old mice using real-time bioluminescence imaging over 21 days, and determine variability in bioluminescence between individual mice and effects of estrus cycle
Subjects
From papermouse • SBE-LucRT mice • unknown • 5 months old (young), 18 months old (old) • not specified
Sample count
From paper56
Cohort notes
From paperNine females and 5 males (5 months old) used for variability assessment; 18 young females and 18 old females for TGF-β signaling measurement; experimental paradigm repeated in young and old male mice
Animal cohort assignment for variability assessment (not specified)
Animal cohort assignment for stroke and sham surgery groups (21 days imaging period)
Transient focal ischemia stroke induction (not specified)
Sham surgery control procedure (not specified)
Real-time bioluminescence signal intensity as measure of TGF-β signaling
From papernot specified in provided text
Artifact type
Representative image panels with region or marker comparisons
Comparison focus
Compare staining intensity, structure, or cell counts across matched conditions
Variability in bioluminescence between individual mice
From papernot specified in provided text
Artifact type
Representative image panels with region or marker comparisons
Comparison focus
Compare staining intensity, structure, or cell counts across matched conditions
Effect of estrus cycle on bioluminescence
From papernot specified in provided text
Artifact type
Representative image panels with region or marker comparisons
Comparison focus
Compare staining intensity, structure, or cell counts across matched conditions
Spatial localization of TGF-β1 and TGF-β signaling by immunofluorescence
From papernot specified in provided text
Artifact type
Representative image panels with region or marker comparisons
Comparison focus
Compare staining intensity, structure, or cell counts across matched conditions
Real-time bioluminescence signal intensity as measure of TGF-β signaling
From paperRaw artifact
Field or section images captured from matched samples
Processed artifact
Selected representative panels with quantified intensity, counts, or area measurements
Final reported form
Per-group imaging summaries with representative figures and quantified endpoints
Variability in bioluminescence between individual mice
From paperRaw artifact
Field or section images captured from matched samples
Processed artifact
Selected representative panels with quantified intensity, counts, or area measurements
Final reported form
Per-group imaging summaries with representative figures and quantified endpoints
Effect of estrus cycle on bioluminescence
From paperRaw artifact
Field or section images captured from matched samples
Processed artifact
Selected representative panels with quantified intensity, counts, or area measurements
Final reported form
Per-group imaging summaries with representative figures and quantified endpoints
Spatial localization of TGF-β1 and TGF-β signaling by immunofluorescence
From paperRaw artifact
Field or section images captured from matched samples
Processed artifact
Selected representative panels with quantified intensity, counts, or area measurements
Final reported form
Per-group imaging summaries with representative figures and quantified endpoints
Acquisition
Capture matched images from the relevant tissue region using the same acquisition settings across samples.
Preprocessing / cleaning
not specified in provided text
Scoring or quantification
Quantify the primary readouts for this experiment: Real-time bioluminescence signal intensity as measure of TGF-β signaling; Variability in bioluminescence between individual mice; Effect of estrus cycle on bioluminescence; Spatial localization of TGF-β1 and TGF-β signaling by immunofluorescence.
Normalization
Normalize image-derived measurements against the matched acquisition or segmentation rules before comparing groups.
Statistical comparison
Statistical method not yet structured for this page.
Reporting output
Report representative outputs alongside summary comparisons for Real-time bioluminescence signal intensity as measure of TGF-β signaling, Variability in bioluminescence between individual mice, Effect of estrus cycle on bioluminescence, Spatial localization of TGF-β1 and TGF-β signaling by immunofluorescence.
Source links and direct wording from the methods section for validation and deeper review.
Citation
Kristian P Doyle et al. (2010). TGFβ signaling in the brain increases with aging and signals to astrocytes and innate immune cells in the weeks after stroke. Journal of Neuroinflammation
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