Controlled Cortical Impact
Objective: To assess the contribution of MMP-9 to traumatic brain injury pathophysiology by comparing morphological and motor outcomes between MMP-9 knock-out mice and wild-type littermates following controlled cortical impact
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Protocol Steps
Controlled Cortical Impact Injury
Perform controlled cortical impact procedure on mice to induce traumatic brain injury
Note: Procedure performed on both MMP-9 knock-out and wild-type littermate mice
View evidence from paper
“After controlled cortical impact in mice, MMP-9 was increased in traumatized brain”
Motor Assessment at 1 Day Post-Injury
Measure motor outcomes using rotarod device at 1 day after traumatic brain injury
Note: First timepoint for motor assessment
View evidence from paper
“Motor outcomes were measured at 1, 2, and 7 d after traumatic brain injury by the use of a rotarod device”
Motor Assessment at 2 Days Post-Injury
Measure motor outcomes using rotarod device at 2 days after traumatic brain injury
Note: Second timepoint for motor assessment
View evidence from paper
“Motor outcomes were measured at 1, 2, and 7 d after traumatic brain injury by the use of a rotarod device”
Motor Assessment at 7 Days Post-Injury
Measure motor outcomes using rotarod device at 7 days after traumatic brain injury
Note: Final timepoint for motor assessment; lesion volume analysis also performed at this timepoint
View evidence from paper
“Motor outcomes were measured at 1, 2, and 7 d after traumatic brain injury by the use of a rotarod device”
Brain Tissue Collection and Histological Processing
Collect brain tissue and prepare Nissl-stained histological sections for lesion volume measurement
Note: Performed at 7 day timepoint
View evidence from paper
“At 7 d, traumatic brain lesion volumes on Nissl-stained histological sections were significantly smaller in MMP-9 knock-out mice”
MMP-9 Level Assessment at 3 Hours Post-Injury
Measure MMP-9 levels using zymography at 3 hours after traumatic brain injury
Note: Earliest timepoint showing MMP-9 elevation
View evidence from paper
“Zymograms showed that MMP-9 was elevated as early as 3 hr after traumatic brain injury”
MMP-9 Level Assessment at 24 Hours Post-Injury
Measure total MMP-9 levels using substrate cleavage assay and zymography at 24 hours after traumatic brain injury
Note: Peak MMP-9 levels observed at this timepoint
View evidence from paper
“Total MMP-9 levels at 24 hr were significantly increased as measured by a substrate cleavage assay. Zymograms showed that MMP-9 was elevated as early as 3 hr after traumatic brain injury, reaching a maximum at ≈24 hr”
MMP-9 Level Assessment up to 1 Week Post-Injury
Monitor MMP-9 levels through 1 week post-injury using Western blot analysis and zymography
Note: Elevated MMP-9 levels persist throughout this period
View evidence from paper
“Increased MMP-9 levels persisted for up to 1 week. Western blot analysis indicated increased profiles of MMP-9 expression that corresponded with the zymographic data”