Distal Middle Cerebral Artery Occlusion
Objective: To investigate the effect of in vivo inhibition of miR-155 on brain recovery after experimental cerebral ischemia induced by distal middle cerebral artery occlusion (dMCAO), measuring microvasculature integrity, infarct size, and functional recovery
This is a Distal Middle Cerebral Artery Occlusion protocol using mouse as the model organism. The procedure involves 7 procedural steps, 3 equipment items, 1 materials. Extracted from a 2015 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
~3 week study window | ~48 hours hands-on
Core workflow
Distal Middle Cerebral Artery Occlusion (dMCAO) • miR-155 Inhibitor Injection • In Vivo Two-Photon Microscopy Assessment
Primary readouts
- Blood flow in peri-infarct area (two-photon microscopy)
- Microvascular integrity in peri-infarct area (two-photon microscopy)
- Capillary tight junction preservation (electron microscopy)
- Neuronal damage in peri-infarct area (electron microscopy)
Key equipment and reagents
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Protocol Steps
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Distal Middle Cerebral Artery Occlusion (dMCAO)
Surgical induction of cerebral ischemia in mice through distal middle cerebral artery occlusion
Note: This is the baseline surgical procedure to create the stroke model
View evidence from paper
“mouse distal middle cerebral artery occlusion (dMCAO)”
miR-155 Inhibitor Injection
Intravenous injection of specific miR-155 inhibitor initiated at 48 hours post-dMCAO
Note: Treatment begins in the subacute phase of stroke
View evidence from paper
“Intravenous injections of a specific miR-155 inhibitor were initiated at 48 h after mouse distal middle cerebral artery occlusion”
In Vivo Two-Photon Microscopy Assessment
Imaging of microvasculature in peri-infarct area to assess blood flow and microvascular integrity
Note: Measurements taken at multiple timepoints post-stroke
View evidence from paper
“Using in vivo two-photon microscopy, we detected improved blood flow and microvascular integrity in the peri-infarct area”
Electron Microscopy Analysis
Examination of capillary tight junctions and neuronal damage in brain tissue samples
Note: Evaluates ultrastructural preservation of tight junctions and neuronal integrity
View evidence from paper
“Electron microscopy revealed that, in contrast to the control group, these animals demonstrated well preserved capillary tight junctions”
Western Blot Analysis
Protein analysis to assess tight junction protein ZO-1 stabilization and miR-155 target protein Rheb expression
Note: Molecular analysis of mechanisms underlying improved tight junction integrity
View evidence from paper
“Western blot analysis data indicate that improved TJ integrity in the inhibitor-injected animals could be associated with stabilization of the TJ protein ZO-1”
MRI Infarct Size Measurement
Magnetic resonance imaging to quantify infarct volume and assess brain tissue preservation
Note: Primary structural outcome measure showing 34% reduction in infarct size with miR-155 inhibitor treatment
View evidence from paper
“MRI analysis showed significant (34%) reduction of infarct size in miR-155 inhibitor-injected animals at 21 d after dMCAO”
Functional Recovery Assessment
Evaluation of animal functional recovery following stroke and treatment
Note: Behavioral outcome measures reflecting preservation of brain tissue and neurological function
View evidence from paper
“Preservation of brain tissue was reflected in efficient functional recovery of inhibitor-injected animals”