Evaluation of Blood–Brain Barrier Permeability and Integrity in Juvenile Rodents: Dynamic Contrast-Enhanced (DCE), Magnetic Resonance Imaging (MRI), and Evans Blue Extravasation

  • Trish Domi
  • Faraz Honarvar
  • Andrea Kassner
Part of the Neuromethods book series (NM, volume 142)


Focal ischemic stroke is the result of a blockage in an artery that leads to decreased blood flow to the neuronal cells in the brain. The middle cerebral artery (MCA) is the most common artery that is occluded in adult and pediatric stroke patients. The pathophysiology is challenging to study in either of these populations because of the highly variable clinical state in humans. Many of these variables can be eliminated when using in vivo models of stroke in rodents. Here, we describe a technique called the transient MCA occlusion (tMCAo) model in a juvenile rat model of stroke. This technique utilizes a filament that is advanced to block the origin of the MCA to induce focal ischemia. The filament is then retracted 60–90 min later allowing for secondary reperfusion. By incorporating reperfusion, this model mimics embolic strokes in humans and provides the opportunity to uncover injury associated with reflow through ischemic tissue. We are particularly interested in the reperfusion-induced injury to the blood–brain barrier (BBB) that follows after blood flow to the ischemic brain is restored.

Our goal is to provide the reader with guidelines on how to execute the tMCAo surgical procedure, with notes highlighting the advantages and limitations of the method. We also include directions on how to conduct the techniques used to evaluate the permeability of the blood–brain barrier including Evans blue extravasation, a histological procedure, and dynamic contrast-enhanced (DCE) magnetic resonance imaging (MRI), a technique used to evaluate blood–brain barrier permeability that can be applied to study stroke in a rodent model.

Key words

BBB: blood–brain barrier MCAo: middle cerebral artery occlusion tMCAo: transient middle cerebral artery occlusion stroke MRI: magnetic resonance imaging Evans Blue 



Blood–brain barrier


Common carotid artery


Dynamic contrast-enhanced imaging


Evans Blue


External carotid artery


Internal carotid artery


Middle cerebral artery


Middle cerebral artery occlusion


Revolutions per minute


Transient middle cerebral artery occlusion


Triphenyl tetrazolium chloride


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© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Division of Neurology, Department of PediatricsHospital for Sick ChildrenTorontoCanada
  2. 2.Institute of Medical Sciences, University of TorontoTorontoCanada
  3. 3.Division of Translational Medicine, Department of Medical ImagingHospital for Sick Children, University of TorontoTorontoCanada

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