Abstract
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.
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Abbreviations
- BBB:
-
Blood–brain barrier
- CCA:
-
Common carotid artery
- DCE:
-
Dynamic contrast-enhanced imaging
- EB:
-
Evans Blue
- ECA:
-
External carotid artery
- ICA:
-
Internal carotid artery
- MCA:
-
Middle cerebral artery
- MCAo:
-
Middle cerebral artery occlusion
- RPM:
-
Revolutions per minute
- tMCAo:
-
Transient middle cerebral artery occlusion
- TTC:
-
Triphenyl tetrazolium chloride
References
Engel O, Kolodziej S, Dirnagl U, Prinz V (2011) Modeling stroke in mice - middle cerebral artery occlusion with the filament model. J Vis Exp 47:e2423. https://doi.org/10.3791/2423
Tsze DS, Valente JH (2011) Pediatric stroke: a review. Emerg Med Int 2011:1–10. https://doi.org/10.1155/2011/734506
Uluç K, Miranpuri A, Kujoth GC, Aktüre E, Başkaya MK (2011) Focal cerebral ischemia model by endovascular suture occlusion of the middle cerebral artery in the rat. J Vis Exp (48). doi:https://doi.org/10.3791/1978
Kawamura S, Shirasawa M, Fukasawa H, Yasui N (1991) Attenuated neuropathology by nilvadipine after middle cerebral artery occlusion in rats. Stroke 22(1):51–55. Retrieved from http://www.ncbi.nlm.nih.gov/pubmed/1987673
Takano K, Tatlisumak T, Bergmann AG, Gibson DG, Fisher M (1997) Reproducibility and reliability of middle cerebral artery occlusion using a silicone-coated suture (Koizumi) in rats. J Neurol Sci 153(1):8–11. Retrieved from http://www.ncbi.nlm.nih.gov/pubmed/9455971
Carmichael ST (2005) Rodent models of focal stroke: size, mechanism, and purpose. NeuroRx 2(3):396–409. https://doi.org/10.1602/neurorx.2.3.396
Langheinrich AC, Yeniguen M, Ostendorf A, Marhoffer S, Kampschulte M, Bachmann G et al (2010) Evaluation of the middle cerebral artery occlusion techniques in the rat by in-vitro 3-dimensional micro- and nano computed tomography. BMC Neurol 10:36. https://doi.org/10.1186/1471-2377-10-36
Longa EZ, Weinstein PR, Carlson S, Cummins R (1989) Reversible middle cerebral artery occlusion without craniectomy in rats. Stroke 20(1):84–91 Retrieved from http://www.ncbi.nlm.nih.gov/pubmed/2643202
Garcia JH, Wagner S, Liu KF, Hu XJ (1995) Neurological deficit and extent of neuronal necrosis attributable to middle cerebral artery occlusion in rats. Statistical validation. Stroke 26(4):627–634; discussion 635. Retrieved from http://www.ncbi.nlm.nih.gov/pubmed/7709410
Merali Z, Huang K, Mikulis D, Silver F, Kassner A (2017) Evolution of blood-brain-barrier permeability after acute ischemic stroke. PLoS One 12(2):e0171558. https://doi.org/10.1371/journal.pone.0171558
Patlak CS, Blasberg RG (1985) Graphical evaluation of blood-to-brain transfer constants from multiple-time uptake data. Generalizations. J Cereb Blood Flow Metab 5(4):584–590. https://doi.org/10.1038/jcbfm.1985.87
Ewing JR, Knight RA, Nagaraja TN, Yee JS, Nagesh V, Whitton PA et al (2003) Patlak plots of Gd-DTPA MRI data yield blood-brain transfer constants concordant with those of 14C-sucrose in areas of blood-brain opening. Magn Reson Med 50(2):283–292. https://doi.org/10.1002/mrm.10524
Merali Z, Leung J, Mikulis D, Silver F, Kassner A (2015) Longitudinal assessment of imatinib’s effect on the blood-brain barrier after ischemia/reperfusion injury with permeability MRI. Transl Stroke Res 6(1):39–49. https://doi.org/10.1007/s12975-014-0358-6
Canazza A, Minati L, Boffano C, Parati E, Binks S (2014) Experimental models of brain ischemia: a review of techniques, magnetic resonance imaging, and investigational cell-based therapies. Front Neurol 5:1–15. https://doi.org/10.3389/fneur.2014.00019
Mhairi Macrae I (1992) New models of focal cerebral ischaemia. Br J Clin Pharmacol 34(4):302–308. Retrieved from http://www.ncbi.nlm.nih.gov/pubmed/1457262
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Domi, T., Honarvar, F., Kassner, A. (2019). Evaluation of Blood–Brain Barrier Permeability and Integrity in Juvenile Rodents: Dynamic Contrast-Enhanced (DCE), Magnetic Resonance Imaging (MRI), and Evans Blue Extravasation. In: Barichello, T. (eds) Blood-Brain Barrier. Neuromethods, vol 142. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8946-1_17
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DOI: https://doi.org/10.1007/978-1-4939-8946-1_17
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