The intraluminal middle cerebral artery occlusion (MCAO) model is the most frequent used model among experimental ischemic stroke models. Here we will demonstrate the entire model in detail with the guide of Laser Doppler flowmetry, and its representative results.
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Method Article
Intraluminal Middle Cerebral Artery Occlusion (MCAO) Model for Ischemic Stroke with Laser Doppler Flowmetry Guidance in Mice
* These authors contributed equally
In This Article
Summary
Abstract
Stroke is the third leading cause of death and the leading cause of disability in the world, with an estimated cost of near $70 billion in the United States in 20091,2. The intraluminal middle cerebral artery occlusion (MCAO) model was developed by Koizumi4 in 1986 to simulate this impactful human pathology in the rat. A modification of the MCAO method was later presented by Longa3. Both techniques have been widely used to identify molecular mechanisms of brain injury resulting from ischemic stroke and potential therapeutic modalities5. This relatively noninvasive method in rats has been extended to use in mice to take advantage of transgenic and knockout strains6,7. To model focal cerebral ischemia, an intraluminal suture is advanced via the internal carotid artery to occlude the base of the MCA. Retracting the suture after a specified period of time mimics spontaneous reperfusion, but the suture can also be permanently retained. This video will be demonstrating the two major approaches for performing intraluminal MCAO procedure in mice in a stepwise fashion, as well as providing insights for potential drawbacks and pitfalls. The ischemic brain tissue will subsequently be stained by 2,3,5-triphenyltetrazolium chloride (TTC) to evaluate the extent of cerebral infarction8.
Protocol
This protocol was approved by the Institutional Animal Care and Use Committee (IACUC) at the University of Florida and is in compliance with the "Principle of Laboratory Animal Care" (NIH publication No. 86-23, revised 1985).
1. Materials
- Animals: Male C57BL/6 mice (Harlan Laboratories, Indianapolis, IN, USA) weighing 20–26 grams at the time of surgery.
- Anesthesia:
- Anesthesia machine: VetEquip inhalation anesthesia system (VetEquip Inc., Pleasanton, CA, USA).
- Inhalation anesthetic: Isoflurane (Baxter Pharmaceutics, Deerfield, IL).
- Surgical tools: SuperCut Iris sc....
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Discussion
The MCAO model is an established model that replicates cerebral ischemia in murine species. This model was first used in rats and thereafter adapted to mice. Avoiding craniectomy to eliminate the influence of surgical manipulation on blood brain barrier permeability and intracranial pressure is an advantage of this technique. However, many variables, such as strain-related differences10, size of suture tip11, duration of occlusion6, body temperature12, anesthesia13
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Disclosures
Authors declare that they have no conflict of interests.
Materials
| Name | Company | Catalog Number | Comments | |
|---|---|---|---|---|
| Male C57BL/6 mice | Animal | Harlan Laboratories | 20-26 grs | |
| Dissection microscope | Microscope | Leica Microsystems | 6.3:1 zoom | |
| Fiber optic illuminator | Illuminator | TechniQuip Corp., Livermore, CA, USA | FO1–-150 | |
| Isoflurane | Anesthesia | Baxter Internationl Inc. | 1001936060 | |
| SuperCut Iris scissor, straight | Surgical tool | World Precision Instruments, Inc. | 14218 | |
| Vannas micro-scissor, straight | Surgical tool | World Precision Instruments, Inc. | 14003 | |
| Dumont fine tip forceps | Surgical tool | World Precision Instruments, Inc. | 503290 | |
| Vessel clip | Surgical tool | World Precision Instruments, Inc. | 14120 | |
| High Frequency Desiccator 900 | Surgical tool | World Precision Instruments, Inc. | 500397 | |
| 7-0 surgical Nylon monofilament suture | Suture | Ethicon Inc. | 1647G | Suture for occlusion |
| 7-0 surgical Silk suture | Suture | Ethicon Inc. | 7733G | Suture for ligation |
| Silicone RTV adhesive | Silicone rubber | World Precision Instruments, Inc. | 1571 | |
| Vetbond | Tissue adhesive | 3M | 70200746587 | |
| Puralube Vet Ointment | Eye ointment | Pharmaderm Animal Health | ||
| Buprenorphine hydrochlorode | Analgesics | Hospira Inc. | NDC 0409-2012-32 | |
| Thermal Cautery Unit | World Precision Instruments, Inc. | 501292 | ||
| PeriFlux System 5000 | TCD flowmetry | Perimed | ||
| BAT-12 | Rectal thermometer | World Precision Instruments, Inc. | BAT-12R | |
| T/PUMP, TP600 | Thermal blanket | Gaymar Industries | ||
| 2,3,5-triphenyltetrazolium chloride | Reagent | Sigma-Aldrich | T8877 | |
| Mice brain matrix slicer | Brain slicer | Zivic Instruments | BSMAS001-1 |
References
- Recommendations on stroke prevention, diagnosis, and therapy. Report of the WHO Task Force on Stroke and other Cerebrovascular Disorders. Stroke; a journal of cerebral circulation. 20, 1407-1431 (1989).
- Lloyd-Jones, D.
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