Animal models of ischemic stroke
Animal models of ischemic stroke are procedures inducing cerebral ischemia. The aim is the study of basic processes or potential therapeutic interventions in this disease, and the extension of the pathophysiological knowledge on and/or the improvement of medical treatment of human ischemic stroke. Ischemic stroke has a complex pathophysiology involving the interplay of many different cells and tissues such as neurons, glia, endothelium, and the immune system. These events cannot be mimicked satisfactorily in vitro yet. Thus a large portion of stroke research is conducted on animals.
- 1 Overview
- 2 Mechanisms of inducing ischemic stroke
- 3 Hypoxic Ischemia models
- 4 Focal ischemia models
- 4.1 Endothelin-1 -induced constriction of arteries and veins
- 4.2 Embolic middle cerebral artery occlusion
- 4.3 Endovascular filament middle cerebral artery occlusion
- 4.4 Permanent transcranial middle cerebral artery occlusion
- 4.5 Transient transcranial middle cerebral artery occlusion
- 4.6 Cerebrocortical photothrombosis
- 5 See also
- 6 References
- 7 References
Several models in different species are currently known to produce cerebral ischemia. Global ischemia models, both complete and incomplete, tend to be easier to perform. However, they are less immediately relevant to human stroke than the focal stroke models, because global ischemia is not a common feature of human stroke. However, in various settings global ischemia is also relevant, e.g. in global anoxic brain damage due to cardiac arrest. Different species also vary in their susceptibility to the various types of ischemic insults. An example is gerbils. They do not have a Circle of Willis and stroke can be induced by common carotid artery occlusion alone.
Mechanisms of inducing ischemic stroke
Some of the mechanisms which have been used are:
- Complete global ischemia
- Aorta/vena cava occlusion
- External neck torniquet or cuff
- Cardiac arrest
- Incomplete global ischemia
- Hemorrhage or hypotension
- Hypoxic ischemia
- Intracranial hypertension and common carotid artery occlusion
- Two-vessel occlusion and hypotension
- Four-vessel occlusion
- Unilateral common carotid artery occlusion (in some species only)
- Focal cerebral ischemia
- Endothelin-1-induced constriction of arteries and veins
- Middle cerebral artery occlusion
- Spontaneous brain infarction (in spontaneously hypertensive rats)
- Macrosphere embolization
- Multifocal cerebral ischemia
- Blood clot embolization
- Microsphere embolization
Hypoxic Ischemia models
One of the most commonly used animal models of hypoxic ischemia was originally described by Levine in 1960 and later refined by Rice et al., in 1981. This approach is useful to study hypoxic ischemia in the developing brain, since newborn rat pups are utilized in this model. Briefly, 7 day old rat pups undergo a permanent unilateral carotid artery ligation with a subsequent 3 hour exposure to a hypoxic environment (8% oxygen). This model creates a unilateral infarct in the hemisphere ipsilateral to the ligation, since the hypoxia alone is subthreshold for injury at this age. The area of injury is typically concentrated in periventricular regions of the brain, especially cortical and hippocampal areas.
Focal ischemia models
They are divided into techniques including reperfusion of the ischemic tissue (transient focal cerebral ischemia) and those without reperfusion (permanent focal cerebral ischemia). The following models are established :
- Endothelin-1 -induced constriction of arteries and veins
- Middle cerebral artery occlusion (MCAO)
- MCAO avoiding craniotomy
- Embolic middle cerebral artery occlusion
- Endovascular filament middle cerebral artery occlusion (transient or permanent)
- MCAO involving craniotomy
- Permanent transcranial middle cerebral artery occlusion
- Transient transcranial middle cerebral artery occlusion
- MCAO avoiding craniotomy
- Direct tissue damage
- Cerebrocortical photothrombosis
Endothelin-1 -induced constriction of arteries and veins
Endothelin-1 is a potent vasoconstrictor which is produced endogenously during ischemic stroke and which contributes to overall loss of cells and disability. Exogenous endothelin-1 can also be used to induce stroke and cell death after sustained vasoconstriction with reperfusion. It can be microinjected to induce focal stroke in small tissue volumes (e.g., cortical grey matter, white matter or subcortical tissue) or after injection near the Middle cerebral artery. It is often used as a model of focal stroke to evaluate candidate pro-regenerative therapies. One advantage of this model of stroke is that it causes highly reproducible infarcts. Another benefit is that it can be used in elderly rats with only very low resulting mortality.
Embolic middle cerebral artery occlusion
Middle cerebral artery (MCA) occlusion is achieved in this model by injecting particles like blood clots (thrombembolic MCAO) or artificial spheres into the carotid artery of animals as an animal model of ischemic stroke. Thrombembolic MCAO is achieved either by injecting clots that were formed in vitro or by endovascular instillation of thrombin for in situ clotting . The thrombembolic model is closest to the pathophysiology of human cardioembolic stroke. When injecting spheres into the cerebral circulation, their size determines the pattern of brain infarction: Macrospheres (300–400 µm) induce infarcts similar to those achieved by occlusion of the proximal MCA , whereas microsphere (~ 50 µm) injection results in distal, diffuse embolism . However, the quality of MCAO – and thus the volume of brain infarcts – is very variable, a fact which is further aggravated by a certain rate of spontaneous lysis of injected blood clots.
Endovascular filament middle cerebral artery occlusion
The technique of endovascular filament (intraluminal suture) MCAO as an animal model of ischemic stroke was described first by Koizumi . It is applied to rats and mice. A piece of surgical filament is introduced into the internal carotid artery and forwarded until the tip occludes the origin of the middle cerebral artery, resulting in a cessation of blood flow and subsequent brain infarction in its area of supply. If the suture is removed after a certain interval, reperfusion is achieved (transient MCAO); if the filament is left in place the procedure is suitable as model of permanent MCAO, too. The most common modification is based on Longa (1989)  who described filament introduction via the external carotid artery, allowing closure of the access point with preserved blood supply via the common and internal carotid artery to the brain after the removal of the filament. Known pitfalls of this method are insufficient occlusion, subarachnoid hemorrhage , hyperthermia , and necrosis of the ipsilateral extracranial tissue . Filament MCAO is not applicable to all rat strains .
Permanent transcranial middle cerebral artery occlusion
In this animal model of ischemic stroke the middle cerebral artery (MCA) is surgically dissected and subsequently permanently occluded, e.g. by electrocautery or ligation. Occlusion can be performed on the proximal  or distal  part of the MCA. In the latter, ischemic damage is restricted to the cerebral cortex. MCAO can be combined with temporal or permanent common carotid artery occlusion. These models require a small craniotomy.
Transient transcranial middle cerebral artery occlusion
The technique of modeling ischemic stroke by transient transcranial MCAO is similar to that of permanent transcranial MCAO, with the MCA being reperfused after a defined period of focal cerebral ischemia . Like permanent MCAO, craniotomy is required and common carotid artery (CCA) occlusion can be combined. Occluding one MCA and both CCAs is referred to as the three vessel occlusion model of focal cerebral ischemia.
Photothrombotic models of ischemic stroke use local intravascular photocoagulation of circumscribed cortical areas. After intravenous injection of photosensitive dyes like rose-bengal, the brain is irradiated through the skull via a small hole or a thinned cranial window, leading to photochemical occlusion of the irradiated vessels with secondary tissue ischemia . This approach was initially proposed by Rosenblum and El-Sabban in 1977, and improved by Watson in 1985 in the rat brain. This method has also been adapted for use in mice.
- ^ Beech, J. S., S. C. Williams, C. A. Campbell, P. M. Bath, A. A. Parsons, A. J. Hunter, D. K. Menon (2001). "Further characterisation of a thromboembolic model of stroke in the rat.". Brain Res 895 (1–2): 18–24. doi:10.1016/S0006-8993(00)03331-X. PMID 11259755.
- ^ Buchan, A. M., D. Xue, A. Slivka (February 1, 1992). "A new model of temporary focal neocortical ischemia in the rat". Stroke 23 (2): 273–9. doi:10.1161/01.str.23.2.273. PMID 1561658.
- ^ Carmichael, S. T. (2005). "Rodent models of focal stroke: size, mechanism, and purpose" ([DEAD LINK]). NeuroRx 2 (3): 396–409. doi:10.1602/neurorx.2.3.396. PMC 1144484. PMID 16389304.
- ^ Chen, S. T., C. Y. Hsu, E. L. Hogan, H. Maricq, J. D. Balentine (July 1, 1986). "A model of focal ischemic stroke in the rat: reproducible extensive cortical infarction". Stroke 17 (4): 738–43. doi:10.1161/01.str.17.4.738. PMID 2943059.
- ^ Dittmar, M., T. Spruss, G. Schuierer, M. Horn (2003). "External carotid artery territory ischemia impairs outcome in the endovascular filament model of middle cerebral artery occlusion in rats". Stroke 34 (9): 2252–7. doi:10.1161/01.STR.0000083625.54851.9A. PMID 12893948.
- ^ Dittmar, M. S., B. Vatankhah, N. P. Fehm, G. Schuierer, U. Bogdahn, M. Horn, F. Schlachetzki (2006). "Fischer-344 rats are unsuitable for the MCAO filament model due to their cerebrovascular anatomy". J Neurosci Methods 156 (1–2): 50–4. doi:10.1016/j.jneumeth.2006.02.003. PMID 16530845.
- ^ Gerriets, T., F. Li, M. D. Silva, X. Meng, M. Brevard, C. H. Sotak, M. Fisher (2003). "The macrosphere model: evaluation of a new stroke model for permanent middle cerebral artery occlusion in rats". J Neurosci Methods 122 (2): 201–11. doi:10.1016/S0165-0270(02)00322-9. PMID 12573479.
- ^ Gerriets, T., E. Stolz, M. Walberer, C. Muller, C. Rottger, A. Kluge, M. Kaps, M. Fisher, G. Bachmann (2004). "Complications and Pitfalls in Rat Stroke Models for Middle Cerebral Artery Occlusion: A Comparison Between the Suture and the Macrosphere Model Using Magnetic Resonance Angiography". Stroke 35 (10): 2372–2377. doi:10.1161/01.STR.0000142134.37512.a7. PMID 15345802.
- ^ Graham,S.M, McCullough, L.D, Murphy, S.J (2004). "Animal Models of Ischemic Stroke: Balancing Experimental Aims and Animal Care" (PDF). Comp Med 54 (5): 486–496. PMID 15575362.
- ^ Koizumi, J., Y. Yoshida, T. Nakazawa, G. Ooneda (1986). "Experimental studies of ischemic brain edema. I: a new experimental model of cerebral embolism in rats in which recirculation can be introduced in the ischemic area". Jpn J Stroke 8: 1–8.
- ^ Longa, E. Z., P. R. Weinstein, S. Carlson, R. Cummins (January 1, 1989). "Reversible middle cerebral artery occlusion without craniectomy in rats". Stroke 20 (1): 84–91. doi:10.1161/01.str.20.1.84. PMID 2643202.
- ^ Mayzel-Oreg, O., T. Omae, M. Kazemi, F. Li, M. Fisher, Y. Cohen, C. H. Sotak (2004). "Microsphere-induced embolic stroke: an MRI study". Magn Reson Med 51 (6): 1232–8. doi:10.1002/mrm.20100. PMID 15170844.
- ^ Schmid-Elsaesser, R., S. Zausinger, E. Hungerhuber, A. Baethmann, H. J. Reulen (October 1, 1989). "A critical reevaluation of the intraluminal thread model of focal cerebral ischemia: evidence of inadvertent premature reperfusion and subarachnoid hemorrhage in rats by laser-Doppler flowmetry". Stroke 29 (10): 2162–70. doi:10.1161/01.str.29.10.2162. PMID 9756599.
- ^ Tamura, A., D. I. Graham, J. McCulloch, G. M. Teasdale (1981). "Focal cerebral ischaemia in the rat: 1. Description of technique and early neuropathological consequences following middle cerebral artery occlusion". J Cereb Blood Flow Metab 1 (1): 53–60. doi:10.1038/jcbfm.1981.6. PMID 7328138.
- ^ Watson, B. D., W. D. Dietrich, R. Busto, M. S. Wachtel, M. D. Ginsberg (1985). "Induction of reproducible brain infarction by photochemically initiated thrombosis". Ann Neurol 17 (5): 497–504. doi:10.1002/ana.410170513. PMID 4004172.
- ^ Zhang, Z., R. L. Zhang, Q. Jiang, S. B. Raman, L. Cantwell, M. Chopp (1997). "A new rat model of thrombotic focal cerebral ischemia". J Cereb Blood Flow Metab 17 (2): 123–35. doi:10.1097/00004647-199702000-00001. PMID 9040491.
- Watson, Brant D.; Dietrich, W. Dalton; Busto, Raul; Wachtel, Mitchell S.; Ginsberg, Myron D. (1985). "Induction of reproducible brain infarction by photochemically initiated thrombosis". Annals of Neurology 17 (5): 497–504. doi:10.1002/ana.410170513. ISSN 0364-5134.
- Labat-gest, Vivien; Tomasi, Simone (2013). "Photothrombotic Ischemia: A Minimally Invasive and Reproducible Photochemical Cortical Lesion Model for Mouse Stroke Studies". Journal of Visualized Experiments (76). doi:10.3791/50370. ISSN 1940-087X.