An Integrated Stroke Model with a Consistent Penumbra for the Assessment of Neuroprotective Interventions

Background/Aim: A longer period of vessel occlusion reduces the coefficient of variation of the infarct lesion size [‘infarct variation coefficient' (IVC)] due to a gradual expansion of the lesion within a limited territory defined by the vascular anatomy, but it increases the mortality rate. A crucial issue in the induction of experimental focal cerebral ischemia has been to achieve a low IVC value and a low mortality rate. We attempted to improve IVC and mortality using the 3-vessel occlusion model. Methods: We introduced a new, transtemporal fascia approach to expose the zygomatic arch, in which the fascia of the temporal muscle is cut and retracted dorsally together with the facial nerve and the vein en bloc. Results/Conclusion: The approach avoided traumatic venous bleeding around the zygomatic arch. We established a bloodless model of focal ischemia that can produce a consistent degree of reduction in the regional cerebral blood flow within the ischemic penumbra. The model, characterized by a 15-min ischemia, an IVC of 15-21%, and low mortality after ischemia, is expected to produce reliable preclinical evidence in the assessment of neuroprotective interventions for ischemic stroke. The entire procedure is presented in the online supplementary video (www. karger.com/doi/10.1159/000356048).

[1]  K. Iihara,et al.  Alogliptin, a dipeptidylpeptidase-4 inhibitor, for patients with diabetes mellitus type 2, induces tolerance to focal cerebral ischemia in non-diabetic, normal mice , 2013, Brain Research.

[2]  J. Klein,et al.  Neuroprotective effects of lactate in brain ischemia: Dependence on anesthetic drugs , 2013, Neurochemistry International.

[3]  J. Klein,et al.  Blood gases and energy metabolites in mouse blood before and after cerebral ischemia: the effects of anesthetics , 2013, Experimental biology and medicine.

[4]  Andreas Meisel,et al.  Do stroke models model stroke? , 2012, Disease Models & Mechanisms.

[5]  K. Iihara,et al.  The Role of the Host Defense System in the Development of Cerebral Vasospasm: Analogies between Atherosclerosis and Subarachnoid Hemorrhage , 2012, European Neurology.

[6]  R. Morita,et al.  Peroxiredoxin family proteins are key initiators of post-ischemic inflammation in the brain , 2012, Nature Medicine.

[7]  T. Xiao,et al.  Optimizing suture middle cerebral artery occlusion model in C57BL/6 mice circumvents posterior communicating artery dysplasia. , 2012, Journal of neurotrauma.

[8]  T. Tominaga,et al.  Consistent focal cerebral ischemia without posterior cerebral artery occlusion and its real-time monitoring in an intraluminal suture model in mice. , 2012, Journal of neurosurgery.

[9]  W. Heiss The Ischemic Penumbra: Correlates in Imaging and Implications for Treatment of Ischemic Stroke , 2011, Cerebrovascular Diseases.

[10]  W. Pulsinelli,et al.  Metabolic and Perfusion Responses to Recurrent Peri-Infarct Depolarization during Focal Ischemia in the Spontaneously Hypertensive Rat: Dominant Contribution of Sporadic CBF Decrements to Infarct Expansion , 2011, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[11]  T. Miyata,et al.  NDRG4 Protein-deficient Mice Exhibit Spatial Learning Deficits and Vulnerabilities to Cerebral Ischemia* , 2011, The Journal of Biological Chemistry.

[12]  N. Hattori,et al.  Exendin-4, a glucagon-like peptide-1 receptor agonist, provides neuroprotection in mice transient focal cerebral ischemia , 2011, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[13]  J. Dreier The role of spreading depression, spreading depolarization and spreading ischemia in neurological disease , 2011, Nature Medicine.

[14]  Jing Zhang,et al.  Blood-Oxygenation-Level-Dependent-(BOLD-) Based R2′ MRI Study in Monkey Model of Reversible Middle Cerebral Artery Occlusion , 2011, Journal of biomedicine & biotechnology.

[15]  M. K. Khan,et al.  Germinated brown rice as a value added rice product: A review , 2011, Journal of food science and technology.

[16]  K. Egan,et al.  Systematic survey of the design, statistical analysis, and reporting of studies published in the 2008 volume of the Journal of Cerebral Blood Flow and Metabolism , 2010, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[17]  T. Iwaki,et al.  Pivotal role of cerebral interleukin-17–producing γδT cells in the delayed phase of ischemic brain injury , 2009, Nature Medicine.

[18]  G. A. West,et al.  A New Model of Cortical Stroke in the Rhesus Macaque , 2009, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[19]  Ulrich Dirnagl,et al.  Reprint: Good Laboratory Practice: Preventing Introduction of Bias at the Bench , 2009, Stroke.

[20]  N. Saito,et al.  Blocking pterygopalatine arterial blood flow decreases infarct volume variability in a mouse model of intraluminal suture middle cerebral artery occlusion , 2008, Journal of Neuroscience Methods.

[21]  S. Namura,et al.  Filamentous Middle Cerebral Artery Occlusion Causes Ischemic Damage to the Retina in Mice , 2008, Stroke.

[22]  O. Sakowitz,et al.  Spreading depolarizations occur in human ischemic stroke with high incidence , 2008, Annals of neurology.

[23]  T. Krings,et al.  Cerebral Proliferative Angiopathy: Clinical and Angiographic Description of an Entity Different From Cerebral AVMs , 2008, Stroke.

[24]  S. Savitz A critical appraisal of the NXY-059 neuroprotection studies for acute stroke: A need for more rigorous testing of neuroprotective agents in animal models of stroke , 2007, Experimental Neurology.

[25]  S. Warach,et al.  Recommendations from the STAIR V meeting on acute stroke trials, technology and outcomes. , 2007, Stroke.

[26]  A. Dunn,et al.  Peri-infarct depolarizations lead to loss of perfusion in ischaemic gyrencephalic cerebral cortex. , 2006, Brain : a journal of neurology.

[27]  G. Donnan,et al.  1,026 Experimental treatments in acute stroke , 2006, Annals of neurology.

[28]  MarcFisher Enhancing the Development and Approval of Acute Stroke Therapies , 2005 .

[29]  T. Lüscher,et al.  Aggravation of Focal Cerebral Ischemia by Tissue Plasminogen Activator Is Reversed by 3-Hydroxy-3-Methylglutaryl Coenzyme A Reductase Inhibitor but Does Not Depend on Endothelial NO Synthase , 2005, Stroke.

[30]  Kenichi Murao,et al.  Spreading depression induces long-lasting brain protection against infarcted lesion development via BDNF gene-dependent mechanism , 2004, Brain Research.

[31]  P. Barber,et al.  Temperature-Regulated Model of Focal Ischemia in the Mouse: A Study With Histopathological and Behavioral Outcomes , 2004, Stroke.

[32]  Ulrich Dirnagl,et al.  Selective Neuronal Vulnerability Following Mild Focal Brain Ischemia in the Mouse , 2003, Brain pathology.

[33]  I. Nagata,et al.  Evaluation of MCAO stroke models in normotensive rats: standardized neocortical infarction by the 3VO technique , 2003, Experimental Neurology.

[34]  M. Ginsberg,et al.  Adventures in the Pathophysiology of Brain Ischemia: Penumbra, Gene Expression, Neuroprotection: The 2002 Thomas Willis Lecture , 2003, Stroke.

[35]  A. Tamura,et al.  Differences in the Extent of Primary Ischemic Damage between Middle Cerebral Artery Coagulation and Intraluminal Occlusion Models , 2002, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[36]  M. Ninomiya,et al.  Protective Effect of Endothelin Type A Receptor Antagonist on Brain Edema and Injury After Transient Middle Cerebral Artery Occlusion in Rats , 2001, Stroke.

[37]  I. Nakahara,et al.  Combination of intraischemic and postischemic hypothermia provides potent and persistent neuroprotection against temporary focal ischemia in rats. , 1999, Stroke.

[38]  T. Back Pathophysiology of the Ischemic Penumbra—Revision of a Concept , 1998, Cellular and Molecular Neurobiology.

[39]  H. Reulen,et al.  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. , 1998, Stroke.

[40]  W J Powers,et al.  Hemodynamic Effects of Middle Cerebral Artery Stenosis and Occlusion , 1998, AJNR. American journal of neuroradiology.

[41]  M. Hori,et al.  Cerebral Ischemia after Bilateral Carotid Artery Occlusion and Intraluminal Suture Occlusion in Mice: Evaluation of the Patency of the Posterior Communicating Artery , 1998, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[42]  M. Moskowitz,et al.  Reduced fertility and postischaemic brain injury in mice deficient in cytosolic phospholipase A2 , 1997, Nature.

[43]  R. Busto,et al.  Transient Middle Cerebral Artery Occlusion by Intraluminal Suture: I. Three-Dimensional Autoradiographic Image-Analysis of Local Cerebral Glucose Metabolism—Blood Flow Interrelationships during Ischemia and Early Recirculation , 1997, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[44]  J. Baron,et al.  Early reperfusion in the anesthetized baboon reduces brain damage following middle cerebral artery occlusion: a quantitative analysis of infarction volume. , 1997, Stroke.

[45]  KEVIN S. Lee,et al.  Mild postischemic hypothermia limits cerebral injury following transient focal ischemia in rat neocortex , 1996, Brain Research.

[46]  K. Sakatani,et al.  Isotonic mannitol and the prevention of local heat generation and tissue adherence to bipolar diathermy forceps tips during electrical coagulation. Technical note. , 1995, Journal of neurosurgery.

[47]  K. Hossmann Viability thresholds and the penumbra of focal ischemia , 1994, Annals of neurology.

[48]  G. Unsgård,et al.  Characterization of the microcirculation during ischemia and reperfusion in the penumbra of a rat model of temporary middle cerebral artery occlusion: a laser Doppler flowmetry study. , 1994, International journal of microcirculation, clinical and experimental.

[49]  Jun Chen,et al.  Human Copper‐Zinc Superoxide Dismutase Transgenic Mice Are Highly Resistant to Reperfusion Injury After Focal Cerebral Ischemia , 1994, Stroke.

[50]  K. Hossmann,et al.  Cortical Negative DC Deflections following Middle Cerebral Artery Occlusion and KCl-Induced Spreading Depression: Effect on Blood Flow, Tissue Oxygenation, and Electroencephalogram , 1994, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[51]  Y. Ushio,et al.  Changes of immunoreactivity for synaptophysin (‘protein p38’) following a transient cerebral ischemia in the rat striatum , 1993, Brain Research.

[52]  W. Pulsinelli,et al.  The CBF Threshold and Dynamics for Focal Cerebral Infarction in Spontaneously Hypertensive Rats , 1992, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[53]  W. Kofke,et al.  Comparison of the effects of volatile anesthetics in varying concentrations on brain energy metabolism with brain ischemia in rats , 1992, Neurochemical Research.

[54]  W. Pulsinelli,et al.  Focal Brain Ischemia in the Rat: Methods for Reproducible Neocortical Infarction Using Tandem Occlusion of the Distal Middle Cerebral and Ipsilateral Common Carotid Arteries , 1988, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[55]  L. Pitts,et al.  Evaluation of 2,3,5-triphenyltetrazolium chloride as a stain for detection and quantification of experimental cerebral infarction in rats. , 1986, Stroke.

[56]  E. Hogan,et al.  A model of focal ischemic stroke in the rat: reproducible extensive cortical infarction. , 1986, Stroke.

[57]  L. Pitts,et al.  Rat middle cerebral artery occlusion: evaluation of the model and development of a neurologic examination. , 1986, Stroke.

[58]  M. Raichle,et al.  Cerebral Blood Flow and Cerebral Metabolic Rate of Oxygen Requirements for Cerebral Function and Viability in Humans , 1985, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[59]  W. Calder,et al.  Body size, mortality, and longevity. , 1983, Journal of theoretical biology.

[60]  B. Siesjö,et al.  Thresholds in cerebral ischemia - the ischemic penumbra. , 1981, Stroke.

[61]  D. Graham,et al.  Focal Cerebral Ischaemia in the Rat: 1. Description of Technique and Early Neuropathological Consequences following Middle Cerebral Artery Occlusion , 1981, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[62]  N M Branston,et al.  Cortical Evoked Potential and Extracellular K+ and H+ at Critical Levels of Brain Ischemia , 1977, Stroke.

[63]  H. Winn,et al.  RODENT STROKE MODEL GUIDELINES FOR PRECLINICAL STROKE TRIALS (1ST EDITION). , 2009, Journal of experimental stroke & translational medicine.

[64]  T. Kirino,et al.  Proximal occlusion of the middle cerebral artery in C57Black6 mice: relationship of patency of the posterior communicating artery, infarct evolution, and animal survival. , 2004, Journal of neurosurgery.

[65]  B. Siesjö,et al.  Ischemic penumbra in a model of reversible middle cerebral artery occlusion in the rat , 2004, Experimental Brain Research.

[66]  Z. Zhang,et al.  Ischemia Prolonged Mild Hypothermia Therapy Protects the Brain Against Permanent Focal , 2000 .

[67]  C H Rabb,et al.  Nylon monofilament for intraluminal middle cerebral artery occlusion in rats. , 1996, Stroke.

[68]  N. Lassen,et al.  Ischemic Penumbra and Neuronal Death: Comments on the Therapeutic Window in Acute Stroke with Particular Reference to Thrombolytic Therapy , 1991 .

[69]  S. Finklestein,et al.  Increased levels of basic fibroblast growth factor (bFGF) following focal brain injury. , 1990, Restorative neurology and neuroscience.

[70]  P. Weinstein,et al.  Reversible middle cerebral artery occlusion without craniectomy in rats. , 1989, Stroke.

[71]  Yoji Yoshida,et al.  Experimental studies of ischemic brain edema , 1986 .