腔内中东大脑中动脉闭塞（MCAO）缺血性中风模型在小鼠体内的激光多普勒血流指导
Summary
腔内大脑中动脉闭塞（MCAO）模型是最常见的缺血性中风的实验模型中使用的模型。在这里，我们将展示在整个模型用激光多普勒血流计指南的细节，其代表性的成果。
Abstract
中风是第三大死因，在世界上的残疾的首要原因，美国在2009 年 1,2的近70亿美元，估计所需费用。腔内大脑中动脉闭塞（MCAO）模型，于1986年，以模拟在大鼠有影响力的人类病理学由小泉纯一郎 4 。缺血法的修改后由龙格 3 。这两种技术已被广泛用来识别脑损伤的分子机制，导致缺血性中风和潜在的治疗方式5。一直延续这相对无创的方法，在大鼠，小鼠的使用，利用转基因和基因敲除株6,7优势。局灶性脑缺血模型，线栓是先进的经颈内动脉闭塞MCA的基地。回缩后，在指定的时间内缝合模仿自发再灌注损伤，但缝合，也可永久保留。该视频将展示在小鼠进行腔内缺血过程在逐步时尚的两个主要方法，以及为潜在的弊端和缺陷的见解。缺血性脑组织也将随之由2,3,5 -三苯基氯化四氮唑（TTC）染色，评估脑梗塞 8的程度。
Protocol
该协议被批准的机构动物护理和使用委员会（IACUC）在佛罗里达大学，并遵守的“实验动物保健的原则”（美国国立卫生研究院出版号86-23，1985年修订）。 1。材料动物：雄性C57BL / 6小鼠（哈伦实验室，印第安纳波利斯，在美国）在手术时重达20-26克。 麻醉： 麻醉机：VetEquip吸入麻醉系统（VetEquip公司，Pleasanton的，CA，美国）。 吸入麻醉药异氟醚（?…
Discussion
MCAO模型是一个既定的模式，复制脑缺血小鼠物种。第一次使用这个模型是在大鼠和以后适应小鼠。避免去骨瓣，以消除对血脑屏障通透性和颅内压手术操作的影响，是该技术的优点。然而，许多变量，如应变相关的差异 10，大小缝合技巧 11，持续时间 6闭塞， 体温 12， 麻醉 13，以及其他因素，可能导致梗塞体积在这个模型不一致。
值得一提的有具?…
Divulgazioni
The authors have nothing to disclose.
Materials
| Name of the reagent | Tipo | Company | Catalogue number | Comments (optional) |
|---|---|---|---|---|
| Male C57BL/6 mice | Animal | Harlan Laboratories, Indianapolis, IN, USA | 20-26 grs | |
| Dissection microscope | Microscope | Leica stereo Microscope MZ6 | 6.3:1 zoom | |
| Fiber optic illuminator | Illuminator | TechniQuip Corp., Livermore, CA, USA | FO1–150 | |
| Isoflurane | Anesthesia | Baxter Pharmaceutics, Deerfield, IL | 1001936060 | |
| SuperCut Iris scissor, straight | Surgical tool | World Precision Instruments, Inc., Sarasota, FL, USA | 14218 | |
| Vannas micro-scissor, straight | Surgical tool | World Precision Instruments, Inc., Sarasota, FL, USA | 14003 | |
| Dumont fine tip forceps | Surgical tool | World Precision Instruments, Inc., Sarasota, FL, USA | 503290 | |
| Vessel clip | Surgical tool | World Precision Instruments, Inc., Sarasota, FL, USA | 14120 | |
| High Frequency Desiccator 900 | Surgical tool | World Precision Instruments, Inc., Sarasota, FL, USA | 500397 | |
| 7-0 surgical Nylon monofilament suture | Suture | Ethicon, Inc., Somerville, NJ, USA | 1647G | Suture for occlusion |
| 7-0 surgical Silk suture | Suture | Ethicon, Inc., Somerville, NJ, USA | 7733G | Suture for ligation |
| Silicone RTV adhesive | Silicone rubber | World Precision Instruments, Inc., Sarasota, FL, USA | 1571 | |
| Vetbond | Tissue adhesive | 3M Company, St Paul, MN, USA | 70200746587 | |
| Puralube Vet Ointment | Eye ointment | Pharmaderm, Melville, NY, USA | ||
| Buprenorphine hydrochlorode | Analgesics | Hospira Inc., Lake Forest, IL, USA | NDC 0409-2012-32 | |
| Thermal Cautery Unit | World Precision Instruments, Inc., Sarasota, FL, USA | 501292 | ||
| PeriFlux System 5000 | TCD flowmetry | Perimed, Stockholm, Sweeden | ||
| BAT-12 | Rectal thermometer | World Precision Instruments, Inc., Sarasota, FL, USA | BAT-12R | |
| T/PUMP, TP600 | Thermal blanket | Gaymar, NY, USA | ||
| 2,3,5-triphenyltetrazolium chloride | Reagent | Sigma-Aldrich Co., St Louis, MO, USA | T8877 | |
| Mice brain matrix slicer | Brain slicer | Zivic-Miller Lab., Inc., Allison Park, PA, USA | BSMAS001-1 |
Riferimenti
- . 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. Heart disease and stroke statistics–2009 update: a report from the American Heart Association Statistics Committee and Stroke Statistics Subcommittee. Circulation. 119, e21-e181 (2009).
- Longa, E. Z., Weinstein, P. R., Carlson, S., Cummins, R. Reversible middle cerebral artery occlusion without craniectomy in rats. Stroke; a journal of cerebral circulation. 20, 84-91 (1989).
- Koizumi, J., Yoshida, Y., Nakazawa, T., Ooneda, G. 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 (1986).
- Clark, W. M., Lessov, N. S., Dixon, M. P., Eckenstein, F. Monofilament intraluminal middle cerebral artery occlusion in the mouse. Neurol Res. 19, 641-648 (1997).
- Fujii, M. Strain-related differences in susceptibility to transient forebrain ischemia in SV-129 and C57black/6 mice. . Stroke; a journal of cerebral circulation. 28, 1805-1810 (1997).
- Bederson, J. B. Evaluation of 2,3,5-triphenyltetrazolium chloride as a stain for detection and quantification of experimental cerebral infarction in rats. Stroke; a journal of cerebral circulation. 17, 1304-1308 (1986).
- Qi, X., Hosoi, T., Okuma, Y., Kaneko, M., Nomura, Y. Sodium 4-phenylbutyrate protects against cerebral ischemic injury. Mol Pharmacol. 66, 899-908 (2004).
- Barone, F. C., Knudsen, D. J., Nelson, A. H., Feuerstein, G. Z., Willette, R. N. Mouse strain differences in susceptibility to cerebral ischemia are related to cerebral vascular anatomy. J Cereb Blood Flow Metab. 13, 683-692 (1993).
- Tureyen, K., Vemuganti, R., Sailor, K. A., Dempsey, R. J. Ideal suture diameter is critical for consistent middle cerebral artery occlusion in mice. Neurosurgery. 56, 196-200 (2005).
- Barber, P. A., Hoyte, L., Colbourne, F., Buchan, A. M. Temperature-regulated model of focal ischemia in the mouse: a study with histopathological and behavioral outcomes. Stroke; a journal of cerebral circulation. 35, 1720-1725 (2004).
- Kapinya, K. J., Prass, K., Dirnagl, U. Isoflurane induced prolonged protection against cerebral ischemia in mice: a redox sensitive mechanism?. Neuroreport. 13, 1431-1435 (2002).
Tags
Intraluminal Middle Cerebral Artery OcclusionMCAO ModelIschemic StrokeLaser Doppler Flowmetry GuidanceMiceThird Leading Cause Of DeathLeading Cause Of DisabilityCostUnited StatesRatModificationMolecular MechanismsBrain InjuryTherapeutic ModalitiesNoninvasive MethodTransgenic StrainsKnockout StrainsFocal Cerebral IschemiaIntraluminal SutureInternal Carotid ArteryBase Of The MCASpontaneous ReperfusionPermanent RetentionVideo DemonstrationDrawbacksPitfalls235-triphenyltetrazolium Chloride (TTC)Cerebral Infarction
Citazione di questo articolo
Ansari, S., Azari, H., McConnell, D. J., Afzal, A., Mocco, J. Intraluminal Middle Cerebral Artery Occlusion (MCAO) Model for Ischemic Stroke with Laser Doppler Flowmetry Guidance in Mice. J. Vis. Exp. (51), e2879, doi:10.3791/2879 (2011).