横向慢性颅窗口准备启用<em>在体内</em>观察继末节小鼠大脑中动脉闭塞
Özet
远端大脑中动脉分支（缺血）的手术闭塞是实验性卒中研究常用的模式。该原稿描述永久MCAO的基本技术，有横向颅窗口的插入，这提供了在小鼠中纵向活体显微镜的机会相结合。
Abstract
局灶性脑缺血（即缺血性中风）可能会导致重大的脑损伤，导致神经功能严重受损并因此导致运动和认知障碍的主机。它的高患病率带来了严重的健康负担，中风是长期的伤残和死亡全世界1的主要原因之一。神经功能的恢复是，在大多数情况下，只有部分。迄今为止，治疗选择是非常有限的，尤其是由于对溶栓2,3窄时间窗。确定的方法来加速从中风复苏仍尖端医疗的目标;然而，这已经阻碍了机械见解不足到恢复过程。试验行程研究者经常使用局灶性脑缺血啮齿动物模型。除了急性期，中风的研究越来越集中于亚急性期和慢性期脑缺血。大多数研究者中风申请永久或TRAN在小鼠或大鼠大脑中动脉闭塞过性。在患者中，MCA的闭塞是缺血性中风4的最常见的原因之一。除了使用灯丝模型MCA的近端闭塞，远端MCA的外科闭塞可能是在实验性卒中研究5中最常用的模型。远端（到lenticulo-纹动脉的分支）马华支闭塞通常免去了纹状体和主要影响大脑皮层。血管闭塞可以是永久的或暂时的。相对于长期预后病变体积和非常低的死亡率高重复性是这种模式的主要优点。在这里，我们将演示如何进行慢性颅窗口（CW）准备外侧为矢状窦，事后如何手术诱导采用开颅手术方法的窗口下方的远端行程。这种方法可应用于以下通过缺血的急性和慢性的变化序列成像外延照射，共焦激光扫描，和双光子活体显微镜。
Introduction
Stroke is among the principal causes of long-term disability and death worldwide1, coming second after coronary heart disease. In addition, stroke is the primary cause of long-term disability, underscoring its tremendous socioeconomic impact6-8. Beyond acute treatment, investigating new approaches and mechanisms to accelerate and enhance recovery after stroke remains a prime medical goal7.
In the last few decades, data from experimental stroke research has contributed substantially to understanding the complex pathophysiological cascades triggered by ischemia9,10. Excitotoxicity, apoptosis, peri-infarct depolarization, and inflammation have been identified as the most relevant mediators of cell death following focal cerebral ischemia. Moreover, using animal models of cerebral ischemia, important concepts, diagnostic modalities, and therapeutic approaches have been developed and validated (e.g., “penumbra” and thrombolysis)11.
The availability of experimental stroke models, combined with non-invasive imaging modalities (e.g., magnetic resonance imaging (MRI), computed tomography, or laser speckle contrast analysis), enables the researcher to investigate hyperacute and chronic pathophysiological changes induced by the ischemic insult in a longitudinal manner12. Along with studying the spatiotemporal profile of the evolving lesion, changes resembling neuronal plasticity can be investigated and correlated to functional outcomes and histological findings. Within the last few years, further methodological advances have been made using the combination of cerebral ischemia models and in vivo microscopy via cranial windows13. These new techniques allow investigators to analyze the neurovascular unit at the cellular and molecular level, with great analytic power in the acute, subacute, and chronic phases following focal cerebral ischemia14. Moreover, in vivo microscopy imaging of microcirculatory dynamics has revealed novel aspects of cerebral microvasculature function and angioarchitecture, with significant pathophysiological relevance15-17.
In this protocol, we present how to perform a chronic CW preparation lateral to the sagittal sinus and how to surgically induce a distal stroke underneath the window. This mouse model can be applied to sequential imaging of acute, subacute, and chronic changes following focal cerebral ischemia via epi-illuminating, confocal laser scanning, and two-photon intravital microscopy.
Protocol
Representative Results
Discussion
中风是长期的伤残和死亡全世界1的主要原因之一。除了急性期治疗，研究新的方法和机制，加快和提高中风后恢复仍然是一个主要的医疗目标7。试验行程研究者经常使用局灶性脑缺血啮齿动物模型。实际上，模型诱导瞬时或永久MCAO模仿最常见的类型患者4局灶性脑缺血中的一个。除了MCA的近端闭塞，为远侧MCAO手术闭塞灯丝模型可能是在实验性卒中研究5,19中最常?…
Açıklamalar
The authors have nothing to disclose.
Acknowledgements
VP is a participant in the Charité Clinical Scientist Program, funded by the Charité – Universitätsmedizin Berlin and the Berlin Institute of Health. TB is an SNSF PostDoc Mobility fellow. The authors receive grant support from EinsteinStiftung/A-2012-153 to PV.
Materials
| Binocular surgical microscope | Zeiss | Stemi 2000 C | |
| Light source for microscope | Zeiss | CL 6000 LED | |
| Heating pad with rectal probe | FST | 21061-10 | |
| Stereotactic frame | Kopf | Model 930 | |
| Anaethesia system for isoflurane | Draeger | ||
| Isoflurane | Abott | ||
| Dumont forceps #5 | FST | 11251-10 | |
| Dumont forceps #7 | FST | 11271-30 | |
| Bipolar Forceps | Erbe | 20195-501 | |
| Bipolar Forceps | Erbe 20195-022 | ||
| Microdrill | FST 18000-17 | ||
| Needle holder | FST | 12010-14 | |
| 5-0 silk suture | Feuerstein, Suprama | ||
| 7-0 silk suture | Feuerstein,Suprama | ||
| 8-0 silk suture | Feuerstein, Suprama | ||
| Veterinary Recovery Chamber | Peco Services | V1200 | |
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