| Flow Cytometry of Lung and Bronchoalveolar Lavage Fluid Cells from Mice Challenged with Fluorescent Aspergillus Reporter (FLARE) Conidia
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Author:
Date:
2016-09-20
[Abstract] Aspergillus fumigatus is a ubiquitous fungal pathogen that forms airborne conidia. The process of restricting conidial germination into hyphae by lung leukocytes is critical in determining infectious outcomes. Tracking the outcome of conidia-host cell encounters in vivo is technically challenging and an obstacle to understanding the molecular and cellular basis of antifungal immunity in the lung. Here, we describe a method that utilizes a genetically engineered Aspergillus strain [called FLARE (Jhingran et al., 2012; Espinosa et al., 2014; Heung et al., 2015)] to monitor conidial phagocytosis and killing by leukocytes within the lung environment at single encounter resolution.
[摘要] Aspergillus fumigatus is a ubiquitous fungal pathogen that forms airborne conidia. The process of restricting conidial germination into hyphae by lung leukocytes is critical in determining infectious outcomes. Tracking the outcome of conidia-host cell encounters in vivo is technically challenging and an obstacle to understanding the molecular and cellular basis of antifungal immunity in the lung. Here, we describe a method that utilizes a genetically engineered Aspergillus strain [called FLARE (Jhingran et al., 2012; Espinosa et al., 2014; Heung et al., 2015)] to monitor conidial phagocytosis ...
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| Targeted Occlusion of Individual Pial Vessels of Mouse Cortex
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Author:
Date:
2013-09-05
[Abstract] Targeted photothrombosis is a method to occlude individual arterioles and venules that lie on the surface of the cerebral cortex. It has been used to study collateral flow patterns within the pial vascular network following occlusion of single surface vessels (Schaffer et al., 2006; Blinder et al., 2010; Nguyen et al., 2011), as well as to generate localized ischemic strokes following occlusion of single penetrating vessels (Nishimura et al., 2007; Drew et al., 2010; Shih et al., 2013). The intravascular clot is formed by irradiation of a target vessel with a focused green laser after injection of a circulating photosensitizing agent, Rose Bengal (Watson et al., 1985). We briefly describe modifications of custom-designed and ...
[摘要] 目标光血栓形成是阻塞位于大脑皮质表面上的个体小动脉和小静脉的方法。它已经用于在封闭单表面血管后研究在Pial血管网内的侧支血流模式（Schaffer等人，2006; Blinder等人，2010; Nguyen 2011），以及在单一穿透血管闭塞后产生局部缺血性中风（Nishimura et al。，2007; Drew et al。 ，2010; Shih et al。，2013）。在注射循环的光敏剂，玫瑰红（Watson等人，1985）后，通过用聚焦的绿色激光器照射目标血管形成血管内凝块。我们简要介绍了定制设计和商业双光子成像系统的修改，引入绿色激光用于光血栓形成。我们进一步提供如何闭塞麻醉鼠标的体感皮层内的单个穿透小动脉的说明。
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| Mouse Transient Global Ischemia Two-Vessel Occlusion Model
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Author:
Date:
2012-09-20
[Abstract] Transient global ischemia in rodents induces delayed death of hippocampal CA1 neurons, as well as in some hilar neurons of the dentate gyrus, medium aspiny neurons of the striatum, pyramidal neurons in neocortical layers II, V and VI, and Purkinje neurons of the cerebellum. In contrast to focal ischemia that mimics regional stroke in humans, this model of global ischemia mimics the brain injury that occurs after human cardiac arrest. Early events include caspase activation, cleavage of anti-death Bcl-2 family proteins and large mitochondrial channel activity. Genetically engineered mice provide opportunities for study such as the knock-in mouse expressing a caspase-resistant form of Bcl-xL found to exhibit markedly reduced mitochondrial channel activity and reduced vulnerability to ...
[摘要] 啮齿类动物的瞬时全局性缺血诱导海马CA1神经元的延迟死亡，以及在齿状回的一些肝门神经元，纹状体的中度aspiny神经元，新皮质层II，V和VI中的锥体神经元和小脑的浦肯野神经元。与模拟人类的局部中风的局部缺血相反，这种全局缺血模型模拟了在人类心脏骤停后发生的脑损伤。早期事件包括胱天蛋白酶活化，抗死亡Bcl-2家族蛋白的切割和大线粒体通道活性。基因工程小鼠提供了研究的机会，如敲入小鼠表达的caspase抗性形式的Bcl-xL发现表现出显着减少的线粒体通道活动和减少缺血诱导的神经元死亡的脆弱性1。因此，适应和开发一个简单的协议，以在小鼠中产生瞬时全局缺血是相关的。双血管闭塞模型已被专门开发以在小鼠中提供最佳结果，并且相对于传统上用于大鼠的四脉管闭塞模型提供了若干优点，包括相对容易的程序以及仅一天的手术。然而，应当注意，与其他缺血模型相比，该程序具有更高的发病率以及更高程度的变异性。这两个缺点需要使用更大的动物群，对于许多健康的育种转基因动物是非威慑因子。
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