| Microinjection of Virus into Lumbar Enlargement of Spinal Dorsal Horn in Mice
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Author:
Date:
2016-11-20
[Abstract] In order to explore the role of a specific gene/protein in the specific segment of the spinal cord, the technique of intraspinal injection is particularly used to deliver viral vectors targeting the specific gene/protein. These viral vectors can knockdown or overexpress the specific gene/protein in specific cells (glial cells or neurons). In this protocol, lentivirus containing shRNA for CXCL13 were injected into the dorsal horn of the spinal lumbar enlargement segment (Jiang et al., 2016). This technique allows the study of the role of CXCL13 in the ipsilateral dorsal horn in ...
[摘要] 为了探索特定基因/蛋白质在脊髓特定节段中的作用，脊柱内注射技术特别用于递送靶向特定基因/蛋白质的病毒载体。这些病毒载体可以敲低或过表达特定细胞（神经胶质细胞或神经元）中的特定基因/蛋白。在该协议中，将包含用于CXCL13的shRNA的慢病毒注射到脊柱腰扩大节的背角中（Jiang等人，2016）。这种技术允许研究CXCL13在同侧背角中在神经性疼痛中的作用，而不影响DRG或对侧背角。
[背景] 脊髓背角是一种组织良好和分层的神经元复合体，其将来自身体表面和深部组织的感觉信息传输到大脑。脊髓背角神经元的活动可以通过原始传入，下行纤维以及脊髓神经胶质细胞如星形胶质细胞和小胶质细胞来调节。了解如何脊髓神经元或神经胶质细胞中的特定蛋白质和基因有助于生理和病理性疼痛的机制越来越感兴趣。为了探索特定基因/蛋白质在脊髓中的作用，鞘内注射的技术通常用于递送激动剂，拮抗剂，siRNA或miRNA进入蛛网膜下腔。特定基因的敲除和条件缺失是另一种详尽且昂贵的方法。然而，这些方法的限制是缺乏脊髓节段和细胞类型特异性删除或过表达。这个协议表明脊柱内注射可以传递特定的病毒载体到特定的节段脊髓背角，调查基因的功能区域和细胞类型特定的方式。
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| Phagocytosis Assay of Microglia for Dead Neurons in Primary Rat Brain Cell Cultures
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Author:
Date:
2016-04-20
[Abstract] Clearance of dead brain tissue including the dead neurons through phagocytosis is an endogenous function of microglia in the brain, which is critical for inflammation resolution after ischemic stroke or head trauma. By regulating the function or polarization status of microglia, we may control their phagocytosis efficacy and therefore the cleanup process for the dead brain tissue. We cultured rat cortical neurons and microglia from the same litter of embryos. The cultured neurons are subjected to irradiation for inducing neuronal apoptosis. After labeling with propidium iodide (PI), the dead ...
[摘要] 通过吞噬作用来清除包括死亡神经元在内的死亡脑组织是脑中小胶质细胞的内源性功能，这对缺血性卒中或头部创伤后的炎症分解至关重要。通过调节小胶质细胞的功能或极化状态，我们可以控制其吞噬功效，从而控制死脑组织的清除过程。我们从相同的胚胎培养大鼠皮质神经元和小胶质细胞。培养的神经元经受辐射诱导神经细胞凋亡。用碘化丙啶（PI）标记后，将死亡神经元（DN）暴露于培养的小神经胶质细胞进行吞噬试验。通过计算每个小胶质细胞中的DN数量，我们计算吞噬指数，以量化小胶质细胞对DN的吞噬功效。方案分为4个部分：A）从产前大鼠胚胎培养大鼠皮质神经元，B）将死亡神经元作为吞噬作用靶标，C）培养大鼠脑小胶质细胞，D）定量小神经胶质细胞向死亡神经元的吞噬指数。
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| Mast Cell Dependent Airway Hyperresponsiveness (AHR)
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Author:
Date:
2012-10-05
[Abstract] Asthma is a complex phenotype that involves multiple mechanisms, including adaptive and innate immunity as well as physiological and mechanical changes in the airways. In the models of asthma induced by sensitization and aerosolized allergen exposure in the absence of adjuvant, mast cells facilitate the development of in ammation and airway hyper-responsiveness. This model is useful to analysis of function of mast cells in AHR.
[摘要] 该实验方案的中文版正在准备中．．．
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