| An ex vivo Approach to Assess Mitochondrial ROS by Flow Cytometry in AAV-tagged Astrocytes in Adult Mice
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Author:
Date:
2020-03-20
[Abstract] Mitochondrial reactive oxygen species (mROS) are naturally produced signalling molecules extremely relevant for understanding both health- and disease-associated biological processes. The study of mROS in the brain is currently underway to decipher their physiopathological roles and contributions in neurological diseases. Recent advances in this field have highlighted the importance of studying mROS signalling and redox biology at the cellular level. Neurons are especially sensitive to the harmful effects of excess mROS while astrocytic mROS have been shown to play a relevant physiological role in cerebral homeostasis and behaviour. However, given the complexity of the brain, investigating mROS formation in a specific cell-type in adult animals is methodologically challenging. Here we ...
[摘要] [摘要] 线粒体活性氧（mROS ）是天然产生的信号分子，与理解健康和疾病相关的生物过程极为相关。目前正在研究大脑中的mROS ，以了解其在神经疾病中的生理病理作用和贡献。该领域的最新进展凸显了在细胞水平研究mROS 信号传导和氧化还原生物学的重要性。神经元对过量的mROS 的有害作用特别敏感，而星形细胞的mROS对 已经显示出在脑稳态和行为中起相关的生理作用。但是，鉴于大脑的复杂性，研究成年动物中特定细胞类型中mROS的形成在方法上具有挑战性。在这里，我们提出了一种方法来具体评估星形胶质细胞中的mROS 丰度，该方法结合了i ）一种靶向策略，该策略基于在星形胶质细胞（胶质纤维酸性蛋白或GFAP）下表达绿色荧光蛋白（GFP）的腺相关病毒（AAV）载体的使用）启动子，以及ii）使用商业探针通过流式细胞术测量mROS 的强大且广泛扩展的方案。这项工作的意义在于，它允许通过易于使用的技术来选择性研究星形细胞mROS的丰度。
[背景 ] 氧化损伤与许多疾病的病因有关，包括神经退行性疾病，因为大脑是氧化应激以及与年龄相关的变化异常脆弱的组织（Cobley 等，2018; Mattson和Arumugam ...
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| Bone Marrow-derived Endothelial Progenitor Cell Intercellular Adhesion Assay
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Author:
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2016-08-20
[Abstract] Cell-cell adhesion ensures tight contacts between neighbouring cells, which is necessary for cell segregation, as well as for the morphological and functional differentiation of different tissues. Evidently there are cell-cell recognition systems that make cells of the same type preferentially adherent to one another. Homotypic cell adhesion is particularly important in mediating a range of physiological processes such as cell survival, migration and remodeling of vessels. Thus in the present study we selected two populations of endothelial progenitor cells which are from the same donor to investigate the possible effect of a small molecule compound Icariin on homotypic cell adhesion. Many angiogenic factors can destabilize the organization of intercellular junctions, causing endothelial ...
[摘要] 细胞粘附保证了相邻细胞之间的紧密接触，这对于细胞分离以及不同组织的形态和功能分化是必要的。显然，存在使相同类型的细胞优先彼此粘附的细胞 - 细胞识别系统。同型细胞粘附在介导一系列生理过程例如细胞存活，迁移和血管重塑中特别重要。因此，在本研究中，我们选择两个来自相同供体的内皮祖细胞群体，以研究小分子化合物Icariin对同型细胞粘附的可能影响。许多血管生成因子可以破坏细胞间连接的组织，导致内皮屏障开放。在本研究中，我们观察到Icariin治疗减少EPC中VE-钙粘蛋白表达的水平，表明细胞 - 细胞粘附减少 - 证明Icariin的促血管生成作用。总之，所观察到的EPCs的同型粘附的丧失可能有助于由Icariin施加的增强的血管生成作用。
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| Assessment of Human Dendritic Cell Antigen Uptake by Flow Cytometry
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Author:
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2013-11-20
[Abstract] Antigen uptake by dendritic cells is the first key step towards induction of antigen-specific T-cell responses. This flow cytometry-based protocol describes the analysis of dendritic cell uptake of soluble antigens through two different mechanisms: non-specific macropinocytosis (using Lucifer Yelloy CH), and receptor-mediated endocytosis (using DQTM Ovalbumin). The protocol is generated based on data presented in Olivar et al. (2013).
[摘要] 树突状细胞的抗原摄取是诱导抗原特异性T细胞应答的第一个关键步骤。 这种基于流式细胞术的方案描述了通过两种不同的机制分析树突状细胞摄取可溶性抗原：非特异性大细胞增多症（使用Lucifer Yelloy CH）和受体介导的内吞作用（使用DQ TM 卵清蛋白） 。 该方案是基于Olivar等人（2013）中提供的数据产生的。
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