分析免疫突触处的肌动蛋白动力学。
Analyzing Actin Dynamics at the Immunological Synapse.
作者信息
Jankowska Katarzyna I, Burkhardt Janis K
机构信息
Department of Pathology and Laboratory Medicine, Children's Hospital of Philadelphia Research Institute, 3615 Civic Center Blvd, ARC 816D, Philadelphia, PA, 19104, USA.
Perelman School of Medicine of the University of Pennsylvania, Philadelphia, PA, USA.
出版信息
Methods Mol Biol. 2017;1584:7-29. doi: 10.1007/978-1-4939-6881-7_2.
Abstract
T cell signaling is inextricably linked to actin cytoskeletal dynamics at the immunological synapse (IS). This process can be imaged in living T cells expressing GFP actin or fluorescent F-actin binding proteins. Because of its planar nature, the IS provides a unique opportunity to image events as they happen, monitoring changes in actin retrograde flow in T cells interacting with different stimulatory surfaces or after pharmacological treatments. Here, we described the imaging methods and analytical procedures used to measure actin velocity across the IS in T cells spreading on planar stimulatory surfaces.
摘要
T细胞信号传导与免疫突触（IS）处的肌动蛋白细胞骨架动力学有着千丝万缕的联系。这一过程可以在表达绿色荧光蛋白（GFP）肌动蛋白或荧光F-肌动蛋白结合蛋白的活T细胞中成像。由于免疫突触的平面性质，它提供了一个独特的机会来实时成像事件，监测T细胞与不同刺激表面相互作用时或经过药物处理后肌动蛋白逆行流的变化。在这里，我们描述了用于测量在平面刺激表面上铺展的T细胞中免疫突触处肌动蛋白速度的成像方法和分析程序。
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