极化的犬肾上皮细胞（Madin-Darby canine kidney cells）中聚合免疫球蛋白A的转胞吞作用
Transcytosis of polymeric immunoglobulin a in polarized Madin-Darby canine kidney cells.
作者信息
Oztan Asli, Rondanino Christine, Apodaca Gerard
机构信息
Department of Medicine, University of Pittsburgh, Pittsburgh, PA, USA.
出版信息
Methods Mol Biol. 2008;440:157-70. doi: 10.1007/978-1-59745-178-9_12.
Abstract
The transcytotic pathway allows for the bidirectional transport of endocytosed solutes, lipids, and proteins between the two membrane domains of polarized epithelial cells while maintaining the functional integrity of the epithelial tissue. A method is described to measure basolateral-to-apical transcytosis of immunoglobulin A (IgA) in polarized Madin-Darby canine kidney (MDCK) cells expressing the polymeric immunoglobulin receptor (pIgR). The cells are grown on porous Transwell filter supports, and radiolabeled (125)I-immunoglobulin A (IgA) is internalized from the basolateral pole of MDCK cells. During a subsequent 2-h chase, the amount of (125)I-IgA that is recycled, degraded, or transcytosed is quantified. This assay can be adapted to follow the postendocytic fate of other (125)I-labeled ligands and proteins.
摘要
转胞吞途径允许内吞的溶质、脂质和蛋白质在极化上皮细胞的两个膜结构域之间进行双向运输，同时维持上皮组织的功能完整性。本文描述了一种方法，用于测量表达多聚免疫球蛋白受体（pIgR）的极化犬肾（MDCK）细胞中免疫球蛋白A（IgA）从基底外侧到顶端的转胞吞作用。细胞生长在多孔Transwell滤器支持物上，放射性标记的（125）I-免疫球蛋白A（IgA）从MDCK细胞的基底外侧极内化。在随后的2小时追踪过程中，对回收、降解或转胞吞的（125）I-IgA量进行定量。该测定法可用于追踪其他（125）I标记的配体和蛋白质的内吞后命运。
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