title: Transcytosis of polymeric immunoglobulin a in polarized Madin-Darby canine kidney cells.
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.
authors: Department of Medicine, University of Pittsburgh, Pittsburgh, PA, USA.
authors: Asli
authors: A
authors: Oztan
authors: Christine
authors: C
authors: Rondanino
authors: Gerard
authors: G
authors: Apodaca
mesh_terms: D000818:Animals; D001681:Biological Assay / Q000379:methods* / Q000706:statistics & numerical data; D002460:Cell Line; D016764:Cell Polarity; D011232:Chemical Precipitation; D003627:Data Interpretation, Statistical; D004285:Dogs; D004705:Endocytosis; D004847:Epithelial Cells / Q000378:metabolism*; D007070:Immunoglobulin A / Q000378:metabolism*; D007457:Iodine Radioisotopes; D007668:Kidney / Q000166:cytology / Q000378:metabolism*; D019056:Receptors, Polymeric Immunoglobulin / Q000235:genetics / Q000378:metabolism; D014162:Transfection; D022161:Transport Vesicles / Q000378:metabolism*; D014238:Trichloroacetic Acid / Q000737:chemistry
chemical_list: D007070:Immunoglobulin A; D007457:Iodine Radioisotopes; D019056:Receptors, Polymeric Immunoglobulin; D014238:Trichloroacetic Acid