Transcytosis of polymeric immunoglobulin a in polarized Madin-Darby canine kidney cells.
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.
Oztan
Asli
A
Department of Medicine, University of Pittsburgh, Pittsburgh, PA, USA.
Rondanino
Christine
C
Apodaca
Gerard
G
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
D007070:Immunoglobulin A; D007457:Iodine Radioisotopes; D019056:Receptors, Polymeric Immunoglobulin; D014238:Trichloroacetic Acid