Selective radioiodination of the apical and luminal cell surfaces: in vitro and in situ experiments on vascular endothelial cells with Iodogen-coated Sephadex.
A gentle and nonexpensive agent for selective radioiodination of the cell surface proteins was obtained by plating aliquots of Iodogen on dried Sephadex beads 50-60 microns in diameter. Iodogen-coated Sephadex inherits Iodogen properties: it is stable and virtually insoluble in water, allowing rapid iodination of the cell surface proteins in the solid phase with 125I-. Iodination is terminated by simply removing the beads. The agent was tested on bovine aortic endothelial cells in culture and on rabbit aortic endothelial cells in situ. Light and electron microscopic studies revealed that during radioiodination, apparently no ultrastructural modifications occurred in the endothelial cells. In addition, experiments with 51Cr (used as an indicator of endothelial cell injury) demonstrated that during iodination the cell integrity was preserved. The technique reported here may be generally applied for selective radioiodination of the apical surface proteins of various cultured cells and of the luminal endothelial surface of large blood vessels.
Ghinea
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N
Institute of Cellular Biology and Pathology, Bucharest, Romania.
Hasu
M
M
Popov
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D
D000818:Animals; D001011:Aorta / Q000378:metabolism* / Q000648:ultrastructure; D001345:Autoradiography; D002417:Cattle; D003911:Dextrans; D004591:Electrophoresis, Polyacrylamide Gel; D004730:Endothelium, Vascular / Q000378:metabolism*; D007202:Indicators and Reagents; D007457:Iodine Radioisotopes; D008297:Male; D011817:Rabbits; D014508:Urea / Q000031:analogs & derivatives
D003911:Dextrans; D007202:Indicators and Reagents; D007457:Iodine Radioisotopes; C012385:1,3,4,6-tetrachloro-3 alpha,6 alpha-diphenylglycoluril; D014508:Urea; C025614:sephadex