Transcellular iodide transport and iodination on the apical plasma membrane by monolayer porcine thyroid cells cultured on collagen-coated filters.
Isolated porcine thyroid follicular cells were cultured on a collagen-coated Millipore filter to form a monolayer. The monolayer could translocate 125I added in the medium beneath the filter (basal medium) into the medium above the monolayer (apical medium) and form an iodide concentration gradient of several-fold. Transcellular iodide pump activity was observed when the cells were cultured with TSH in the basal medium. In the absence of TSH, the translocation of iodide was very slow. The concentration of TSH required to activate the iodide pump was 0.1-0.3 mU/ml. Addition of ClO4- to the basal medium inhibited transcellular transport, whilst addition of ClO4- to the apical medium was much less effective. Constituents labelled with 125I in the apical medium were analysed. The amount of protein-bound 125I measured by acid precipitation was 3-8% of the total radioactivity. The residual radioactivity was found to be iodide ion by paper chromatography. Further analysis by sodium dodecylsulphate-polyacrylamide gel electrophoresis revealed that most of the 125I-labelled protein was at the position of bovine serum albumin which had been added to the culture medium. The monolayer culture of cells on collagen-coated filter would be a useful experimental system for analysing thyroid cell functions for which the cell polarity is essential.
Central Laboratory for Clinical Investigation, Miyazaki Medical College Hospital, Japan.
Y
Y
Nakamura
T
T
Kotani
S
S
Ohtaki
D000818:Animals; D001692:Biological Transport; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D003094:Collagen / Q000378:metabolism; D007454:Iodides / Q000378:metabolism*; D008851:Micropore Filters; D051381:Rats; D013552:Swine; D013961:Thyroid Gland / Q000166:cytology / Q000378:metabolism*; D013972:Thyrotropin / Q000378:metabolism; D013997:Time Factors
D007454:Iodides; D013972:Thyrotropin; D003094:Collagen