Low-affinity transport of FITC-albumin in alveolar type II epithelial cell line RLE-6TN.
FITC-albumin uptake by cultured alveolar type II epithelial cells, RLE-6TN, is mediated by high- and low-affinity transport systems. In this study, characteristics of the low-affinity transport system were evaluated. The uptake of FITC-albumin was time and temperature dependent and was inhibited by metabolic inhibitors and bafilomycin A1. Confocal laser scanning microscopic analysis showed punctate localization of the fluorescence in the cells, which was partly localized in lysosomes. FITC-albumin taken up by the cells gradually degraded over time, as shown by fluoroimage analyzer after SDS-PAGE. The uptake of FITC-albumin by RLE-6TN cells was not inhibited by caveolae-mediated endocytosis inhibitors such as nystatin, but was inhibited by clathrin-mediated endocytosis inhibitors such as phenylarsine oxide. The uptake was also inhibited by potassium depletion and hypertonicity, conditions known to inhibit clathrin-mediated endocytosis. In addition, macropinocytosis inhibitors such as 5-(N-ethyl-N-isopropyl) amiloride inhibited the uptake. These results indicate that the low-affinity transport of FITC-albumin in RLE-6TN cells is at least in part mediated by clathrin-mediated endocytosis, but not by caveolae-mediated endocytosis. Possible involvement of macropinocytosis was also suggested.
Tagawa
Maki
M
Department of Pharmaceutics and Therapeutics, Graduate School of Biomedical Sciences, Hiroshima University, Hiroshima, Japan.
Yumoto
Ryoko
R
Oda
Keisuke
K
Nagai
Junya
J
Takano
Mikihisa
M
D019297:2,4-Dinitrophenol / Q000494:pharmacology; D000818:Animals; D001152:Arsenicals / Q000494:pharmacology; D001692:Biological Transport / Q000187:drug effects; D021941:Caveolae / Q000502:physiology; D002460:Cell Line; D002746:Chlorpromazine / Q000494:pharmacology; D002966:Clathrin / Q000502:physiology; D003847:Deoxyglucose / Q000494:pharmacology; D004705:Endocytosis / Q000502:physiology; D004847:Epithelial Cells / Q000378:metabolism*; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives* / Q000378:metabolism; D008247:Lysosomes / Q000378:metabolism; D018942:Macrolides / Q000494:pharmacology; D018613:Microscopy, Confocal; D011650:Pulmonary Alveoli / Q000166:cytology*; D051381:Rats; D012709:Serum Albumin / Q000378:metabolism*; D019810:Sodium Azide / Q000494:pharmacology
D001152:Arsenicals; D002966:Clathrin; C518288:FITC-albumin; D018942:Macrolides; D012709:Serum Albumin; C029341:oxophenylarsine; C040929:bafilomycin A1; D019810:Sodium Azide; D003847:Deoxyglucose; D016650:Fluorescein-5-isothiocyanate; D019297:2,4-Dinitrophenol; D002746:Chlorpromazine