Close Association of Aquaporin-2 Internalization with Caveolin-1
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概要
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Aquaporin 2 (AQP2) is a membrane water channel protein that traffics between the intracellular membrane compartment and the plasma membrane in a vasopressin-dependent manner in the renal collecting duct cell to control the amount of water reabsorption. We examined the relation between AQP2 internalization from the plasma membrane and caveolin-1, which is a major protein in membrane microdomain caveolae, in Mardin-Darby canine kidney cells expressing human AQP2 (MDCK-hAQP2 cells). Double-immunofluorescence microscopy showed that AQP2 is colocalized with caveolin-1 in the apical plasma membrane by stimulating the intracellular signaling cascade of vasopressin with forskolin. After washing forskolin, both AQP2 and caveolin-1 were internalized to early endosomes and then separately went back to their individual compartments, which are subapical compartments and the apical membrane, respectively. Double-immunogold electron microscopy in ultrathin cryosections confirmed the colocalization of AQP2 with caveolin-1 at caveolar structures on the apical plasma membrane of forskolin-treated cells and the colocalization within the same intracellular vesicles after washing forskolin. A co-immunoprecipitation experiment showed the close interaction between AQP2 and caveolin-1 in forskolin-treated cells and in cells after washing forskolin. These results suggest that a caveolin-1-dependent and possibly caveolar-dependent pathway is a candidate for AQP2 internalization in MDCK cells.
著者
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Sasaki Sei
Department Of Nephrology Graduate School Of Medicine Tokyo Medical And Dental University
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Aoki Takeo
Department Of Anatomy And Cell Biology Gunma University Graduate School Of Medicine
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Takata Kuniaki
Department Of Anatomy And Cell Biology Gunma University Graduate School Of Medicine
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Hagiwara Haruo
Department Of Anatomy And Cell Biology Gunma University Graduate School Of Medicine
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Kuwahara Michio
Division Of Nephrology Shuwa General Hospital
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Matsuzaki Toshiyuki
Department Of Anatomy And Cell Biology Gunma University Graduate School Of Medicine
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Suzuki Takeshi
Department Of Allergy And Rheumatology Graduate School Of Medicine University Of Tokyo
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Aoki Takeo
Department of Anatomy and Cell Biology, Gunma University Graduate School of Medicine
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