title: Aquaporin-2 is retrieved to the apical storage compartment via early endosomes and phosphatidylinositol 3-kinase-dependent pathway.
abstract: Aquaporin-2 (AQP2) is one of the water-channel proteins expressed in principal cells of kidney collecting ducts, where it is stored in the intracellular compartment. Previous studies have demonstrated that AQP2 vesicles constitute a distinct intracellular compartment partially overlapping with early endosomes. In this report, we performed in vitro experiments using the renal epithelial cell line, Madin-Darby canine kidney (MDCK) cells, stably expressing AQP2 (MDCK-hAQP2). In nonpolarized cells, AQP2 vesicles were scattered in the cytoplasm and did not colocalize with Golgi 58K or TGN38. Small portions of AQP2 vesicles were positive for the lysosome marker cathepsin D. An early endosome antigen (EEA1) localized around AQP2 vesicles in close proximity, suggesting involvement of the endosomal system in the trafficking of AQP2. AQP2 vesicles are distinct from other recycling molecules, such as glucose transporter 4 (GLUT4) and endocytosed transferrin. In polarized MDCK-hAQP2 cells, AQP2 vesicles were localized in the subapical recycling compartment and distinct from the Golgi apparatus, trans-Golgi network, lysosome, and early endosome in the nonstimulated state. When the cells were treated with forskolin, translocation of AQP2 to the apical membrane was observed. Washout of forskolin induced retrieval of AQP2 into the cytoplasm, and AQP2 was transiently colocalized with EEA1-positive endosomes. Then, AQP2 moved from EEA1-positive endosomes to the subapical AQP2-storage compartment, which is sensitive to wortmannin and LY294002. These results suggest that AQP2 resides in a recycling compartment at the apical side in polarized MDCK-hAQP2 cells, and its retrieval uses the apical endosomal system and the phosphatidylinositol 3-kinase-dependent pathway.
authors.affiliation: Department of Anatomy and Cell Biology, Gunma University Graduate School of Medicine, 3-39-22 Showa-machi, Maebashi, Gunma 371-8511, Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Tajika
authors.forename: Toshiyuki
authors.initials: T
authors.lastname: Matsuzaki
authors.forename: Takeshi
authors.initials: T
authors.lastname: Suzuki
authors.forename: Takeo
authors.initials: T
authors.lastname: Aoki
authors.forename: Haruo
authors.initials: H
authors.lastname: Hagiwara
authors.forename: Michio
authors.initials: M
authors.lastname: Kuwahara
authors.forename: Sei
authors.initials: S
authors.lastname: Sasaki
authors.forename: Kuniaki
authors.initials: K
authors.lastname: Takata
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies; D051399:Aquaporin 2; D020346:Aquaporins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D002451:Cell Compartmentation / Q000502:physiology*; D002460:Cell Line; D016764:Cell Polarity / Q000502:physiology; D011992:Endosomes / Q000378:metabolism*; D051275:Glucose Transporter Type 4; D006801:Humans; D007668:Kidney / Q000166:cytology; D008969:Molecular Sequence Data; D009004:Monosaccharide Transport Proteins / Q000378:metabolism; D009124:Muscle Proteins; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism*; D021381:Protein Transport / Q000502:physiology; D011990:Receptors, Transferrin / Q000378:metabolism; D014162:Transfection
chemical_list: C496023:AQP2 protein, human; D000906:Antibodies; D051399:Aquaporin 2; D020346:Aquaporins; D051275:Glucose Transporter Type 4; D009004:Monosaccharide Transport Proteins; D009124:Muscle Proteins; D011990:Receptors, Transferrin; C495692:SLC2A4 protein, human; D019869:Phosphatidylinositol 3-Kinases