title: Apical localization of sodium-dependent glucose transporter SGLT1 is maintained by cholesterol and microtubules.
abstract: A GFP-labeled sodium-dependent glucose transporter SGLT1 (SGLT-GFP) was transfected into MDCK cells. SGLT-GFP was localized at the apical membrane in confluent cells. When cellular cholesterol was depleted by methyl-beta-cyclodextrin (MbetaCD) treatment, the localization of SGLT-GFP gradually switched from apical to whole plasma membrane. Time-lapse microscopy revealed that the effect of MbetaCD appeared within 30 min, and that the transition of SGLT-GFP to the whole plasma membrane was completed within 2 hr after the administration. Immunofluorescence microscopy revealed that the tight junction framework remained steady during this process. The effect of MbetaCD on SGLT-GFP localization was counter-balanced by the addition of cholesterol into the culture medium. Disruption of microtubules by colcemid also perturbed SGLT-GFP localization. SGLT-GFP localized to the whole plasma membrane by colcemid treatment, and apical localization was restored within 1 hr after -removal of colcemid. Inhibition of protein synthesis by cycloheximide had no effect on the transition of SGLT-GFP induced by the MbetaCD or colcemid. These results indicated that the apical localization of SGLT-GFP is maintained by cellular cholesterol and microtubules, possibly with an apical recycling machinery.
authors.affiliation: Department of Anatomy and Cell Biology, Gunma University Graduate School of Medicine, Maebashi, Gunma 371-8511, Japan.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Suzuki
authors.forename: Toshiyuki
authors.initials: T
authors.lastname: Matsuzaki
authors.forename: Haruo
authors.initials: H
authors.lastname: Hagiwara
authors.forename: Takeo
authors.initials: T
authors.lastname: Aoki
authors.forename: Yukiko
authors.initials: Y
authors.lastname: Tajika-Takahashi
authors.forename: Kuniaki
authors.initials: K
authors.lastname: Takata