Apical Localization of Sodium-Dependent Glucose Transporter SGLT1 is Maintained by Cholesterol and Microtubules

نویسندگان

  • Takeshi Suzuki
  • Toshiyuki Matsuzaki
  • Haruo Hagiwara
  • Takeo Aoki
  • Yukiko Tajika-Takahashi
  • Kuniaki Takata
چکیده

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.

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عنوان ژورنال:

دوره 39  شماره 

صفحات  -

تاریخ انتشار 2006