Observing GLUT4 Translocation in Live L6 Cells Using Quantum Dots

Sensors

Feng Qu¹  Zubin Chen¹  Xiaoxuan Wang¹  Lingfeng Meng¹  Zhengxing Wu¹ 

[1] Institute of Biophysics and Biochemistry, School of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, China;

关键词: quantum dots;    GLUT4;    confocal microscopy;    translocation;    endocytosis;   

DOI  :  10.3390/s110202077

来源: mdpi

【 摘 要 】

The glucose transporter 4 (GLUT4) plays a key role in maintaining whole body glucose homeostasis. Tracking GLUT4 in space and time can provide new insights for understanding the mechanisms of insulin-regulated GLUT4 translocation. Organic dyes and fluorescent proteins were used in previous studies for investigating the traffic of GLUT4 in skeletal muscle cells and adipocytes. Because of their relative weak fluorescent signal against strong cellular autofluorescence background and their fast photobleaching rate, most studies only focused on particular segments of GLUT4 traffic. In this study, we have developed a new method for observing the translocation of GLUT4 targeted with photostable and bright quantum dots (QDs) in live L6 cells. QDs were targeted to GLUT4myc specifically and internalized with GLUT4myc through receptor-mediated endocytosis. Compared with traditional fluorescence dyes and fluorescent proteins, QDs with high brightness and extremely photostability are suitable for long-term single particle tracking, so individual GLUT4-QD complex can be easily detected and tracked for long periods of time. This newly described method will be a powerful tool for observing the translocation of GLUT4 in live L6 cells.

【 授权许可】

CC BY   
© 2011 by the authors; licensee MDPI, Basel, Switzerland.