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Quantitative Measurement of GLUT4 Translocation to the Plasma Membrane by Flow Cytometry
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Quantitative Measurement of GLUT4 Translocation to the Plasma Membrane by Flow Cytometry

Published on: November 7, 2010

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Novel dual-color drug screening model for GLUT4 translocation in adipocytes

Yanting Lu1, Xiuli Ma1, Qinghua Kong2

  • 1State Key Laboratory of Phytochemistry and Plant Resources in West China, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming, 650201, China; University of the Chinese Academy of Sciences, Beijing, 100049, China.

Molecular and Cellular Probes
|January 15, 2019
PubMed

Insights

Researchers developed a novel pHluorin-GLUT4-mOrange2 probe and a screening system using 3T3-L1 adipocytes to identify compounds that enhance glucose transporter type 4 (GLUT4) translocation. This system aids in discovering new type 2 diabetes drugs.

Area of Science:

  • Biochemistry
  • Cell Biology
  • Pharmacology

Background:

  • Insulin-responsive glucose transporter type 4 (GLUT4) translocation is critical for glucose uptake in adipose tissue and muscle, maintaining metabolic homeostasis and preventing hyperglycemia.
  • Identifying compounds that enhance GLUT4 translocation is a key strategy for developing new type 2 diabetes therapeutics.

Purpose of the Study:

  • To develop a novel, functional dual-color probe, pHluorin-GLUT4-mOrange2, for monitoring GLUT4 translocation.
  • To establish an efficient screening system based on 3T3-L1 adipocytes for identifying compounds that stimulate GLUT4 translocation.
  • To facilitate high-throughput screening for drug discovery targeting glucose uptake.

Main Methods:

  • Development of a dual-color fluorescent probe (pHluorin-GLUT4-mOrange2) to visualize GLUT4.
  • Construction of a screening system utilizing 3T3-L1 adipocytes engineered with the probe.
  • Application of the system to screen for compounds that promote GLUT4 translocation.

Main Results:

  • Successful development and implementation of the pHluorin-GLUT4-mOrange2 probe and screening system.
  • Identification of several chemical compounds that effectively facilitate GLUT4 translocation in 3T3-L1 adipocytes.
  • Demonstration of the system's efficiency in screening for GLUT4 translocation modulators.

Conclusions:

  • A novel, user-friendly dual-color GLUT4 probe has been developed to monitor translocation in insulin-responsive cells.
  • This system offers an alternative method for high-throughput screening of compounds that regulate GLUT4 translocation and glucose uptake.
  • The probe and system can be utilized to investigate the molecular mechanisms of GLUT4 trafficking, including its approach, docking, and fusion with the plasma membrane.
Keywords:
GLUT4Glucose uptakeScreeningpHluorin

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