Live images of GLUT4 protein trafficking in mouse primary hypothalamic neurons using deconvolution microscopy

Chun Austin Changou, Szu Yi Chou, Reni Ajoy

Research output: Contribution to journalArticlepeer-review

5 Citations (Scopus)


Type 2 diabetes mellitus (T2DM) is a global health crisis which is characterized by insulin signaling impairment and chronic inflammation in peripheral tissues. The hypothalamus in the central nervous system (CNS) is the control center for energy and insulin signal response regulation. Chronic inflammation in peripheral tissues and imbalances of certain chemokines (such as CCL5, TNFa, and IL-6) contribute to diabetes and obesity. However, the functional mechanism(s) connecting chemokines and hypothalamic insulin signal regulation still remain unclear. In vitro primary neuron culture models are convenient and simple models which can be used to investigate insulin signal regulation in hypothalamic neurons. In this study, we introduced exogeneous GLUT4 protein conjugated with GFP (GFP-GLUT4) into primary hypothalamic neurons to track GLUT4 membrane translocation upon insulin stimulation. Time-lapse images of GFP-GLUT4 protein trafficking were recorded by deconvolution microscopy, which allowed users to generate high-speed, high-resolution images without damaging the neurons significantly while conducting the experiment. The contribution of CCR5 in insulin regulated GLUT4 translocation was observed in CCR5 deficient hypothalamic neurons, which were isolated and cultured from CCR5 knockout mice. Our results demonstrated that the GLUT4 membrane translocation efficiency was reduced in CCR5 deficient hypothalamic neurons after insulin stimulation.

Original languageEnglish
Article numbere56409
JournalJournal of Visualized Experiments
Issue number130
Publication statusPublished - Dec 7 2017


  • Deconvolution microscopy
  • GLUT4
  • Issue 130
  • Liposome based transfection
  • Live image recording
  • Neurobiology
  • Primary mouse hypothalamic neuron culture

ASJC Scopus subject areas

  • General Neuroscience
  • General Chemical Engineering
  • General Biochemistry,Genetics and Molecular Biology
  • General Immunology and Microbiology


Dive into the research topics of 'Live images of GLUT4 protein trafficking in mouse primary hypothalamic neurons using deconvolution microscopy'. Together they form a unique fingerprint.

Cite this