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Studying the Stoichiometry of Epidermal Growth Factor Receptor in Intact Cells using Correlative Microscopy
09:16

Studying the Stoichiometry of Epidermal Growth Factor Receptor in Intact Cells using Correlative Microscopy

Published on: September 11, 2015

8.7K

Label-free imaging of epidermal growth factor receptor-induced response in single living cells

Zanying Peng1, Jin Lu2, Ling Zhang1

  • 1Department of Chemistry, Key Laboratory of Bioorganic Phosphorus Chemistry & Chemical Biology, Tsinghua University, Beijing 100084, China. jhli@mail.tsinghua.edu.cn.

The Analyst
|October 4, 2018
PubMed

Insights

This study visualizes epidermal growth factor receptor (EGFR) activation dynamics in real-time using SPR microscopy. It reveals distinct signaling phases and inhibitor effects on EGFR pathways in A431 cells.

Area of Science:

  • Cell biology
  • Biophysics
  • Cancer research

Background:

  • Epidermal growth factor receptor (EGFR) signaling is vital for cell functions and implicated in numerous cancers.
  • Understanding the spatial and temporal dynamics of EGFR signaling cascades remains a significant challenge.

Purpose of the Study:

  • To visualize real-time EGFR activation and signaling dynamics in response to epidermal growth factor (EGF) in single A431 cells.
  • To investigate the spatial and temporal characteristics of EGFR signaling using advanced microscopy techniques.

Main Methods:

  • Utilized surface plasmon resonance (SPR) microscopy for real-time imaging of EGFR activation.
  • Employed inhibitor analysis with AG1478 and cytochalasin B to probe signaling pathways.

Main Results:

  • Observed a two-phase SPR response pattern within 30 minutes of EGF treatment.
  • Identified a positive SPR signal linked to mass redistribution within 600 seconds, followed by a negative signal due to cell morphology changes.
  • AG1478 inhibited the response across the entire cell, while cytochalasin B specifically affected the cell edge region.

Conclusions:

  • SPR microscopy provides a powerful tool for real-time monitoring of EGFR signaling dynamics.
  • EGFR activation involves distinct temporal phases and spatial redistribution of cellular components.
  • Inhibitor studies highlight the differential roles of specific pathways in EGFR signaling across the cell.

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