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Updated: Sep 30, 2025
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Visualizing Clathrin-mediated Endocytosis of G Protein-coupled Receptors at Single-event Resolution via TIRF Microscopy
Published on: October 20, 2014
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Single molecule localization-based analysis of clathrin-coated pit and caveolar dynamics
Rui Ma1, Martin Štefl1, Gerd Ulrich Nienhaus1,2,3,4
1Institute of Applied Physics (APH), Karlsruhe Institute of Technology (KIT), Wolfgang Gaede-Strasse 1, 76131 Karlsruhe, Germany. uli@uiuc.edu.
Nanoscale Horizons
|March 15, 2022
Summary
Super-resolution microscopy reveals clathrin-coated pits and caveolae assembly dynamics in live cells. Shared accessory proteins appear to orchestrate the development of these crucial plasma membrane structures.
Area of Science:
- Cell biology
- Biophysics
- Microscopy
Background:
- Clathrin-coated pits and caveolae are essential nanoscale plasma membrane invaginations involved in cellular processes.
- Their formation involves complex protein assembly regulated by accessory proteins.
Purpose of the Study:
- To analyze the development of clathrin-coated and caveolar structures with high resolution in live cells.
- To investigate the dynamics and temporal evolution of these membrane coat structures.
Main Methods:
- Utilized super-resolution single-molecule localization microscopy (SMLM) on live cells.
- Developed a sophisticated clustering and data analysis workflow for SMLM image sequences.
- Quantified structure lifetimes and growth dynamics.
Main Results:
- Observed similar temporal development patterns for both clathrin-coated and caveolar structures.
- Identified hotspots on the plasma membrane where coat structures repeatedly form.
- Quantified lifetime distributions and growth dynamics of these membrane invaginations.
Conclusions:
- Key accessory proteins, some shared between clathrin and caveolae pathways, likely orchestrate their temporal development.
- The study provides novel insights into the dynamic assembly of essential cellular membrane structures.
- High-resolution live-cell imaging reveals conserved principles in membrane coat formation.
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