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Updated: May 28, 2026
Live-cell Imaging of Migrating Cells Expressing Fluorescently-tagged Proteins in a Three-dimensional Matrix
Published on: December 22, 2011
Video-rate bioluminescence imaging of matrix metalloproteinase-2 secreted from a migrating cell
Takahiro Suzuki1, Chihiro Kondo, Takao Kanamori
1Department of Biochemistry, School of Dentistry, Aichi-Gakuin University, Nagoya, Japan. tsuzuki@dpc.agu.ac.jp
Background:
Matrix metalloproteinase-2 (MMP-2) plays an important role in cancer progression and metastasis. MMP-2 is secreted as a pro-enzyme, which is activated by the membrane-bound proteins, and the polarized distribution of secretory and the membrane-associated MMP-2 has been investigated. However, the real-time visualizations of both MMP-2 secretion from the front edge of a migration cell and its distribution on the cell surface have not been reported.
Methodology/Principal Findings:
The method of video-rate bioluminescence imaging was applied to visualize exocytosis of MMP-2 from a living cell using Gaussia luciferase (GLase) as a reporter. The luminescence signals of GLase were detected by a high speed electron-multiplying charge-coupled device camera (EM-CCD camera) with a time resolution within 500 ms per image. The fusion protein of MMP-2 to GLase was expressed in a HeLa cell and exocytosis of MMP-2 was detected in a few seconds along the leading edge of a migrating HeLa cell. The membrane-associated MMP-2 was observed at the specific sites on the bottom side of the cells, suggesting that the sites of MMP-2 secretion are different from that of MMP-2 binding.
Conclusions:
We were the first to successfully demonstrate secretory dynamics of MMP-2 and the specific sites for polarized distribution of MMP-2 on the cell surface. The video-rate bioluminescence imaging using GLase is a useful method to investigate distribution and dynamics of secreted proteins on the whole surface of polarized cells in real time.
Insights
Researchers visualized the secretion dynamics of matrix metalloproteinase-2 (MMP-2) in real-time using bioluminescence imaging. This method revealed distinct sites for MMP-2 secretion and cell surface binding in migrating cells.
Area of Science:
- Cell Biology
- Biochemistry
- Cancer Research
Background:
- Matrix metalloproteinase-2 (MMP-2) is crucial for cancer progression and metastasis.
- Investigated the polarized distribution of secretory and membrane-associated MMP-2.
- Real-time visualization of MMP-2 secretion and cell surface distribution remained unaddressed.
Purpose of the Study:
- To visualize real-time MMP-2 secretion from migrating cells.
- To investigate the distribution of MMP-2 on the cell surface.
- To develop a novel imaging technique for secreted proteins.
Main Methods:
- Employed video-rate bioluminescence imaging with Gaussia luciferase (GLase) reporter.
- Utilized a high-speed EM-CCD camera for rapid signal detection (within 500 ms/image).
- Expressed MMP-2 fused to GLase in HeLa cells.
Main Results:
- Successfully detected MMP-2 exocytosis within seconds along the leading edge of migrating HeLa cells.
- Observed membrane-associated MMP-2 at specific sites on the cell's bottom surface.
- Demonstrated distinct locations for MMP-2 secretion and binding.
Conclusions:
- First successful demonstration of MMP-2 secretory dynamics and polarized cell surface distribution.
- Video-rate bioluminescence imaging with GLase is effective for real-time analysis of secreted proteins.
- Provides insights into the spatial regulation of MMP-2 in cell migration.
