Genetically Encoded Fluorescent Probe for Imaging Apoptosis in Vivo with Spontaneous GFP Complementation
Yusuke Nasu1, Yoichi Asaoka2, Misako Namae2
1Department of Chemistry, School of Science, The University of Tokyo , 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
Analytical Chemistry
|November 25, 2015
Summary
We developed a novel fluorescent probe for imaging apoptosis in living organisms. This tool allows real-time visualization of programmed cell death, aiding disease mechanism research.
Area of Science:
- Cell Biology
- Molecular Biology
- Biotechnology
Background:
- Apoptosis is crucial for development and tissue homeostasis.
- Dysfunctional apoptosis is linked to diseases like cancer.
- Visualizing apoptosis in vivo is essential for understanding disease mechanisms.
Purpose of the Study:
- To develop a genetically encoded fluorescent probe for imaging apoptosis in living multicellular organisms.
- To enable visualization of mitochondria-mediated apoptosis.
- To provide a tool for studying apoptosis-related diseases in vivo.
Main Methods:
- Utilized a genetically encoded fluorescent probe based on split green fluorescent protein (GFP) complementation (GFP OPT).
- Designed the probe for detecting Smac release from mitochondria during apoptosis.
- Applied live-cell imaging and in vivo imaging in zebrafish.
Main Results:
- The probe enables detection of apoptosis in living cells with a high signal-to-background ratio.
- Successfully visualized apoptotic cells in living zebrafish using the developed probe.
- Demonstrated the probe's efficacy in live-cell and in vivo imaging.
Conclusions:
- The genetically encoded fluorescent probe is effective for imaging apoptosis in living multicellular organisms.
- This technique offers a valuable tool for studying apoptosis and related diseases in vivo.
- Facilitates elucidation of apoptosis mechanisms in living animals.
