PUBLICATION
Genetically Encoded Fluorescent Probe for Imaging Apoptosis in vivo with Spontaneous GFP Complementation
- Authors
- Nasu, Y., Asaoka, Y., Namae, M., Nishina, H., Yoshimura, H., Ozawa, T.
- ID
- ZDB-PUB-151125-8
- Date
- 2016
- Source
- Analytical chemistry 88(1): 838-44 (Journal)
- Registered Authors
- Keywords
- none
- MeSH Terms
-
- Animals
- Apoptosis/genetics*
- Cell Survival
- Fluorescent Dyes/metabolism*
- Genetic Complementation Test*
- Green Fluorescent Proteins/genetics*
- Green Fluorescent Proteins/metabolism*
- HeLa Cells
- Humans
- Mitochondria/metabolism
- Mitochondrial Proteins/metabolism
- Tumor Cells, Cultured
- Zebrafish
- PubMed
- 26597767 Full text @ Anal. Chem.
Citation
Nasu, Y., Asaoka, Y., Namae, M., Nishina, H., Yoshimura, H., Ozawa, T. (2016) Genetically Encoded Fluorescent Probe for Imaging Apoptosis in vivo with Spontaneous GFP Complementation. Analytical chemistry. 88(1):838-44.
Abstract
Apoptosis plays a pivotal role in development and tissue homeostasis in multicellular organisms. Dysfunction of apoptosis is involved in many fatal diseases such as cancer. Visualization of apoptosis in living animals is necessary to understand the mechanism of apoptosis-related diseases. Here, we describe a genetically encoded fluorescent probe for imaging apoptosis in living multicellular organisms, based on spontaneous complementation of two fragments of a green fluorescent protein (GFP) variant (GFP OPT). The probe is designed for detection of mitochondria-mediated apoptosis during which a mitochondrial protein of Smac is released into cytosol. The Smac is connected with a carboxy-terminal fragment of GFP OPT (GFP11), whereas the remainder of GFP OPT (GFP1-10) is located in the cytosol. Under an apoptotic condition, the Smac is released from mitochondria into cytosol, allowing complementation of the GFP-OPT fragments and the emission of fluorescence. Live-cell imaging demonstrates that the probe enables detection of apoptosis in living cells with a high signal-to-background ratio. We applied the probe to living zebrafish, in which apoptotic cells were visualized with fluorescence. The technique provides a useful tool for the study of apoptosis in living animals, facilitating elucidation of the mechanisms of apoptosis-related diseases.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping