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Fluorescence-based visualization of autophagic activity predicts mouse embryo viability
Satoshi Tsukamoto1, Taichi Hara2, Atsushi Yamamoto3
1Laboratory Animal and Genome Sciences Section, National Institute of Radiological Sciences, 4-9-1 Anagawa, Inage-ku, Chiba 263-8555, Japan.
Scientific Reports
|April 1, 2014
Summary
Embryonic autophagic activity, a key cellular process, declines with maternal age. Measuring this activity using GFP-LC3 fluorescence can predict embryo quality in assisted reproduction.
Area of Science:
- Reproductive biology
- Cellular biology
- Molecular mechanisms of development
Background:
- Embryo quality is crucial for assisted reproductive technologies, but morphological assessment has limitations.
- Autophagy, a cellular degradation process, is vital for embryonic development.
- Understanding molecular mechanisms of embryo quality control is essential.
Purpose of the Study:
- To develop a method for visualizing and quantifying autophagic activity in live mouse embryos.
- To investigate the relationship between maternal age, embryonic autophagic activity, and embryo viability.
Main Methods:
- Development of a simple fluorescence-based assay using GFP-LC3 microinjection to monitor autophagy.
- Imaging and quantification of GFP-LC3 fluorescence intensity in live mouse embryos.
- Correlation analysis of autophagic activity with maternal age and embryo developmental viability.
Main Results:
- A novel method for visualizing live embryonic autophagic activity was established.
- Embryonic autophagic activity was found to decrease with increasing maternal age.
- Reduced autophagic activity correlated with diminished embryo viability.
Conclusions:
- Embryonic autophagic activity, visualized via GFP-LC3, is a potential indicator of embryo quality.
- The decline in autophagic activity with maternal age may involve lysosomal hydrolase function.
- This method offers a new avenue for assessing embryo viability in reproductive technologies.
