Protocol for imaging and quantifying single-cell CDK activity levels during early mouse embryonic development by...

Area of Science:

• Developmental Biology
• Cell Biology
• Molecular Biology

Background:

• Quantifying protein activity in single cells during early embryonic development is challenging.
• Cyclin-dependent kinases (CDKs) are crucial regulators of the cell cycle in pre-implantation embryos.

Purpose of the Study:

• To provide a detailed protocol for measuring cyclin-dependent kinase (CDK) activity in single cells of mouse pre-implantation embryos.
• To enable researchers to track CDK activity dynamics throughout early embryonic development.

Main Methods:

• Utilizing fluorescent-based reporters for CDK activity.
• Employing in vitro culture strategies for live imaging of mouse embryos.
• Implementing live fluorescent confocal microscopy for image acquisition.
• Developing computational workflows for cell tracking and data analysis.

Main Results:

• The protocol allows for the quantification of CDK activity at the single-cell level.
• Efficient live imaging and cell tracking strategies are described.
• A comprehensive image analysis workflow for processing and visualization is detailed.

Conclusions:

• This approach simplifies the complex task of measuring CDK activity in developing mouse embryos.
• The protocol facilitates the study of cell cycle regulation during pre-implantation development.
• This method provides valuable insights into the role of CDKs in early embryogenesis.