Methodologies for Following EMT In Vivo at Single Cell Resolution

Area of Science:

• Developmental Biology
• Cell Biology
• Genomics

Background:

• Epithelial-mesenchymal transition (EMT) is a fundamental developmental process occurring in metazoan embryos during mesoderm differentiation.
• Studying EMT in model organisms is challenging due to the small proportion of transitioning cells and difficulties in isolating them.
• Existing methods often struggle to isolate molecular details of EMT from the broader cellular environment.

Purpose of the Study:

• To develop and apply a method for analyzing gene expression specifically in cells undergoing EMT.
• To overcome the limitations of studying minority cell populations in complex embryonic systems.
• To investigate the molecular mechanisms of EMT in sea urchin skeletogenic cells.

Main Methods:

• Utilized single-cell RNA sequencing (scRNA-seq) to profile gene expression.
• Focused analysis on sea urchin skeletogenic cells, a specific population undergoing EMT.
• Enabled in vivo gene expression analysis without confounding signals from other cell types.

Main Results:

• Successfully applied scRNA-seq to isolate and analyze gene expression in sea urchin skeletogenic cells during EMT.
• Demonstrated the feasibility of studying gene trajectories in a minority cell population.
• Provided a method to examine genes expressed before, during, and after the EMT process.

Conclusions:

• Single-cell RNA sequencing offers a powerful tool to study EMT in specific cell populations within developing embryos.
• This approach enhances the ability to uncover the molecular underpinnings of EMT, even in rare cell types.
• The method provides a new avenue for detailed mechanistic studies of EMT in the sea urchin model system.