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In vivo profiling of the endothelium using 'AngioTag' zebrafish
Mayumi F Miller1,2, Leah J Greenspan1, Derek E Gildea3
1Division of Developmental Biology, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD, 20892, USA.
Angiogenesis
|July 4, 2025
Summary
Researchers developed the AngioTag zebrafish to study gene expression in undisturbed endothelial cells. This novel method profiles actively translating messenger RNA in vivo, revealing new endothelial genes and organ-specific signatures.
Area of Science:
- Molecular Biology
- Genetics
- Developmental Biology
Background:
- Endothelial cells are regulated by their microenvironment, which is lost in traditional cell isolation methods.
- Studying endothelial cells in their native environment is crucial for understanding their function.
Purpose of the Study:
- To develop a method for profiling gene expression in undisturbed endothelial cells in vivo.
- To identify novel endothelial genes and organ-specific expression patterns.
Main Methods:
- Developed the "AngioTag" zebrafish transgenic line.
- Utilized Translating Ribosome Affinity Purification (TRAP) coupled with RNA sequencing (TRAP-RNAseq).
- Generated a "UAS: RiboTag" line for broader cell type analysis.
Main Results:
- Successfully isolated and profiled actively translating mRNAs from endothelial cells in vivo.
- Demonstrated strong enrichment of endothelial-specific genes.
- Discovered novel endothelial genes and unique gene expression signatures across different adult organs.
Conclusions:
- The AngioTag and UAS: RiboTag systems provide powerful tools for studying cell-specific gene expression in vivo.
- These methods offer an unparalleled resource for understanding the molecular identity of cells in their native context.
