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Multiple cis-regulatory elements control prox1a expression in distinct lymphatic vascular beds.

Virginia Panara1,2, Hujun Yu3,4, Di Peng1

  • 1Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala 75185, Sweden.

Development (Cambridge, England)
|May 9, 2024
PubMed
Summary

Researchers uncovered how Prox1 gene expression is controlled in lymphatic endothelial cells (LECs) during development. They identified specific DNA regions, called enhancers, that regulate Prox1, impacting lymphatic vessel formation and function.

Keywords:
EnhancersEvolutionary conservationGene regulationLymphatic endothelial cellProx1Transcription factorZebrafish

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Area of Science:

  • Developmental Biology
  • Molecular Biology
  • Genetics

Background:

  • Prospero-related homeobox 1 (Prox1) is a key gene for lymphatic endothelial cell (LEC) development and lymphatic vasculature formation.
  • While Prox1 is essential, its upstream regulatory mechanisms in LECs are not fully understood.
  • Previous studies suggested SOX18 and COUP-TFII regulate Prox1 in mice, but detailed regulation in LECs requires further investigation.

Purpose of the Study:

  • To identify and characterize regulatory elements controlling Prox1 expression in zebrafish lymphatic endothelial cells.
  • To understand the role of these regulatory elements in lymphatic development and function.

Main Methods:

  • Utilized evolutionary conservation analysis and chromatin accessibility data to predict potential enhancers near the zebrafish prox1a gene.
  • Employed CRISPR/Cas9 gene editing to mutate a specific lymphatic valve enhancer.
  • Assessed the impact of enhancer deletion on lymphatic valve morphology and function.

Main Results:

  • Identified multiple enhancers near the zebrafish prox1a gene that are active in developing LECs.
  • Demonstrated that deletion of a lymphatic valve enhancer leads to impaired valve morphology and function.
  • Showed that distinct sets of enhancers control Prox1 expression: distal enhancers drive broad lymphatic expression, while proximal enhancers refine expression in specific lymphatic subsets.

Conclusions:

  • Prox1a expression in zebrafish is intricately controlled by a network of enhancers.
  • Ray-finned fish-specific distal enhancers are crucial for pan-lymphatic expression.
  • Vertebrate-conserved proximal enhancers play a role in refining Prox1a expression for specific lymphatic endothelial cell populations and functions.