Developing Tools to Investigate VEGFA Signaling in Brain Vascular Development

The central nervous system (CNS) relies on a tightly regulated vascular network to support its development, maintain its homeostasis, and respond to disease. Blood vessels, lined with endothelial cells, deliver oxygen and nutrients to the brain. Vascular endothelial growth factor A (VEGFA) is the main growth factor necessary for vascular development, and its function is conserved in zebrafish. VEGFA signalling regulates endothelial cell differentiation, proliferation, and migration. VEGFA blockers are used in clinical settings to treat cancer and hypervascularization conditions.

This project aimed to establish tools to study VEGFA signalling during zebrafish brain vascular development. We used a new genome editing technique, CRISPR/Cas9 based mutagenesis tool kit, to simultaneously generate a reporter of vegfa and disrupt its expression. We observed the expression pattern of the fluorescent reporter in the trunk, somites, and occasionally the head, recapitulating its known endogenous expression. Whole mount vegfa immunostaining indicate expression primarily in the head region, while immunostaining of adult brain sections didn’t show a specific signal. Finally, we characterized a glut1b antibody specific to brain blood vessels, enabling future investigation of vegfa inhibition. Together, this work establishes foundational tools and validate zebrafish as a practical model for studying VEGFA signaling in brain vascular development.