Zebrafish glial-vascular interactions progressively expand over the course of brain development.

Glial-vascular interactions are critical for the formation and maintenance of brain blood vessels and the blood-brain barrier (BBB) in mammals, but their role in the zebrafish BBB remains unclear. Using three glial gene promoters-, , and (a truncated )-we explored glial-vascular development in zebrafish. Sparse labeling showed fewer glial-vascular interactions at early stages, with glial coverage and contact area increasing with age. Stable transgenic lines for and revealed similar developmental increases, starting at ∼30% coverage at 3 days post-fertilization (dpf) and peaking at ∼60% by 10 dpf, and consistently higher glial coverage in the forebrain and midbrain than in the hindbrain. Electron microscopy analyses showed similar progressive increases in glial-vascular interactions, with maximal coverage of ∼70% in adults-significantly lower than the ∼100% seen in mammals. These findings define the temporal and regional maturation of glial-vascular interactions in zebrafish and highlight differences from mammalian systems.