H₂S promotes developmental brain angiogenesis via the NOS/NO pathway in zebrafish

Hydrogen sulphide (H₂S) is considered as the third member of the gasotransmitter family, along with nitric oxide (NO) and carbon monoxide. H₂S has been reported to induce angiogenesis by promoting the growth, migration and tube-like structure formation of endothelial cells. Those studies were conducted in conditions of cell culture, mouse Matrigel plug assay model, rat wound healing model or rat hindlimb ischaemia model. Recent in vivo studies showed the physiological importance of H₂S in muscle angiogenesis. However, the importance of endogenous H₂S for brain angiogenesis during development remains unknown. We therefore aimed at determining the role of H₂S in brain vascular development.

Both knockdown and knockout of H₂S-producing enzymes, cystathionine β-synthase (cbs) and cystathionine γ-lyase (cth), using morpholino oligonucleotides and clustered regularly interspaced short palindromic repeats/Cas9-mediated mutation, impaired brain vascular development of larval zebrafish. Incubation with the slow-releasing H₂S donor GYY4137 alleviated the defects of brain vascular development in cbs and cth morphants. Quantitative analysis of the midbrain vascular network showed that H₂S enhances angiogenesis without affecting the topological structure of the brain vasculature. Mechanically, nitric oxide synthase 2a (nos2a) expression and NO production were decreased in both cbs and cth morphants. Overexpression of nos2a by coinjection of cbs or cth MO with full-length zebrafish nos2a mRNA alleviated the brain vascular developmental defects in cbs and cth morphants.

We conclude that H₂S promotes brain developmental angiogenesis via the NOS/NO pathway in zebrafish.