Regulation of Hindbrain Vascular Development by rps20 in Zebrafish

During aging, the brain vasculature undergoes significant deterioration characterized by increased arterial tortuosity, compromised blood-brain barrier integrity, and reduced cerebral blood flow, all of which contribute to various neurological disorders. Thus, understanding the mechanisms underlying aging-related cerebrovascular defects is critical for developing strategies to alleviate aging-associated neurological diseases. In this study, we investigated the role of aging-related genes in brain vascular development using zebrafish as an in vivo model. By thoroughly analyzing scRNA-seq datasets of mid- and old-aged brain vascular endothelial cells (human/mouse), we found <i>ribosomal protein S20 (rps20)</i> significantly down-regulated during aging. qPCR analysis and whole-mount in situ hybridization validated a high expression of <i>rps20</i> during early zebrafish development, which progressively decreased in adult and aged zebrafish brains. Functional studies using the CRISPR/Cas9-mediated knockout of <i>rps20</i> revealed an impaired growth of central arteries in the hindbrain and a marked increased intracranial hemorrhage incidence. Mechanistically, qPCR analysis demonstrated a significant downregulation of <i>vegfa, cxcl12b, and cxcr4a,</i> key signaling molecules required for hindbrain vascular development, in <i>rps20</i>-deficient embryos. In conclusion, our findings demonstrate that <i>rps20</i> is essential for proper brain vascular development and the maintenance of vascular homeostasis in zebrafish, revealing a novel mechanism by which aging-related genes regulate brain vascular development. This study provides new insights that may aid in understanding and treating aging-associated vascular malformations and neurological pathologies.