Dynamic BMP signaling mediates notochord segmentation in zebrafish

Abstract

The vertebrate spine is a metameric structure composed of alternating vertebral bodies (centra) and intervertebral discs.¹ Recent studies in zebrafish have shown that the epithelial sheath surrounding the notochord differentiates into alternating cartilage-like (col2a1/col9a2+) and mineralizing (entpd5a+) segments which serve as a blueprint for centra formation.^2,3,4,5 This process also defines the trajectories of migrating sclerotomal cells that form the mature vertebral bodies.⁴ Previous work demonstrated that notochord segmentation is typically sequential and involves the segmented activation of Notch signaling.² However, it is unclear how Notch is activated in an alternating and sequential fashion. Furthermore, the molecular components that define segment size, regulate segment growth, and produce sharp segment boundaries have not been identified. In this study, we uncover that a BMP signaling wave acts upstream of Notch during zebrafish notochord segmentation. Using genetically encoded reporters of BMP activity and signaling pathway components, we show that BMP signaling is dynamic as axial patterning progresses, leading to the sequential formation of mineralizing domains in the notochord sheath. Genetic manipulations reveal that type I BMP receptor activation is sufficient to ectopically trigger Notch signaling. Moreover, loss of Bmpr1ba and Bmpr1aa or Bmp3 function disrupts ordered segment formation and growth, which is recapitulated by notochord-specific overexpression of the BMP antagonist, Noggin3. Our data suggest that BMP signaling in the notochord sheath precedes Notch activation and instructs segment growth, facilitating proper spine morphogenesis.

Keywords: BMP signaling; Bmp3; Nog3; Notch; mineralization; notochord segmentation; zebrafish.