Organization of the apical extracellular matrix during tubular organ formation

Abstract The apical extracellular matrix (aECM) plays a critical role in epithelial tube morphogenesis during organ formation, but its composition and organization remain poorly understood. Using the Drosophila embryonic salivary gland (SG) as a model, we identify Papss, an enzyme that synthesizes the universal sulfate donor PAPS, as a critical regulator of tube lumen expansion. Papss mutants show a disorganized apical membrane, condensed aECM, and disruptions in Golgi structures and intracellular trafficking. SG-specific expression of wild-type Papss, but not the catalytically inactive form, rescues the defects in Papss mutants, suggesting that defects in sulfation are the underlying cause of the phenotypes. Additionally, we identify two zona pellucida (ZP) domain proteins, Piopio (Pio) and Dumpy (Dpy), as key components of the SG aECM. In the absence of Papss, Pio is gradually lost in the aECM, while the Dpy-positive aECM structure is condensed and dissociates from the apical membrane, leading to a thin lumen. Mutations in dpy or pio, or in Notopleural, which encodes a matriptase that cleaves Pio to form the luminal Pio pool, result in a SG lumen with alternating bulges and constrictions, with the loss of pio leading to the loss of Dpy in the lumen. Our findings underscore the essential role of sulfation in organizing the aECM during tubular organ formation and highlight the mechanical support provided by ZP domain proteins in maintaining luminal diameter.