Dissecting the mechanisms controlling basement membrane formation and stability with Drosophila genetics (360G-Wellcome-301803_Z_23_Z)

The basement membrane (BM) is a multi-component polymer network underlying all epithelia and surrounding many tissues. Despite its ubiquity, we have little understanding how its components are polymerised, interact, and are maintained throughout life. While the BM is often assumed to be a static structure, we hypothesise that its formation is a dynamic, developmentally controlled process, and that investigating progressive formation from an immature embryonic network through to a mature, homeostatic state will reveal essential regulatory steps in BM assembly and maintenance. We will utilise a mass spectrometry-based pipeline to comprehensively identify BM-associated components, posttranslational modifications, and alterations in BM stability throughout Drosophila development to understand how the network progressively matures to acquire a final stable structure. We will simultaneously establish imaging approaches to examine the changes in rates of BM turnover during the maturation process. Finally, we will exploit our mass spectrometry and imaging techniques, along with the genetic tractability of flies, to functionally dissect BM molecular alterations and the developmental signals controlling BM maturation. Our unique ability to biochemically characterise BM formation and live image component dynamics within a living animal will elucidate how this complex polymer network functions during normal animal physiology and disease states.