Dissecting the role of Hedgehog signalling during development, regeneration and ageing in vivo using novel phosphorescent small molecules

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The goal of this PhD project is to study the changing roles of Hedgehog signalling during amphioxus' entire lifespan, from embryonic development to adult regeneration, using novel phosphorescent compounds. The primary aims include: 1. to characterise phenotypes resulting from treatment of chemical compounds affecting Hedgehog signalling via immunohistochemistry and whole mount in situ hybridisation 2. to generate novel iridium tagged compounds that can be used to image the Hh signalling pathway. The current way of imaging (immunofluorescence) has the following limitations: it is hard to quantify, most cells and organisms naturally autofluoresce which makes specific fluorescent labels hard to visualise, and they are issues with using them to image live organisms. The novel iridium tagged compound we plan to create needs to be the following: water soluble and biocompatible (to allow in vivo imaging). We are planning on using a phosphorescent cationic iridium(III) complex and we hope to develop this further to create a colour palette.3. to generate novel iridium tagged compounds with photoconvertible properties in order to modulate spatial and temporal activity of the biomolecules. Amphioxus is the model organism used for this project due to the following: it shares many similarities with vertebrates but has a simpler genome and anatomy (making the Hedgehog signalling pathway easier to study), it is semi-translucent (aids live imaging), it regenerates well and being an invertebrate chordate makes it a great ethical alternative to adult regeneration research in vertebrates. This project has clinical applications due to the Hedgehog signalling pathway having links to neural defects, skeletal defects, cyclopia, cancer and ciliopathies. In addition, the study of regeneration is closely linked to wound healing so can help our understanding of hedgehog signalling in both regeneration and wound healing.