Cytoneme-mediated morphogen transport during wound healing and regeneration (ref: 4270)

The determination of the cellular fate is one of the most fundamental and classical problems in regeneration. In many tissues, cell fate is determined by morphogen signalling proteins.
Two families of morphogens, namely Shh and Wnt - are essential for determining many fundamental aspects such as cell proliferation, differentiation and migration. We and others have shown that the distribution of these morphogens can be facilitated by membrane protrusions - also known as cytonemes. Cytonemes are thin, actin-rich, and transport essential components of the Shh and Wnt signalling pathway between cells (Zhang and Scholpp, 2019; Curr Opin Gen & Dev). Cytonemes are highly dynamic and can form and retract within minutes (Stanganello et al., 2015; Nat Comms).
Their emergence is precisely controlled by the cytoneme-producing cell and the extracellular space through which they traverse (Brunt, et al., 2021, Nat Comms). Although there is strong evidence that Wnt3a and Shh cooperate during wound healing, to date, it is unknown how their crosstalk is regulated. Furthermore, the role of cytoneme-mediated transport during wound healing is unclear.
Under the supervision of leading cell and developmental biologists, the student will study Wnt and Shh transport in the regenerating fin in zebrafish. The student will generate transgenic zebrafish lines in which signalling components are fluorescently tagged to investigate cytoneme-based morphogen release by quantitative imaging. We will also use real-time PCR, single-molecule fluorescence in-situ hybridisation, and advanced high-resolution microscopy in the wound to describe signal activation. Finally, the student will interfere with the emergence of cytonemes by using chemical inhibitors and CRISPR/Cas9-based mutations to study their impact on zebrafish wound healing.