Identifying regulators of tissue regeneration by in vivo imaging in the zebrafish

This proposal aims to use a zebrafish model of muscle regeneration in order to identify the signals between muscle stem cells (muSCs) and inflammatory cells. The purpose is to determine which signals regulate their interactions during regeneration and so control muscle repair. Specific Aims to be tested include:
1. determine how key pro- and anti-regenerative signals regulate muSCs and mø responses to tissue injury
2. identify candidate signalling pathways controlling muSCs and mø interactions during muscle regeneration
3. functionally test the role of candidate cell signalling pathways in controlling muSC responses to injury

To address these aims we will use transgenic zebrafish for gene over-expression and visualisation of cells during regeneration. Imaging of stem cell and macrophage responses to injury will be performed by confocal and multiphoton microscopy. To identify complementary signals and receptors expressed between muSCs and inflammatory cells, gene expression profiling will be performed on cells isolated by flow cytometry using RNA sequencing. Candidate secreted proteins and their respective receptors that could act to signal between muSCs and inflammatory cells will be identified using computational analyses of sequencing data. These candidate molecules will then be tested in vivo by over-expression under the control of tissue specific promoters and their affects measured by live cell imaging.

Please note: Applicants must include the project ID number (2017/DI/03) in the ’Research proposal’ and ’Funding (point 5)’ sections of their application.