mRNAs are found not only in neuronal cell bodies but also in axons and dendrites, where many re locally translated. These local mechanisms are important for proper neuronal development and adult neurological functions. Alternative splicing of mRNAs produces various protein isoforms and regulates neuronal maturation and connectivity. The Houart lab has identified several RNA splicing proteins that are localised to growing and maturing axons, where they regulate the local transcriptome and control axonal shape and connectivity. Interestingly, nucleo/cytoplasmic distribution of RNA splicing proteins have been found disrupted in neurodegenerative diseases such as ALS.
The mRNAs coding for the core splicing protein SRSF1 are localised to, and translated in, axons of embryonic zebrafish spinal cord neurons. The aim of this project is to investigate interactions between axonal RNA splicing proteins and specific mRNAs during neuronal development, focusing on SRSF1.
The candidate will work to understand these interactions – such as when they occur during neuronal development and where they occur within the cell – and how they impact neuronal development.
Alongside standard cell and molecular biology techniques, skills that will be learnt include: immunostaining, in situ hybridisation, generating zebrafish reporter and mutant lines, microinjection, electroporation, proximity labelling, RNAseq, neuronal cell culture, and confocal, lightsheet and expansion microscopy including in vivo timelapse imaging.
Objectives:
Generate (Year 1) and characterise (Year 2) zebrafish with axon-specific mutation for RNA splicing protein SRSF1
Investigate timing and dynamics of SRSF1 localisation in axons (Year 1)
Determine molecules that SRSF1 interacts with in the axon (mRNAs and other proteins – Year 2 and 3)