Developing a novel zebrafish model to study mechanisms of neurodegeneration in pontocerebellar hypoplasia

Pontocerebellar hypoplasia (PCH) is a group of autosomal recessive neurodegenerative disorders with prenatal onset. The common characteristics are cerebellar hypoplasia with variable atrophy of the cerebellum and the ventral pons. The genetic basis of PCH is not fully understood, however a subset of patients have mutations in genes involved in tRNA splicing and maturation.

Rare autosomal recessive disorders such as PCH are commonly associated with consanguinity. Consanguinity often occurs in the UK as a consequence of sociocultural factors. In the Born in Bradford study we highlighted the increased prevalence of congenital disorders in babies of Pakistani origin:
http://www.thelancet.com/journals/lancet/article/PIIS0140-6736(13)61132-0/abstract
This was associated with high rates of consanguinity and first cousin marriage. By studying these families we have identified the genetic basis for several autosomal recessive disorders e.g. http://www.nature.com/ng/journal/v46/n5/full/ng.2948.html
http://www.nature.com/doifinder/10.1038/ng.2851
We have recently identified a novel PCH gene in a consanguineous family. Although the function of this gene is partially understood, we do not know how this specific mutation leads to PCH.

Zebrafish offer an excellent model system to confirm the disease gene in the context of a vertebrate whole organism, and to investigate the disease mechanisms. During this PhD you will develop a novel zebrafish model of PCH using gene-editing approaches. To characterise the model you will determine levels of survival of mutant embryos, and use molecular probes to investigate cerebellar hypoplasia. You will determine whether this phenotype can be rescued by injecting wild type or mutant mRNA into zebrafish embryos. To investigate whether the new gene also has a function in tRNA splicing and maturation you will determine the effect of genetic crosses with zebrafish carrying mutations in other PCH genes (TSEN54, CLP1) on survival of mutant zebrafish

To translate these findings to the clinic you will use high throughput in situ hybridization and tissue specific EGFP-reporter genes to develop a quantitative imaging-based screen for cerebellar hypoplasia in zebrafish which can then be used to screen for pharmacological modifiers of the PCH cerebellar phenotype in vivo.