Lineage analysis of the ENS precursors in the zebrafish. Taking advantage of the optical clarity of the zebrafish embryo and the accessibility and manipulability of the premigratory neural crest we are genetically labeling subpopulations of the vagal neural crest-derived enteric neural crest cells (ENCCs) to determine if there are distinct subpopulations of ENCCs that give rise to different subtypes of ENS neurons.
The in vivo function of zebrafish orthologues of known HSCR genes. Taking advantage of the zebrafish model system we are investigating how known HSCR linked genes, such as Sip1, cause their ENS phenotypes. We have previously shown that the neurotrophic factor GDNF and its receptor complex are absolutely required for normal ENS development in zebrafish as in mouse and human. Using similar techniques we are investigating the function of other known HSCR genes in zebrafish ENS development. These studies are investigating the many unanswered questions as how these known HSCR genes actually cause their ENS phenotypes when mutated.
Analyzing in vivo function of new potential HSCR loci using zebrafish. Taking advantage of the zebrafish model system we are investigating whether zebrafish orthologues of genes identified in HSCR patient exome sequencing are involved in ENS development in the zebrafish. These studies hope to identify novel HSCR loci.
The relationship between gut endoderm development and ENS development. Our studies of the zebrafish ENS mutants suggest that factors derived from the gut endoderm play a critical role in the normal migration and proliferation of ENS precursors. We are taking a microarray approach to determine which gut endoderm factors are downregulated in these mutants.
Development of FACS sorting ENS precursor cells for RNA seq and Chip Seq studies.
Development of a small molecule screen utilizing double transgenic Zebrafish to identify novel compounds that promote or inhibit ENS precursor proliferation and/or differentiation.
