Norton, William Howard James; (2002) Purinergic signalling during zebrafish development and the characterisation of novel fish mutants. Doctoral thesis (Ph.D), UCL (University College London).

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Abstract

This thesis looks at two different aspects of zebrafish development and as such is divided into two main sections. The first section looks at the expression of novel purinoceptors during zebrafish development. Purinoceptors are receptors that are activated by either adenosine or ATP and its analogues. They have been shown to have important roles during the development of many species. The thesis looks at the expression of three new zebrafish homologues, two of which are ionotropic receptors (p2x3 and p2x4) and one of which is a G protein coupled receptor (p2y11), both in wild-type embryos and embryos of selected mutants. The results suggest roles for these receptors during development and show that the fish is a good model to study early developmental expression patterns. Experiments to elucidate the function of one of the genes were performed. The second part of the thesis reports the finding and characterisation of novel zebrafish mutants. A large scale ENU screening strategy was used to identify a large number of fish with axonal pathfinding defects at 36 hours of development, using an anti-acetylated tubulin antibody. The embryos were screened for defects in the axon scaffold formation. The final results chapter describes the characterisation of monorail, a zebrafish midline mutant that shows floorplate defects. A combination of midline marker gene expression pattern analysis and mapping of the mutation to known single sequence length polymorphisms (SSLPs) allows the mutated gene to be identified.

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