Hu, W; (2014) Characterisation of corneal development in 'Xenopus laevis' and functional analysis of 'XTgfbi'. Doctoral thesis , UCL (University College London).

Abstract

Corneal dystrophies are a group of inherited eye diseases that affect corneal transparency in humans. I am interested in a subgroup of human corneal dystrophies caused by mutations in the gene Transforming growth factor β-induced (TGFBI). In order to explore the possibility of using Xenopus laevis as a model system to study TGFBI-associated corneal dystrophies, I carried out investigations into Xenopus corneal development, the role of Xenopus TGFBI (XTgfbi) during embryogenesis and the expression pattern of XTgfbi in cornea. The thesis first describes the full process of corneal development in Xenopus laevis. Both light and electron microscopy were used to illustrate the overall structure of the cornea, as well as the subcellular details, throughout the course of development. Interesting structures such as a stroma-attracting centre, multilayered double embryonic cornea and intra-stromal epithelium were described. Overall, Xenopus corneal development is highly complex, and its similarities and differences with that in other model organisms will be discussed in depth. In addition, I performed proliferation assays and found cells with proliferative capacity in all three cellular layers in the mature cornea. The second part of the thesis focuses on the functional analysis of extracellular matrix protein XTgfbi during embryogenesis. After confirming previous data of gene expression and knock-down phenotypes, I showed that XTgfbi specifically regulates the canonical Wnt signalling pathway using semi-quantitative reverse transcription – polymerase chain reaction (RT-PCR), confirmed by luciferase reporter assays. A final set of experiments in this section suggests integrin-linked kinase might mediate XTgfbi’s function in Wnt signalling regulation. The third part of the thesis describes the expression pattern of XTgfbi during Xenopus corneal development. It was found that XTgfbi was present in different types of corneal cells during the course of development. The dynamic nature of its expression pattern implicates its complex yet important roles in this process.

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