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Apoptosis gene expression profiling in lens development

Geatrell, Jenny Claire 2007. Apoptosis gene expression profiling in lens development. PhD Thesis, Cardiff University.

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Abstract

During lens development, epithelial cells located at the equatorial region of the lens undergo terminal differentiation to form fibre cells resulting in cell elongation and degradation of all intracellular organelles. Failure to complete this process successfully can result in cataract. This process is thought to be an attenuated form of classical apoptosis. This study was completed to give a comprehensive analysis of apoptosis genes in the developing mouse lens. Macroarrays containing 243 immobilised cDNAs with a known role in apoptosis were utilised to examine the gene expression at different developmental stages. The stages examined were postnatal day 7 and postnatal day 14. Over 100 apoptosis genes were shown to be expressed above background with 20 genes demonstrating significantly differential expression (2-fold or greater change in expression, p-value < 0.05), with highest expression at PI4. PCR confirmed expression of all the genes identified from the array results, and differential expression was confirmed for 52% of the genes. Protein expression of two selected genes, axl and mcl-1, was demonstrated using western blotting. Lens morphology was examined in transgenic mice generated to contain an extended CAG repeat in the huntingtin gene (one of the genes identified from the arrays). Morphology was compared between homozygote, heterozygote and wild-type mice. The presence of the mutated gene did not affect denucleation during lens differentiation and no statistically significant difference was seen in the dimensions of the organelle free zone (OFZ) of the wild-type and homozygote mice. A cross-species comparison was completed. Gene expression of the genes shown to be highly expressed and differentially expressed from the array results was examined in embryonic chick lenses. From the results of this part of the study expression was seen for 83% of genes. These results add to the argument that the process of differentiation is similar in both mouse and chick lenses.

Item Type: Thesis (PhD)
Status: Unpublished
Schools: Optometry and Vision Sciences
Subjects: R Medicine > RE Ophthalmology
ISBN: 9781303209857
Date of First Compliant Deposit: 30 March 2016
Last Modified: 25 Oct 2017 14:28
URI: http://orca-mwe.cf.ac.uk/id/eprint/54644

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