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Title: Matrix regulatory genes in the human lens
Author: Hodgkinson, Lisa Marie
ISNI:       0000 0001 3579 7716
Awarding Body: University of East Anglia
Current Institution: University of East Anglia
Date of Award: 2008
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Purpose: The proteinases that cells employ to regulate the matrix include members of the zinc metalloproteinases; expression is likely to influence both the normal and injured human lens, which in tum can affect a number of biological functions e.g. migration. The current study therefore aimed to determine expression profiles in distinct regions ofthe human lens and following mechanical injury. Methods: For specific gene expression analyses the native human lens was dissected into three regions, using sham cataract surgical methods. Primary lens epithelial cell culture provided cells for analysis as a wounded lens system. Native lens capsular bags (CB) were cultured to assess short term wounding responses and ex vivo CBs (obtained with implanted intraocular lenses from cataract surgery sometime prior to death), were analysed to study the long term wounding responses. Total RNA extraction and reverse transcription technologies were developed for real time polymerase chain reaction amplification gene expression analysis. Protein expression was investigated using Western blotting techniques. Results: In the native lens, the overall MMP gene expression was low, while TIMP expression was high. Of the MMP family members, MTI-MMP was the most highly expressed in the epithelium, while MT2-MMP was the most abundant gene in the fibre cells. Moreover, the presence of unprocessed and activated MTI and MT2-MMP were confirmed by Western blotting. Selected adamalysin genes (ADAM-9, -10, -15 and -17) were highly expressed by all native lens regions. In the wounded lens and ex vivo samples transdifferentiation marker expression; alpha smooth muscle actin and fibronectin, was consistent with the different stages following cataract surgery. Significant elevation of specific gene expression was observed follo~ving injury, such as MMP-2, MMP-9, ADAM-9, ADAM-IS and ADAMTS-3. Conclusions: The present study demonstrates that regional distribution of MMPs, TIMPs, ADAMs and ADAMTSs occurs within the human lens. In the native lens expression of matrix degrading proteinases is generally low, while inhibitor expression is high. Following surgical injury, a significant increase in levels of specific genes, such as MMP-2 is observed; these particular genes are therefore likely to play key roles in the wound healing process after cataract surgery and are thus logical subjects for further investigation.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available