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M.D. O'Connor, J.W. McAvoy; Reconstruction of the Mammalian Lens . Invest. Ophthalmol. Vis. Sci. 2004;45(13):2628.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: Previous lens explant studies within our laboratory resulted in the production of transparent tissues whose cellular organization resembled that of the anterior portion of the lens. The present study aimed to generate a lens–like tissue capable of focusing light in vitro, by mimicking the lens vesicle stage of lens development. Methods: The lens vesicle was approximated in vitro by setting up paired lens epithelial cell explants (ie. ‘explant–pairs’). Explant–pairs were cultured in vitreous for up to 45 days, after which the tissues were photographed and analyzed by light microscopy, electron microscopy, confocal microscopy and immunofluorescence. A proteomic analysis of lens membrane proteins was also undertaken, using 2D electrophoresis and mass spectrometry. Results: After approximately 30 days, the cultured explant–pairs were transparent, focused light and magnified images. Additionally, these tissues contained molecular and ultrastructural markers of both epithelial and fibre cells. When cultured for 45 days, the tissues developed a diffuse opacity biomicroscopically and ultrastructurally similar to that reported for human age–related, nuclear cataract. Preliminary analysis of the lens membrane proteins indicated the possible identification of phosphorylated FGF receptor 3. Conclusions: We have developed a transparent tissue capable of focusing light and magnifying images, which contains both epithelial– and fibre–like cells. Furthermore, these tissues develop an opacity similar to the human age–related, nuclear cataract. Due to the length of time required for cataract to develop in vivo as well as difficulties in obtaining specimens over this time, our novel system may become a valuable tool for investigating normal lens development as well as the mechanisms involved in cataract formation. To identify the vitreous factors required for lens development, we collected lens membrane proteins in the hope of obtaining activated growth factor receptors. Preliminary results show the possible identification of phosphorylated FGF receptor 3, indicating the potential for identification of other growth factor receptors using this approach.
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