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A.R. Prescott, A. Sandilands, J. James, J. Kuszak, M. Pekny, A. Chepelinsky, A. Wegener, R. Zoltoski, A. Määttä, R. Quinlan; Optical and Anatomical Analysis of Dundee Mice . Invest. Ophthalmol. Vis. Sci. 2004;45(13):4600.
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Purpose: To identify the function of CP49 in lens fibre cell organisation and lens optical properties Methods: We have generated a CP49 targeted knockout and determined that light scatter and lens optical properties had deteriorated in the CP49 knockout lenses compared to litter mate controls. Detailed morphological examinations revealed that these changes were accompanied by dramatic alterations in plasma membrane organisation of the fibre cells. A CP49–vimentin and filensin knockouts were also generated. The CP49 knockout mouse is an important model to study the functional link between lens transparency and function, the cytoskeleton and plasma membrane organisation. Lenses were analysed by slit lamp and by helium neon laser scan analysis to measure the average back focal length (BFL; spherical aberration) and back focal length variability (BFLV; sharpness of focus) of knockout and wild–type littermate lenses. Lenses were sectioned and then processed for confocal immunofluorescence microscopy and immunoelectron microscopy. Results: Targeted removal of CP49 resulted in the degradation of lens optical quality. A strain dependence (129X1 versus C57Bl6) was observed, but the effect upon the cytoskeleton appeared consistent between the two strains. The effect of CP49 deletion upon a selection of membrane markers (eg AQP0, periplakin, plectin) was investigated by immunofluorecence and immunoelectron microscopy. The organisation of these markers changed as a result of the loss of CP49 and thebeaded filament cytoskeleton. Conclusions: The targeted deletion of CP49 dramatically alters lens fibre cell organisation, both in terms of the cytoplasmic cytoskeleton and the plasma membrane cytoskeleton complex. These changes correlate with altered optical properties.
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