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V.R. Harrington, O.P. Srivastava; Localization of -Crystallins in Aging Normal and Cataractous Human Lenses . Invest. Ophthalmol. Vis. Sci. 2003;44(13):2364.
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Purpose: The objective of this study was to determine the localization of truncated α-crystallin in aging human lenses and carry out comparative localization of the crystallin fragments in normal and age-matched nuclear cataractous human lenses. Methods: Fresh 87-year-old normal and 79-year-old cataractous lenses were fixed, cryoprotected, and sectioned at 16 µm using the Leica 2800 E Friocut cryostat. The immunohistochemical analysis was done using two primary antibodies i.e., anti-αA-N-(residue nos. 1-9) antibody and anti-αA-C-(residue no. 165-173) antibody. The secondary antibody was anti-rabbit IgG, made in goat and the fluorescent dye used for staining was Alexa 594 (red). Results: The results showed no difference in distribution of αA-crystallin in the central region on immunoreactivity with either anti-N-terminal or anti-C-terminal antibodies. The 87-year-old normal lens show a strong intense staining in the peripheral and bow regions with the anti-αA-N-antibody compared to the control. No such immunoreactivity with anti-αA-C antibody was observed. Similarly, the 79-year-old cataractous lens showed staining in the peripheral region only with the anti-αA-N-antibody. Conclusions: The lack of immunoreactivity with the anti-C-terminal antibody suggests loss of the C-terminal region in αA-crystallin in the peripheral region of both normal and cataractous lenses..
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