May 2004
Volume 45, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2004
Alpha–Crystallin prevents cellular disintegration during the apoptosis–like program of lens fiber cell maturation.
Author Affiliations & Notes
  • V. Morozov
    Laboratory of Molecular and Developmental Biology, NEI, NIH, DHHS, Bethesda, MD
  • E.F. Wawrousek
    Laboratory of Molecular and Developmental Biology, NEI, NIH, DHHS, Bethesda, MD
  • Footnotes
    Commercial Relationships  V. Morozov, None; E.F. Wawrousek, None.
  • Footnotes
    Support  none
Investigative Ophthalmology & Visual Science May 2004, Vol.45, 1150. doi:
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      V. Morozov, E.F. Wawrousek; Alpha–Crystallin prevents cellular disintegration during the apoptosis–like program of lens fiber cell maturation. . Invest. Ophthalmol. Vis. Sci. 2004;45(13):1150.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Abstract: : Purpose: To test the hypothesis that the absence of alpha–crystallin in mouse lenses permits the apoptosis–like process of lens fiber cell maturation to progress beyond its normal stopping point, and leads to the cell disintegration observed in the lens cortex of alpha–crystallin null mice. Since it had been previously reported that alphaB–crystallin can inhibit the autocatalytic maturation of the execution caspase 3, we postulated that in the normal lens, alpha–crystallin prevents the final steps of the apoptosis process, maintaining cellular integrity after subcellular organelle destruction. Methods: alphaA/alphaB–crystallin double knockout (DKO) mice were previously generated in this lab. Lens cell extracts from wild type (WT) and DKO mice were assayed with commercially available kits, in the presence of several specific protease inhibitors, to determine the activities of individual members of the caspase superfamily. Immunofluorescence studies were conducted on frozen lens sections and viewed with a standard epiflourescence. Results: Among those studied, caspase 3 and caspase 6, both belong to the execution caspase subfamily, exhibited elevated activity in DKO lenses, as compared to WT. Lens extracts from various ages of DKO mice exhibited two to four fold higher caspase 3 activity than WT. Similarly, the caspase 6 activity in DKO mouse lens extract was two fold higher than in WT. Immunofluorescence microscopy revealed an increase in the active form of caspase 3 in specific regions of the DKO lens, consistent with the site of cell disintegration. Conclusions: In the absence of alpha–crystallin, lens fiber cells fail to halt their apoptosis–like maturation program, and progress to the stage of cell disintegration. Increased activities of apoptotic execution caspases in these DKO lenses contribute to the cell disintegration, consistent with alpha–crystallin inhibition of caspase activity in the normal lens.

Keywords: apoptosis/cell death • crystallins • transgenics/knock–outs 
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