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We have generated two mouse models, in one by inserting the human lens αAN101D transgene in CRYαA N101D  mice, and in the other by inserting human wild-type αA-transgene in CRYαA WT  mice. The CRYαA N101D  mice developed cortical cataract at about 7-months of age relative to CRYαA WT  mice. The objective of the study was to determine the following relative changes in the lenses of CRYαA N101D - vs. CRYαA WT  mice: age-related changes with specific emphasis on protein insolubilization, relative membrane-association of αA N101D  vs. WTαA proteins, and changes in intracellular ionic imbalance and membrane organization.    Methods  Lenses of varying ages from CRYαA WT  and CRYαA N101D  mice were compared for an age-related protein insolubilization. The relative lens membrane-association of the αAN101D- and WTαA proteins in the two types of mice was determined by immunohistochemical-, immunogold-labeling-, and western blot analyses. The relative levels of membrane-binding of recombinant αA N101D - and WTαA proteins was determined by an in vitro assay, and the levels of intracellular Ca 2+  uptake and Na, K-ATPase mRNA were determined in the cultured epithelial cells from lenses of the two types of mice.    Results  Compared to the lenses of CRYαA WT , the lenses of CRYαA N101D  mice exhibited: (A) An increase in age-related protein insolubilization beginning at about 4-months of age. (B) A greater lens membrane-association of αAN101D- relative to WTαA protein during immunogold-labeling- and western blot analyses, including relatively a greater membrane swelling in the CRYαA N101D  lenses. (C) During in vitro assay, the greater levels of binding αAN101D- relative to WTαA protein to membranes was observed. (D) The 75% lower level of Na, K-ATPase mRNA but 1.5X greater Ca 2+  uptake were observed in cultured lens epithelial cells of CRYαA N101D-  than those of CRYαA WT  mice.    Conclusions  The results show that an increased lens membrane association of αA N101D - − relative WTαA protein in CRYαA N101D  mice than CRYαA WT  mice occurs, which causes intracellular ionic imbalance, and in turn, membrane swelling that potentially leads to cortical opacity.

bmc ophthalmology

Increased Association of Deamidated αA-N101D with Lens membrane of transgenic αAN101D vs. wild type αA mice: potential effects on intracellular ionic imbalance and membrane disorganization