Abstract
We developed a monoclonal antibody against HPLC purified rat lens gamma L crystallins. This antibody was specific to both the polypeptides (19,000 and 21,000 daltons) which constituted the HPLC gamma L peak. Least reactivity was shown against gamma H (24,000 daltons). This antibody was used as a probe to detect the presence of and quantitate gamma L crystallins in lens soluble, insoluble and urea-insoluble fractions during diabetes. Utilizing a direct binding immunoassay (ELISA) we calculated the absolute quantities of gamma L crystallins present in these fractions. Our results show, in normal animals there was a minimal change in total quantities of gamma L crystallins in soluble fraction from 1 month to 5 months of age, but a slow accumulation of these crystallins in insoluble and urea-insoluble fractions was seen during the same period. Diabetes resulted in a depletion of gamma L crystallins from the soluble fraction, both in terms of relative proportion and absolute quantities. In insoluble and urea-insoluble fractions the relative proportions of these crystallins were increased dramatically up until 60 days followed by a decrease during 90-120 days of diabetes, whereas, the absolute quantities remained more or less steady after reaching the maximum on 60 days. Although the relative proportions of gamma L crystallins in the insoluble fraction seem to be less when compared to urea-insoluble fraction, the total quantity of these crystallins was much higher due to abundance of this fraction.(ABSTRACT TRUNCATED AT 250 WORDS)
Published Version
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