The fate of gamma L crystallins in rat lens during diabetic cataractogenesis as determined by a monoclonal antibody.
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)
Swamy
M S
MS
Department of Cell and Molecular Biology, Medical College of Georgia, Augusta 30912-2100.
Shyamala
M
M
Abraham
J
J
Garver
F A
FA
Abraham
E C
EC
D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D015153:Blotting, Western; D002386:Cataract / Q000209:etiology / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D003459:Crystallins / Q000302:isolation & purification / Q000378:metabolism*; D003921:Diabetes Mellitus, Experimental / Q000150:complications / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D007908:Lens, Crystalline / Q000378:metabolism*; D008297:Male; D051381:Rats; D011919:Rats, Inbred Strains; D012995:Solubility
D000911:Antibodies, Monoclonal; D003459:Crystallins