Age-estimations of rats based on the average lateral diffusion constant of hepatocyte membrane proteins as revealed by fluorescence recovery after photobleaching.

A method has been developed recently for measuring the average lateral diffusion constant of the proteins (D) in the cell membrane of hepatocytes in liver smears by fluorescence recovery after photobleaching (FRAP). A peroxide-induced autofluorescence (PIAF) of the membrane proteins was used as a fluorescent label. It has been established that D displays a significant negative linear correlation with age. The present paper describes age-estimations carried out on 12 male Fischer 344 rats (7-29 months of age) in so-called "blind experiments": the operator knew only the sex of the rat, determined D from a small piece of the freshly removed liver, and estimated the age of the rat from the age-dependent regression line for D established previously on 16 other Fischer 344 male rats of various ages. There was a strong correlation of the estimated age with the actual one (r = 0.92), the slope of the regression line was 0.98 and its intercept differed from 0 by only 0.5 months. These results indicate that D may play a decisive role in the determination of membrane functions as predicted by the membrane hypothesis of aging.