Aging Markers Based On Oxidation Of Macromolecules

Aerobic metabolism and the corresponding generation of ROS is the most widely recognized cause of aging, although many questions remain regarding the detailed mechanism of action of ROS in organismal aging (Chien and Karsenty, 2005; Hadley et al., 2005). Several lines of evidence argue that macromolecular oxidation by ROS is responsible for functional declines or inappropriate roles of proteins, lipids, and DNA (Balaban et al., 2005). It seems likely that the accumulation of these oxidized dysfunctional macromolecules becomes a hallmark of progression of aging in zebrafish, as commonly recognized in other aging animals.

Protein oxidation has been detected by estimating its carbonyl content by Western blotting analysis (Dalle-Donne et al., 2003). In current studies, we have identified increasing accumulation of oxidized proteins with age in skin, muscle, and brain of zebrafish, when we examined the samples from 6-30-month-old fish. However, we have not detected difference in ovary where we observed a constant amount of oxidized proteins. We have also measured the accumulation of the oxidized lipid by-product, lipofuscin, as the "age pigment,'' one of the most prominent age-related cellular alterations reported in many organisms (Brunk and Terman, 2002; Terman and Brunk, 2004). Although in our initial studies, we did not detect lipofuscin accumulation in 24-month old fish, our currently updated results indicate that gradually increased accumulation of lipo-fuscin occurs at 30 months and even more significantly at 42-months. In addition to muscle tissues, lipofuscin accumulation was observed in brain, liver, kidney, but not intestine, with advancing age. For each experiment, we examined 10-15 fish samples, and quantified the results by image intensities of Western blotting for oxidized proteins and by capturing images to count fluorescent spots for lipofuscin. Statistical analysis of these oxidative-stress-associated markers is currently in progress, but the relevance of the markers is already evident.

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Blood Pressure Health

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