Calorie restriction (CR) in the rotifer Brachionus plicatilis extends its lifespan, as it enhances the expression of antioxidant enzymes such as manganese superoxide dismutase (Mn SOD) and catalase. Here we show that CR also increased the mRNA levels of these antioxidant enzymes upon exposure to oxidative stress. Rotifers cultured under CR showed a higher survival rate than those fed ad libitum (AL) upon exposure to 0.05-0.2 lM juglone, an oxidative stress inducer. The relative mRNA levels of Mn SOD and catalase before exposure to juglone were slightly higher in the CR rotifers than in their AL counterparts, although these differences were not statistically significant. AL rotifers showed no apparent upregulation of the mRNA levels of these antioxidant enzymes upon exposure to 0.025 and 0.05 lM juglone. In contrast, the CR rotifers increased the mRNA levels of Mn SOD and catalase by up to 5.4-fold and 4.2-fold, respectively, resulting in significant differences between their levels in AL and CR rotifers under oxidative stress conditions. Furthermore, the protein level of catalase was clearly higher in CR than in AL rotifers 6 h after exposure to oxidative stress. These results suggest that the upregulation of Mn SOD and catalase genes is involved in CR-induced resistance to oxidative stress in the rotifer.
A monogonont rotifer Brachionus plicatilis has been widely used as a model organism for physiological, ecological studies and for ecotoxicology. Because of the availability of parthenogenetic mode of reproduction as well as its versatility to be used as live food in aquaculture, the population dynamic studies using the rotifer have become more important and acquired the priority over those using other species. Although many studies have been conducted to identify environmental factors that influence rotifer populations, the molecular mechanisms involved still remain to be elucidated. In this study, gene(s) differentially expressed by calorie restriction in the rotifer was analyzed, where a calorie-restricted group was fed 3 h day(-1) and a well-fed group fed ad libitum. A subtracted cDNA library from the calorie-restricted rotifer was constructed using suppression subtractive hybridization (SSH). One hundred sixty-three expressed sequence tags (ESTs) were identified, which included 109 putative genes with a high identity to known genes in the publicly available database as well as 54 unknown ESTs. After assembling, a total of 38 different genes were obtained among 109 ESTs. Further validation of expression by semi-quantitative reverse transcription-PCR showed that 29 out of the 38 genes obtained by SSH were up regulated by calorie restriction.
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