R of mitochondria could account for a decrease of oxidative stress in Akt1+/?mice.(Fig. 4F, G). Conversely, activation of AKT-1 by RNAi targeting daf-18 led to a shorter Pleuromutilin lifespan (Fig. 4H). Inhibition of ribosomal biogenesis by RNAi decreased the mitochondrial DNA content and extended the lifespan of wild-type animals (Fig. 4I, J), while inhibition of mitochondrial function by RNAi also increased the lifespan of wild-type animals (Fig. 4K). Moreover, inhibition of ribosomal biogenesis normalized the shortened lifespan of nematodes treated with daf-18 RNAi (Fig. 4L), suggesting that the decrease of ribosomal biogenesis and mitochondrial function were critical for improving the longevity of Akt1+/?mice.DiscussionThe present study demonstrated that haploinsufficiency of Akt1 significantly prolongs the lifespan of mice. Akt1+/?mice showed a decrease of ribosomal biogenesis and the amounts of mitochondria, which was associated with a reduction of oxidative stress. It has been reported that the insulin/IGF1 signaling pathway and the TOR pathway both promote ribosomal biogenesis and protein synthesis, as well as regulating mitochondrial function [15,44]. We found that the phospho-Akt1 level increased with age in wild-type mice, presumably due to constitutive activation of growth signals by various stresses. Given the large amount of energy consumed by ribosomal biogenesis and protein synthesis, Akt1+/?mice may utilize fewer mitochondria by reducing ribosomal biogenesis and thus minimize the enhancement of oxidative stress with aging, which could account for their longer lifespan. Decreased protein synthesis could improve the fidelity of biogenesis and prevent the accumulation of mis-folded proteins, which might also contribute to longevity associated with Akt1 deficiency. Since oxidative stress and oxygen consumption were both lower in Akt1+/?mice than in their littermates, an increase of mitochondria and energy expenditure associated with enhanced protein synthesis may accelerate aging and promote the onset of age-associated diseases. An intriguing aspect of longevity pathways is that they function independently of the cells affected, since mutations of one cell type can affect the phenotype of whole organism. Neuronal cells and adipose tissue have been suggested to have a critical role in regulation of the lifespan by the insulin/IGF1 pathway in nematodes and flies [10,45]. For example, increasing the level of DAF-16 expression in one tissue can increase DAF-16 activity elsewhere, through feedback regulation of insulin gene expression [46]. In addition, in daf-16 (?; daf-2 (? double mutant nematodes (short-lived animals), overexpression of DAF-16 in one tissue can increase the lifespan [47]. Inhibition of the insulin/IGF1 pathway in the brain or adipose tissue has been shown to induce longevity in mice [14,48], suggesting that lifespan can be regulated cell nonautonomously in mammals. In the present study, we did not determine the tissue(s) responsible for longevity in Akt1+/?mice.Ribosomal Biogenesis and Mitochondrial Function in Human Cells and C. elegansTo investigate the role of AKT1 in ribosomal biogenesis and in total mitochondrial activity, we 10236-47-2 infected human endothelial cells with a retroviral vector encoding a dominant-negative form of AKT1. Consistent with the results obtained in Akt1+/?mice, inhibition of AKT1 activity led to a decrease of oxygen consumption and ribosomal biogenesis compared with mockinfected cells (Fig. 4A, B). Th.R of mitochondria could account for a decrease of oxidative stress in Akt1+/?mice.(Fig. 4F, G). Conversely, activation of AKT-1 by RNAi targeting daf-18 led to a shorter lifespan (Fig. 4H). Inhibition of ribosomal biogenesis by RNAi decreased the mitochondrial DNA content and extended the lifespan of wild-type animals (Fig. 4I, J), while inhibition of mitochondrial function by RNAi also increased the lifespan of wild-type animals (Fig. 4K). Moreover, inhibition of ribosomal biogenesis normalized the shortened lifespan of nematodes treated with daf-18 RNAi (Fig. 4L), suggesting that the decrease of ribosomal biogenesis and mitochondrial function were critical for improving the longevity of Akt1+/?mice.DiscussionThe present study demonstrated that haploinsufficiency of Akt1 significantly prolongs the lifespan of mice. Akt1+/?mice showed a decrease of ribosomal biogenesis and the amounts of mitochondria, which was associated with a reduction of oxidative stress. It has been reported that the insulin/IGF1 signaling pathway and the TOR pathway both promote ribosomal biogenesis and protein synthesis, as well as regulating mitochondrial function [15,44]. We found that the phospho-Akt1 level increased with age in wild-type mice, presumably due to constitutive activation of growth signals by various stresses. Given the large amount of energy consumed by ribosomal biogenesis and protein synthesis, Akt1+/?mice may utilize fewer mitochondria by reducing ribosomal biogenesis and thus minimize the enhancement of oxidative stress with aging, which could account for their longer lifespan. Decreased protein synthesis could improve the fidelity of biogenesis and prevent the accumulation of mis-folded proteins, which might also contribute to longevity associated with Akt1 deficiency. Since oxidative stress and oxygen consumption were both lower in Akt1+/?mice than in their littermates, an increase of mitochondria and energy expenditure associated with enhanced protein synthesis may accelerate aging and promote the onset of age-associated diseases. An intriguing aspect of longevity pathways is that they function independently of the cells affected, since mutations of one cell type can affect the phenotype of whole organism. Neuronal cells and adipose tissue have been suggested to have a critical role in regulation of the lifespan by the insulin/IGF1 pathway in nematodes and flies [10,45]. For example, increasing the level of DAF-16 expression in one tissue can increase DAF-16 activity elsewhere, through feedback regulation of insulin gene expression [46]. In addition, in daf-16 (?; daf-2 (? double mutant nematodes (short-lived animals), overexpression of DAF-16 in one tissue can increase the lifespan [47]. Inhibition of the insulin/IGF1 pathway in the brain or adipose tissue has been shown to induce longevity in mice [14,48], suggesting that lifespan can be regulated cell nonautonomously in mammals. In the present study, we did not determine the tissue(s) responsible for longevity in Akt1+/?mice.Ribosomal Biogenesis and Mitochondrial Function in Human Cells and C. elegansTo investigate the role of AKT1 in ribosomal biogenesis and in total mitochondrial activity, we infected human endothelial cells with a retroviral vector encoding a dominant-negative form of AKT1. Consistent with the results obtained in Akt1+/?mice, inhibition of AKT1 activity led to a decrease of oxygen consumption and ribosomal biogenesis compared with mockinfected cells (Fig. 4A, B). Th.
