Yanase, Sumino et al. (2009) International Worm Meeting "SOD-1 Deletions in Caenorhabditis elegans Alter the Localization of Intracellular ROS and Show Molecular Compensation."

E. Johnson, Thomas, Tedesco, Patricia, Onodera, Akira, Yanase, Sumino, Ishii, Naoaki

[International Worm Meeting, 2009]

Superoxide dismutase (SOD) is an enzyme that catalytically removes the superoxide radical (.O2-) and protects organisms from oxidative damage during normal aging. We demonstrate that not only the cytosolic .O2- level but also the mitochondrial .O2- level increased in the deletion mutants of sod-1 gene encoding Cu/Zn SOD in Caenorhabditis elegans (C. elegans). Interestingly, this suggests that the activity of SOD-1, which so far has been thought to act mainly in cytoplasm, helps to control the detoxification of .O2- also in the mitochondria. We also found functional compensation by other SODs, especially the sod-5 gene, which was induced several fold in the mutants. Therefore, the possibility exists that the compensative expression of sod-5 gene in the sod-1 deficit is associated with the insulin/insulin-like growth factor-1 (Ins/IGF-1) signaling pathway, which regulates longevity and stress resistance of C. elegans, because the sod-5 gene may be a target of the pathway (1). Reference (1) Yanase, S. and Ishii, N. (2008) Hyperoxia exposure induced hormesis decreases mitochondrial superoxide radical levels via Ins/IGF-1 signaling pathway in a long-lived age-1 mutant of Caenorhabditis elegans. Journal of Radiation Research 49(3): 211-218.

Yanase, Sumino et al. (2021) International Worm Meeting "Recovery of Muscle Function Dependent on the Impaired Cell Death in a C. elegans Premature Aging Model"

Suzuki, Yu, Yasuda, Kayo, Ishii, Naoaki, Yanase, Sumino, Sekigawa, Kota, Otsuka, Yuko

[International Worm Meeting, 2021]

In mammals including humans, the age-related losses of muscle mass and function are known as sarcopenia. Although the mechanisms underlying the development of sarcopenia are multifactorial, apoptosis has been shown to play a role in muscle loss during aging [1]. Recently, age-related muscle loss has also been found in the nematode C. elegans during normal aging. As in humans, age-related muscle loss in C. elegans leads to decreased mobility and serves as a marker for increased mortality [2, 3]. Here we examined the difference in age-associated pharyngeal muscle function between the short-lived mev-1(kn1) and the phenotype of deficient cell death (mev-1;ced-3 mutant), however, have shown no significant histopathological change in the loss of pharyngeal muscle mass. The mev-1 gene encodes a large subunit of the enzyme succinate dehydrogenase cytochrome b, which is a component of complex II in the mitochondrial electron transport chain. Mutation of the gene causes an increase in mitochondrial oxidative stress, thereby induces abnormal apoptosis in embryonic development and shortenes the lifespan [4]. Moreover, we aim to detect age-related difference in gene expression concerned with the cell death signaling using both mutants. References 1. Always SE et al. (2011) Aging and apoptosis in muscle, In: Handbook of the biology of aging, 7th edition (eds. Masoro EJ, Austad SN), Academic Press, London, pp. 63-118 2. Herndon LA et al. (2002) Stochastic and genetic factors influence tissue-specific decline in ageing C. elegans, Nature 419: 808-814 3. Fishen AL (2004) Of worms and women: Sarcopenia and its role in disability and mortality, J Am Geriatr Soc 52: 1185-1190 4. Ishii N et al. (1998) A mutation in succinate dehydrogenase cytochrome b causes oxidative stress and ageing in nematodes. Nature 394: 694-697

Yanase, Sumino et al. (2013) International Worm Meeting "Intermittent hyperoxia-induced hormesis decreases aerobic respiration via ins/IGF-1 and p53/CEP-1 signalings in C. elegans."

Yanase, Sumino, Shoyama, Tetsuji, Ishii, Naoaki, Suda, Hitoshi

[International Worm Meeting, 2013]

When a PI3-kinase mutant age-1, which has a long lifespan in C. elegans, was treated with intermittent hyperoxia, it extends the lifespan by the hormetic effect. We have previously reported that the hormetic lifespan extension is related to not only the inducing of antioxidant genes but also the decreasing of mitochondrial superoxide radical level via an ins/IGF-1 signaling pathway. In general, strains reduced the ins/IGF-1 signaling, such as age-1 and daf-2, have an elevated stress resistance, lower respiration rate, and an increased ATP concentration. It is suggested that intracellular metabolic shift to glycolysis from mitochondrial aerobic respiration occurs in the individual worm of the mutants compared to wild-type animals. We measured a change in the oxygen consumption level of the long-lived age-1 mutant in the hyperoxia-induced hormesis. Oxygen consumption level in age-1 animals significantly decreased after the intermittent hyperoxia exposure. In addition, expression of an antioxidant gst-4 gene, which is regulated by a homologue CEP-1 in C. elegans of a mammalian tumor suppressor p53, decreased in the hormetic age-1 mutant. Therefore, activity of p53/CEP-1, which is associated with the regulation of intracellular metabolic balance, was nontranscriptionally decreased because the induction of the p53/CEP-1 genes expresseion was not observed in the hormetic age-1 mutant.. Here, we described that the hyperoxia-induced hormetic lifespan extension in age-1 animals may be dependent on the metabolic shift to glycolysis from mitochondrial aerobic respiration via not only the ins/IGF-1 but also p53/CEP-1 signalings.

Sumino Yanase et al. (2003) International Worm Meeting "DAF-16 regulates anti-oxidant activities."

Sumino Yanase, Hiroko Uesugi, Naoaki Ishii, Yuji KOHARA

[International Worm Meeting, 2003]

The daf-16 gene encodes a forkhead transcription factor that presumably regulates the downstream target genes in a insulin-like signal pathway including DAF-12, which encodes an insulin receptor. It is still unknown what genes are regulated by DAF-16, but at least some genes related anti-oxidation are expected, because daf-2 mutants are resistant to oxidative stress and a daf-16 mutation suppresses this phenotype. The daf-16 mutant itself is also sensitive to oxygen. We examined what anti-oxidant enzymes are regulated by DAF-16. Furthermore, we analyzed a transcriptional profiling of daf-16(m26) allele using an expressed sequence tag (EST) microarray experiments to detect the target genes of DAF-16. As an experiment, we examined a Western blot analysis to determine the expression levels using antibodies to superoxide dismutases (SODs) and catalase in crude extract of daf-16(m26) mutants. In addition, the activities of SOD and catalase in the mutants were measured. We observed a reduction in Mn-SOD expression. Furthermore, the activities of Cu/Zn-SOD and catalase in the cytosol fraction were also reduced. Conversely, in the mev-1(kn1) mutant (which is sensitive to oxygen) as well as the daf-16(m26) mutants, reduction in gene expression and activities of these anti-oxidant enzymes were not observed. Using the microarray assay, the change of the expression level of a few anti-oxidant genes was found in daf-16(m26) mutants, e.g., the levels of a microsomal catalase (ctl-2) and a metallothionein gene (mtl-1) were decreased, and some major sperm protein genes (msp) were increased. These suggest that anti-oxidant proteins are strong candidates in the downstream target genes regulated by the DAF-16 transcription factor.

Sumino Yanase et al. (2004) East Asia Worm Meeting "Characterization of Phenotypes for Aging in Cu/Zn SOD Deletion Mutants"

Akira Onodera, Naoaki Ishii, Sumino Yanase

[East Asia Worm Meeting, 2004]

It is recently thought that the aging and longevity of life are controlled by the energy metabolism through an insulin-like signal transduction in the lower to higher organisms. Furthermore, it is speculated that the reactive oxygen species (ROS), the by-product on the route of the energy metabolic systems, are responsible for the aging process. It is also believed that the control of ROS production and protection of organic molecule from the produced ROS are somewhat connected to the lifespan extension. Superoxide dismutase (SOD) has a catalytic activity as the reduction of superoxide radicals. In SOD genes of Caenorhabditis elegans , sod-1 gene encoding a Cu/Zn SOD is mainly expressed in cytoplasm as a subcellular location. This gene is related to aging and acts to resist to oxidative damage. We have decided to investigate the phenotypes in aging, concentrating on 2 deletion mutants of sod-1 gene derived from National Bioresource Project. One of these deletion mutants, tm783, has its part of promoter regions, exon 1 and 2 deleted. The other strain, tm776, has its exon 3, 4 and 5 completely deleted. However, the average life span of these strains was slightly decreased compared to the one of wild type N2. We then measured sensitivities of these strains against oxidative stress using agents such as paraquat and oxygen. The oxygen resistances in tm776 and tm783 animals were not very clearly reduced than wild type. In contrast, the paraquat resistances in these strains were markedly decreased compared with wild type. In the sod-1 mutations, the tm783 strains showed a marked sensitivity to paraquat than tm776. It is likely that paraquat produces more ROS in cell at a single time than oxygen. Therefore, it is thought that result of the paraquat sensitivity test could reflect that there are much more activities within the intracellular anti-oxidant systems. tm783 animals may have decreased amount of sod-1 transcript, as this strain has the deletion in promoter region. Conversely, we could hypothesize that as the transcript of tm776 animals does not have the latter part the Cu/Zn SOD activities in tm776 animals are decreased. Thus, it is speculated that control of ROS is extremely important to extend the longevity of organisms such as the regulation of Cu/Zn SOD activities.

Sumino Yanase et al. (2005) International Worm Meeting "Characterization of Phenotypes for Aging in Cu/Zn SOD Deletion Mutants."

Akira Onodera, Naoaki Ishii, Sumino Yanase

[International Worm Meeting, 2005]

It is recently thought that the aging and longevity of life are controlled by the energy metabolism through an insulin-like signal transduction in the lower to higher organisms. Furthermore, it is speculated that the reactive oxygen species (ROS), the by-product on the route of the energy metabolic systems, are responsible for the aging process. It is also believed that the control of ROS production and protection of organic molecule from the produced ROS are somewhat connected to the lifespan extension. Superoxide dismutase (SOD) has a catalytic activity as the reduction of superoxide radicals. In four SOD genes of Caenorhabditis elegans (C. elegans), sod-1 gene encoding a Cu/Zn SOD is mainly expressed in cytoplasm as a subcellular location. This gene is related to aging and acts to resist to oxidative damage. We have decided to investigate the phenotypes in aging, concentrating on two deletion mutants of sod-1 gene derived from National Bioresource Project for the Experimental Animal Nematode C. elegans. One of these deletion mutants, tm783, has its part of promoter regions, exon 1 and 2 deleted. The other strain, tm776, has its exon 3, 4 and 5 completely deleted. Both strains are viable as homozygous. However, the average life span of these strains was slightly decreased compared to the one of wild type N2. We then measured sensitivities of these strains against oxidative stress using agents such as paraquat and oxygen. The oxygen resistances in tm776 and tm783 animals were not very clearly reduced than wild type. In contrast, the paraquat resistances in these strains were markedly decreased compared with wild type. In the sod-1 mutations, the tm783 strains showed a marked sensitivity to paraquat than tm776. It is likely that paraquat produces more ROS in cell at a single time than oxygen. Therefore, it is thought that result of the paraquat sensitivity test could reflect that there are much more activities within the intracellular anti-oxidant systems. tm783 animals may have decreased amount of sod-1 transcript, as this strain has the deletion in promoter region. Conversely, we could hypothesize that as the transcript of tm776 animals does not have the latter part the Cu/Zn SOD activities in tm776 animals are decreased. Thus, it is speculated that control of ROS is extremely important to extend the longevity of organisms such as the regulation of Cu/Zn SOD activities.

Sumino Yanase et al. (2008) C.elegans Aging, Stress, Pathogenesis, and Heterochrony Meeting "sod-5; A target gene of DAF-16 transcription factor in Caenorhabditis elegans"

Yoshihiro Takahashi, Sumino Yanase, Naohiro Suga, Naoaki Ishii

[C.elegans Aging, Stress, Pathogenesis, and Heterochrony Meeting, 2008]

We now report that a DAF-16 consensus binding element (DBE) sequence, which binds the DAF-16 transcription factor, was discovered in the promoter region of sod-5 gene encoding a Cu/Zn superoxide dismutase (SOD) in the nematode Caenorhabditis elegans (C. elegans). DAF-16 transcription factor is related to regulation of normal aging and oxidative stress resistance. The DBE position was 192 base pairs (bp) upstream of the start site for the transcription. In a daf-16 gene null mutant, daf-16(mgDf50) strain, the sod-5 gene expression as well as sod-3 gene was obviously suppressed compared with it in wild-type N2. In addition, the phenotype of lifespan in daf-16(mgDf50) animals was reduced when compared with N2 during normal aging.

Sumino Yanase et al. (2001) International C. elegans Meeting "An Adaptive Response Extends Life Span Through DAF-16 Transcription Factor in Caenorhabditis elegans."

Nao Ishii, Sumino Yanase

[International C. elegans Meeting, 2001]

Short-term exposure to high-concentration oxygen (90%) produces an adaptive response to oxidative stress in Caenorhabditis elegans . Thermotolerance, UV resistance and resistance against reactive oxygen species (ROS) are all consequences of this adaptive response. In this study, we found that life span of C. elegans was elongated by the adaptive response derived from short-term exposure to high-concentration oxygen as well as to heat shock stress. This extension was dependent on the stages and periods exposed with high-concentration oxygen. Moreover, this response was different in several mutants ( age-1 , mev-1 , daf-16 ) previously shown to have altered life spans. Accordingly, we speculated that the degree of the adaptive response plays an important role in determining life span. We measured mRNA levels of six genes ( sod-1 through sod-4 , ctl-1 , and ctl-2 ) known to be important in combating oxidative stress. Wild-type, age-1 , mev-1 and daf-16 animals were grown under atmospheric (21%) and 90% oxygen. There were considerable differences in the expression patterns of these six genes, most notably sod-1 , 2 and 3 levels were significantly reduced in daf-16 . This suggests that the adaptive response may mediate the relative intensity of resistance to oxidative stress, which was signaled to the daf-16 -dependent pathway. These data indicate that daf-16 must be functional in order to appropriately regulate the expression of these and other genes important in conferring resistance to oxidative stress. They further illustrate the importance of oxidative stress in the aging process.

Sumino Yanase et al. (2006) East Asia C. elegans Meeting "SOD-1 contributes to anti-aging by maintaining the balance of localization of intracellular ROS in Caenorhabditis elegans"

Sumino Yanase, Naoaki Ishii, Akira Onodera

[East Asia C. elegans Meeting, 2006]

It is recently thought that the aging and longevity of life are controlled by the energy metabolism through an insulin-like signal transduction in the lower to higher organisms. Furthermore, it is speculated that the reactive oxygen species (ROS), the by-product on the route of the energy metabolic systems, are responsible for the aging process. It is also believed that the control of ROS production and protection of organic molecule from the produced ROS are somewhat connected to the lifespan extension. Superoxide dismutase (SOD) has a catalytic activity as the reduction of superoxide radicals (O<SUB>2</SUB><SUP>-</SUP>). In five SOD genes of Caenorhabditis elegans (C. elegans), sod-1 gene encoding a Cu/Zn SOD is mainly expressed in cytoplasm as a subcellular location. This gene is related to aging and acts to resist to oxidative damage. We have decided to investigate the phenotypes in aging, concentrating on two deletion mutants of sod-1 gene derived from the National Bioresource Project for the Experimental Animal ""Nematode C. elegans"". One of these mutants, tm783, has its part of promoter regions, exon 1 and 2 deleted. The other strain, tm776, has its exon 3, 4 and 5 completely deleted. We measured sensitivities of these strains against oxidative stress using agents such as paraquat and oxygen. The oxygen resistances in tm776 and tm783 animals were not very clearly reduced than wild type. In contrast, the paraquat resistances in these strains were markedly decreased compared to wild type. The lifespans of both of the mutants were shortened approximately 10-20% compared to wild type. In addition, we demonstrate that both the mitochondrial and cytosol O<SUB>2</SUB><SUP>-</SUP> levels increased in the two mutants compared to wild type. This suggests that the activity of SOD-1, which is mainly cytoplasmic may help to control the ROS in the cytosol and mitochondria. The shortening of the lifespans in the deletion mutants seems to reflect both the influence on the lifespan by functional prevention of the sod-1 gene and functional substitution of other induced SODs dependent on the mitochondrial dysfunction.

Sumino Yanase et al. (2002) Japanese Worm Meeting "Adaptive responses to oxidative damage in three mutants of Caenorhabditis elegans (age-1, mev-1 and daf-16) that affect life span"

Naoaki Ishii, Kayo Yasuda, Sumino Yanase

[Japanese Worm Meeting, 2002]

Oxidative damage shortens the life span of the nematode Caenorhabditis elegans ( C. elegans ), even in an age-1 mutant that is characterized by a long life and oxygen resistance. We found that daily short-term exposure (3 hours) to hyperoxia further extended the life span of age-1 , a phenomenon known as an adaptive response. age-1 also showed resistance to paraquat and heat. Acute hyperoxic treatment did not extend the life spans of wild type, daf-16 or mev-1 . daf-16 mutant had a slightly shorter life span compared to wild type and was sensitive to heat and paraquat. The daf-16 phenotype resembles that of mev-1 showing a short life and oxygen sensitivity. We measured mRNA levels of superoxide dismutase genes ( sod-1 through 4 ), catalase genes ( clt-1 and ctl-2 ), known to encode anti-oxidant enzymes, and found they were elevated in age-1 young adults. On the other hand, in daf-16 and mev-1 , the expression of sod-1 , sod-2 and sod-3 genes was lower rather than in wild type. Conversely, ctl-1 and ctl-2 genes expression was significantly elevated in daf-16 and mev-1 . This suggests that DAF-16, a forkhead/winged-helix transcription factor, whose expression is suppressed by AGE-1, phosphoinositide 3-kinase (PI3-kinase), regulates anti-oxidant genes as well as energy metabolism under atmospheric conditions. However, the level of gene expression of SOD and catalase was not elevated by short-term exposure to 90% oxygen in wild type, mev-1 , daf-16 and even age-1 . This suggests that SOD and catalase do not play a role in the adaptive response against oxidative stress under hyperoxia, at least under these experimental conditions.