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Superoxide dismutases, SOD1 and SOD2, play a distinct role in the fat body during pupation in silkworm Bombyx mori.

Nojima Y, Ito K, Ono H, Nakazato T, Bono H, Yokoyama T, Sato R, Suetsugu Y, Nakamura Y, Yamamoto K, Satoh J, Tabunoki H, Fugo H - PLoS ONE (2015)

Bottom Line: One way that aerobic biological systems counteract the generation of reactive oxygen species (ROS) is with superoxide dismutase proteins SOD1 and SOD2 that metabolize superoxide radicals to molecular oxygen and hydrogen peroxide or scavenge oxygen radicals produced by the extensive oxidation-reduction and electron-transport reactions that occur in mitochondria.We found that BmSOD2 had a unique expression pattern in the fat body through the fifth instar larval developmental stage.These results suggest that BmSOD1 and BmSOD2 modulate environmental oxidative stress in the cell and have a specific role in fat body of B. mori during pupation.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Production, Faculty of Agriculture, Tokyo University of Agriculture and Technology, Saiwai-cho 3-5-8, Fuchu, Tokyo 183-8509, Japan; Department of Bioinformatics and Molecular Neuropathology, Meiji Pharmaceutical University, Noshio, 2-522-1, Kiyose, Tokyo 204-8588, Japan.

ABSTRACT
One way that aerobic biological systems counteract the generation of reactive oxygen species (ROS) is with superoxide dismutase proteins SOD1 and SOD2 that metabolize superoxide radicals to molecular oxygen and hydrogen peroxide or scavenge oxygen radicals produced by the extensive oxidation-reduction and electron-transport reactions that occur in mitochondria. We characterized SOD1 and SOD2 of Bombyx mori isolated from the fat body of larvae. Immunological analysis demonstrated the presence of BmSOD1 and BmSOD2 in the silk gland, midgut, fat body, Malpighian tubules, testis and ovary from larvae to adults. We found that BmSOD2 had a unique expression pattern in the fat body through the fifth instar larval developmental stage. The anti-oxidative functions of BmSOD1 and BmSOD2 were assessed by exposing larvae to insecticide rotenone or vasodilator isosorbide dinitrate, which is an ROS generator in BmN4 cells; however, exposure to these compounds had no effect on the expression levels of either BmSOD protein. Next, we investigated the physiological role of BmSOD1 and BmSOD2 under environmental oxidative stress, applied through whole-body UV irradiation and assayed using quantitative RT-PCR, immunoblotting and microarray analysis. The mRNA expression level of both BmSOD1 and BmSOD2 was markedly increased but protein expression level was increased only slightly. To examine the differences in mRNA and protein level due to UV irradiation intensity, we performed microarray analysis. Gene set enrichment analysis revealed that genes in the insulin signaling pathway and PPAR signaling pathway were significantly up-regulated after 6 and 12 hours of UV irradiation. Taken together, the activities of BmSOD1 and BmSOD2 may be related to the response to UV irradiation stress in B. mori. These results suggest that BmSOD1 and BmSOD2 modulate environmental oxidative stress in the cell and have a specific role in fat body of B. mori during pupation.

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Quantitative RT-PCR confirmation of BmSOD1 and BmSOD2 expression in UV-irradiated fat body.(A) BmSOD1, (B) BmSOD2. mRNA expression in fat bodies pooled from larvae subjected to UV-irradiation at 4.86 J/cm2 (n = 3), 9.72 J/cm2 (n = 3), 58.32 J/cm2 (n = 4) and non-irradiated control (n = 5) was plotted as RQ values. Error bars indicate the relative minimum/maximum expression levels against mean RQ expression levels. Technical replication was performed triplicate. Statistical significance was determined by Student’s t-test. *, P<0.05; **, P<0.01; and ***, P<0.001 compared with control values.
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pone.0116007.g007: Quantitative RT-PCR confirmation of BmSOD1 and BmSOD2 expression in UV-irradiated fat body.(A) BmSOD1, (B) BmSOD2. mRNA expression in fat bodies pooled from larvae subjected to UV-irradiation at 4.86 J/cm2 (n = 3), 9.72 J/cm2 (n = 3), 58.32 J/cm2 (n = 4) and non-irradiated control (n = 5) was plotted as RQ values. Error bars indicate the relative minimum/maximum expression levels against mean RQ expression levels. Technical replication was performed triplicate. Statistical significance was determined by Student’s t-test. *, P<0.05; **, P<0.01; and ***, P<0.001 compared with control values.

Mentions: Examination of expression levels of mRNA for both BmSOD by qRT-PCR after UV irradiation at 4.86, 9.72 and 58.32 J/cm2 showed that BmSOD1 and BmSOD2 mRNA expression was significantly increased in the 9.72 J/cm2 group (Fig. 7), while protein expression levels of BmSOD1 and BmSOD2 were slightly increased in the UV irradiation groups (Fig. 8 and S6 Fig.).


Superoxide dismutases, SOD1 and SOD2, play a distinct role in the fat body during pupation in silkworm Bombyx mori.

Nojima Y, Ito K, Ono H, Nakazato T, Bono H, Yokoyama T, Sato R, Suetsugu Y, Nakamura Y, Yamamoto K, Satoh J, Tabunoki H, Fugo H - PLoS ONE (2015)

Quantitative RT-PCR confirmation of BmSOD1 and BmSOD2 expression in UV-irradiated fat body.(A) BmSOD1, (B) BmSOD2. mRNA expression in fat bodies pooled from larvae subjected to UV-irradiation at 4.86 J/cm2 (n = 3), 9.72 J/cm2 (n = 3), 58.32 J/cm2 (n = 4) and non-irradiated control (n = 5) was plotted as RQ values. Error bars indicate the relative minimum/maximum expression levels against mean RQ expression levels. Technical replication was performed triplicate. Statistical significance was determined by Student’s t-test. *, P<0.05; **, P<0.01; and ***, P<0.001 compared with control values.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4340916&req=5

pone.0116007.g007: Quantitative RT-PCR confirmation of BmSOD1 and BmSOD2 expression in UV-irradiated fat body.(A) BmSOD1, (B) BmSOD2. mRNA expression in fat bodies pooled from larvae subjected to UV-irradiation at 4.86 J/cm2 (n = 3), 9.72 J/cm2 (n = 3), 58.32 J/cm2 (n = 4) and non-irradiated control (n = 5) was plotted as RQ values. Error bars indicate the relative minimum/maximum expression levels against mean RQ expression levels. Technical replication was performed triplicate. Statistical significance was determined by Student’s t-test. *, P<0.05; **, P<0.01; and ***, P<0.001 compared with control values.
Mentions: Examination of expression levels of mRNA for both BmSOD by qRT-PCR after UV irradiation at 4.86, 9.72 and 58.32 J/cm2 showed that BmSOD1 and BmSOD2 mRNA expression was significantly increased in the 9.72 J/cm2 group (Fig. 7), while protein expression levels of BmSOD1 and BmSOD2 were slightly increased in the UV irradiation groups (Fig. 8 and S6 Fig.).

Bottom Line: One way that aerobic biological systems counteract the generation of reactive oxygen species (ROS) is with superoxide dismutase proteins SOD1 and SOD2 that metabolize superoxide radicals to molecular oxygen and hydrogen peroxide or scavenge oxygen radicals produced by the extensive oxidation-reduction and electron-transport reactions that occur in mitochondria.We found that BmSOD2 had a unique expression pattern in the fat body through the fifth instar larval developmental stage.These results suggest that BmSOD1 and BmSOD2 modulate environmental oxidative stress in the cell and have a specific role in fat body of B. mori during pupation.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Production, Faculty of Agriculture, Tokyo University of Agriculture and Technology, Saiwai-cho 3-5-8, Fuchu, Tokyo 183-8509, Japan; Department of Bioinformatics and Molecular Neuropathology, Meiji Pharmaceutical University, Noshio, 2-522-1, Kiyose, Tokyo 204-8588, Japan.

ABSTRACT
One way that aerobic biological systems counteract the generation of reactive oxygen species (ROS) is with superoxide dismutase proteins SOD1 and SOD2 that metabolize superoxide radicals to molecular oxygen and hydrogen peroxide or scavenge oxygen radicals produced by the extensive oxidation-reduction and electron-transport reactions that occur in mitochondria. We characterized SOD1 and SOD2 of Bombyx mori isolated from the fat body of larvae. Immunological analysis demonstrated the presence of BmSOD1 and BmSOD2 in the silk gland, midgut, fat body, Malpighian tubules, testis and ovary from larvae to adults. We found that BmSOD2 had a unique expression pattern in the fat body through the fifth instar larval developmental stage. The anti-oxidative functions of BmSOD1 and BmSOD2 were assessed by exposing larvae to insecticide rotenone or vasodilator isosorbide dinitrate, which is an ROS generator in BmN4 cells; however, exposure to these compounds had no effect on the expression levels of either BmSOD protein. Next, we investigated the physiological role of BmSOD1 and BmSOD2 under environmental oxidative stress, applied through whole-body UV irradiation and assayed using quantitative RT-PCR, immunoblotting and microarray analysis. The mRNA expression level of both BmSOD1 and BmSOD2 was markedly increased but protein expression level was increased only slightly. To examine the differences in mRNA and protein level due to UV irradiation intensity, we performed microarray analysis. Gene set enrichment analysis revealed that genes in the insulin signaling pathway and PPAR signaling pathway were significantly up-regulated after 6 and 12 hours of UV irradiation. Taken together, the activities of BmSOD1 and BmSOD2 may be related to the response to UV irradiation stress in B. mori. These results suggest that BmSOD1 and BmSOD2 modulate environmental oxidative stress in the cell and have a specific role in fat body of B. mori during pupation.

Show MeSH
Related in: MedlinePlus