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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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Effects of ROT- and ISDN-induced oxidative stress on BmSOD1 and BmSOD2 expression in BmN4 cells.BmN4 cells exposed to ROT for 3 or 6 hours (A) or ISDN for 3 or 6 hours (B) were examined for BmSOD1 and BmSOD2 content by immunoblotting. Aliquots (10 μg) of protein samples from BmN4 cells subjected to the following treatments: control (lanes 1–3) and ROT treatment (lane 4–6) for 3 hours; control (lane 7–9) and ROT treatment (lane 10–12). All samples were separated by SDS-PAGE, transferred to nitrocellulose and probed with each specific antibody.
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pone.0116007.g006: Effects of ROT- and ISDN-induced oxidative stress on BmSOD1 and BmSOD2 expression in BmN4 cells.BmN4 cells exposed to ROT for 3 or 6 hours (A) or ISDN for 3 or 6 hours (B) were examined for BmSOD1 and BmSOD2 content by immunoblotting. Aliquots (10 μg) of protein samples from BmN4 cells subjected to the following treatments: control (lanes 1–3) and ROT treatment (lane 4–6) for 3 hours; control (lane 7–9) and ROT treatment (lane 10–12). All samples were separated by SDS-PAGE, transferred to nitrocellulose and probed with each specific antibody.

Mentions: Expression levels of both BmSOD proteins were the same for exposure to ROT for 3 and 6 hours (Fig. 6-A and S4 Fig.). With exposure to ISDN, a nitric oxide (NO) generator [23], expression levels of ISDN-treated BmN4 cells were the same as for the control (Fig. 6-B and S5 Fig.). Thus, ROT and ISDN did not affect the expression levels of either BmSOD protein (Fig. 6 A, B).


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)

Effects of ROT- and ISDN-induced oxidative stress on BmSOD1 and BmSOD2 expression in BmN4 cells.BmN4 cells exposed to ROT for 3 or 6 hours (A) or ISDN for 3 or 6 hours (B) were examined for BmSOD1 and BmSOD2 content by immunoblotting. Aliquots (10 μg) of protein samples from BmN4 cells subjected to the following treatments: control (lanes 1–3) and ROT treatment (lane 4–6) for 3 hours; control (lane 7–9) and ROT treatment (lane 10–12). All samples were separated by SDS-PAGE, transferred to nitrocellulose and probed with each specific antibody.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0116007.g006: Effects of ROT- and ISDN-induced oxidative stress on BmSOD1 and BmSOD2 expression in BmN4 cells.BmN4 cells exposed to ROT for 3 or 6 hours (A) or ISDN for 3 or 6 hours (B) were examined for BmSOD1 and BmSOD2 content by immunoblotting. Aliquots (10 μg) of protein samples from BmN4 cells subjected to the following treatments: control (lanes 1–3) and ROT treatment (lane 4–6) for 3 hours; control (lane 7–9) and ROT treatment (lane 10–12). All samples were separated by SDS-PAGE, transferred to nitrocellulose and probed with each specific antibody.
Mentions: Expression levels of both BmSOD proteins were the same for exposure to ROT for 3 and 6 hours (Fig. 6-A and S4 Fig.). With exposure to ISDN, a nitric oxide (NO) generator [23], expression levels of ISDN-treated BmN4 cells were the same as for the control (Fig. 6-B and S5 Fig.). Thus, ROT and ISDN did not affect the expression levels of either BmSOD protein (Fig. 6 A, B).

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