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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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Related in: MedlinePlus

Northern blot analysis of B. mori SOD1 and SOD2.A. Total RNA (12 μg per lane) isolated from the B. mori fat body was analyzed by northern blot analysis using a probe for BmSOD1 and BmSOD2. A band at about 936 bases was identified as the BmSOD1 transcript (1), while a band at about 922 bases was identified as the BmSOD2 transcript (2). Total RNA was loaded for BmSOD1, BmSOD2 and lower panel of 18S rRNA (control of RNA loading).
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pone.0116007.g002: Northern blot analysis of B. mori SOD1 and SOD2.A. Total RNA (12 μg per lane) isolated from the B. mori fat body was analyzed by northern blot analysis using a probe for BmSOD1 and BmSOD2. A band at about 936 bases was identified as the BmSOD1 transcript (1), while a band at about 922 bases was identified as the BmSOD2 transcript (2). Total RNA was loaded for BmSOD1, BmSOD2 and lower panel of 18S rRNA (control of RNA loading).

Mentions: In the phylogenetic tree, vertebrate SOD1 and SOD2 and insect SOD1 and SOD2 were placed into four distinct clusters (Fig. 1-C). Northern blot analysis revealed that both transcription products were single products with characteristic sizes: 936 bases for BmSOD1 and 922 bases for BmSOD2 (Fig. 2).


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)

Northern blot analysis of B. mori SOD1 and SOD2.A. Total RNA (12 μg per lane) isolated from the B. mori fat body was analyzed by northern blot analysis using a probe for BmSOD1 and BmSOD2. A band at about 936 bases was identified as the BmSOD1 transcript (1), while a band at about 922 bases was identified as the BmSOD2 transcript (2). Total RNA was loaded for BmSOD1, BmSOD2 and lower panel of 18S rRNA (control of RNA loading).
© Copyright Policy
Related In: Results  -  Collection

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

pone.0116007.g002: Northern blot analysis of B. mori SOD1 and SOD2.A. Total RNA (12 μg per lane) isolated from the B. mori fat body was analyzed by northern blot analysis using a probe for BmSOD1 and BmSOD2. A band at about 936 bases was identified as the BmSOD1 transcript (1), while a band at about 922 bases was identified as the BmSOD2 transcript (2). Total RNA was loaded for BmSOD1, BmSOD2 and lower panel of 18S rRNA (control of RNA loading).
Mentions: In the phylogenetic tree, vertebrate SOD1 and SOD2 and insect SOD1 and SOD2 were placed into four distinct clusters (Fig. 1-C). Northern blot analysis revealed that both transcription products were single products with characteristic sizes: 936 bases for BmSOD1 and 922 bases for BmSOD2 (Fig. 2).

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