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  1. Article: Enhancement of immune proteins expression in skin mucus of Japanese flounder Paralicthys olivaceus upon feeding a diet supplemented with high concentration of ascorbic acid

    Mori, Misato / Ito, Tasuku / Washio, Ryota / Shibasaki, Yasuhiro / Namba, Aki / Yabu, Takeshi / Iwazaki, Dai / Wada, Noriko / Anzai, Hirosi / Shiba, Hajime / Nakanishi, Teruyuki / Mano, Nobuhiro

    Fish & shellfish immunology. 2021 July, v. 114

    2021  

    Abstract: To search immune defense proteins in skin mucus of Japanese flounder fed with a diet containing high concentration of ascorbic acid, we carried out 2D-PAGE and compared the resolved pattern of proteins between control group that fed commercial diet and ... ...

    Abstract To search immune defense proteins in skin mucus of Japanese flounder fed with a diet containing high concentration of ascorbic acid, we carried out 2D-PAGE and compared the resolved pattern of proteins between control group that fed commercial diet and ascorbic acid supplemented group (AsA group) fed a diet supplemented with high concentration of ascorbic acid (2,000 mg/kg) for 7 days. The results revealed that there were many proteins exhibited distinct increase in AsA group. Among them, 6 regions that showed a dramatic elevation were chosen for protein identification using LC-MS/MS analysis and Mascot database search. Six proteins were identified, i.e. serotransferrin (Sero), transferrin (Trans), warm temperature acclimation-related 65 kDa protein (Wap65), complement component c3 (C3), hemoglobin beta-A chain (Hbß) and apolipoprotein A-1 (Apo). Quantitative RT-PCR analysis showed that the mRNA level of Hbß in epidermis of AsA group gave much higher increase (11.6 folds) than control group; the levels of Sero/Trans, Wap65, C3 and Apo showed no apparent difference between the two groups. The mRNA levels of wap65 and c3 in the liver and Apo in the kidney of AsA group exhibited significant increase in comparison to control group. In the case of secreted immunoglobulin M (IgM) and lysozyme (lyz), no difference of the mRNA levels of IgM in epidermis, gill, kidney, spleen and intestine, and lyz in epidermis, gill, spleen and intestine, was observed. The results of in situ hybridization confirmed the elevation of Hbß mRNA level in the epidermis tissue of AsA group. Our present study provided additional evidence showing the effectiveness of AsA in activating innate immune defense system in skin mucosal tissue of fish.
    Keywords Paralichthys olivaceus ; apolipoprotein A-I ; ascorbic acid ; complement ; databases ; diet ; gene expression ; hemoglobin ; hybridization ; immune response ; innate immunity ; intestines ; kidneys ; liver ; lysozyme ; mucus ; reverse transcriptase polymerase chain reaction ; shellfish ; spleen ; temperature ; transferrin
    Language English
    Dates of publication 2021-07
    Size p. 20-27.
    Publishing place Elsevier Ltd
    Document type Article
    Note NAL-AP-2-clean
    ZDB-ID 1067738-0
    ISSN 1095-9947 ; 1050-4648
    ISSN (online) 1095-9947
    ISSN 1050-4648
    DOI 10.1016/j.fsi.2021.04.009
    Database NAL-Catalogue (AGRICOLA)

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  2. Article ; Online: Enhancement of immune proteins expression in skin mucus of Japanese flounder Paralicthys olivaceus upon feeding a diet supplemented with high concentration of ascorbic acid.

    Mori, Misato / Ito, Tasuku / Washio, Ryota / Shibasaki, Yasuhiro / Namba, Aki / Yabu, Takeshi / Iwazaki, Dai / Wada, Noriko / Anzai, Hirosi / Shiba, Hajime / Nakanishi, Teruyuki / Mano, Nobuhiro

    Fish & shellfish immunology

    2021  Volume 114, Page(s) 20–27

    Abstract: To search immune defense proteins in skin mucus of Japanese flounder fed with a diet containing high concentration of ascorbic acid, we carried out 2D-PAGE and compared the resolved pattern of proteins between control group that fed commercial diet and ... ...

    Abstract To search immune defense proteins in skin mucus of Japanese flounder fed with a diet containing high concentration of ascorbic acid, we carried out 2D-PAGE and compared the resolved pattern of proteins between control group that fed commercial diet and ascorbic acid supplemented group (AsA group) fed a diet supplemented with high concentration of ascorbic acid (2,000 mg/kg) for 7 days. The results revealed that there were many proteins exhibited distinct increase in AsA group. Among them, 6 regions that showed a dramatic elevation were chosen for protein identification using LC-MS/MS analysis and Mascot database search. Six proteins were identified, i.e. serotransferrin (Sero), transferrin (Trans), warm temperature acclimation-related 65 kDa protein (Wap65), complement component c3 (C3), hemoglobin beta-A chain (Hbß) and apolipoprotein A-1 (Apo). Quantitative RT-PCR analysis showed that the mRNA level of Hbß in epidermis of AsA group gave much higher increase (11.6 folds) than control group; the levels of Sero/Trans, Wap65, C3 and Apo showed no apparent difference between the two groups. The mRNA levels of wap65 and c3 in the liver and Apo in the kidney of AsA group exhibited significant increase in comparison to control group. In the case of secreted immunoglobulin M (IgM) and lysozyme (lyz), no difference of the mRNA levels of IgM in epidermis, gill, kidney, spleen and intestine, and lyz in epidermis, gill, spleen and intestine, was observed. The results of in situ hybridization confirmed the elevation of Hbß mRNA level in the epidermis tissue of AsA group. Our present study provided additional evidence showing the effectiveness of AsA in activating innate immune defense system in skin mucosal tissue of fish.
    Language English
    Publishing date 2021-04-20
    Publishing country England
    Document type Journal Article
    ZDB-ID 1067738-0
    ISSN 1095-9947 ; 1050-4648
    ISSN (online) 1095-9947
    ISSN 1050-4648
    DOI 10.1016/j.fsi.2021.04.009
    Database MEDical Literature Analysis and Retrieval System OnLINE

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