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Lumican is a major small leucine-rich proteoglycan (SLRP) in Atlantic cod (Gadus morhua L.) skeletal muscle

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Abstract

Knowledge on fish matrix biology is important to ensure optimal fish -quality, -growth and -health in aquaculture. The aquaculture industry face major challenges related to matrix biology, such as inflammations and malformations. Atlantic cod skeletal muscle was investigated for collagen I, decorin, biglycan, and lumican expression and distribution by real-time PCR, immunohistochemical staining and Western blotting. Immunohistochemical staining and Western immunoblotting were also performed using antibodies against glycosaminoglycan side chains of these proteoglycans, in addition to fibromodulin. Real-time PCR showed highest mRNA expression of lumican and collagen I. Collagen I and proteoglycan immunohistochemical staining revealed distinct thread-like structures in the myocommata, with the exception of fibromodulin, which stained in dense structures embedded in the myocommata. Chondroitinase AC-generated epitopes stained more limited than cABC-generated epitopes, indicating a stronger presence of dermatan sulfate than chondroitin sulfate in cod muscle. Lumican and keratan sulfate distribution patterns were strong and ubiquitous in endomysia and myocommata. Western blots revealed similar SLRPs sizes in cod as are known from mammals. Staining of chondroitin/dermatan sulfate epitopes in Western blots were similar in molecular size to those of decorin and biglycan, whereas staining of keratan sulfate epitopes coincided with expected molecular sizes of lumican and fibromodulin. In conclusion, lumican was a major proteoglycan in cod muscle with ubiquitous distribution overlapping with keratan sulfate. Other leucine-rich proteoglycans were also present in cod muscle, and Western blot using antibodies developed for mammalian species showed cross reactivity with fish, demonstrating similar structures and molecular weights as in mammals.

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Acknowledgements

This study was financed through the Norwegian Research Council grant nr. 185271/S40 and also supported by grants from the Throne Holst Foundation. Heidi Mikalsen at Nofima was valuable for organizing and helping during sampling. Dr. Sissel B. Rønning at Nofima was very helpful during establishment of immunprecipitation protocols. BioMar provided feed for the fish used in this study. We are very grateful to the Norwegian Cod Genome Sequencing Consortium for access to the Atlantic cod genome database (www.codgenome.no) in advance of publication.

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Authors and Affiliations

  1. The Norwegian Institute of Food, Fisheries and Aquaculture Research, Nofima AS, Osloveien 1, N-1430, Ås, Norway

    Monica G. Tingbø, Mona E. Pedersen, Grethe Enersen & Kirsten O. Hannesson

  2. Department of Nutrition, Faculty of Medicine, University of Oslo, Oslo, Norway

    Svein O. Kolset

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  1. Monica G. Tingbø
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  2. Mona E. Pedersen
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  3. Svein O. Kolset
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Correspondence to Monica G. Tingbø.

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Tingbø, M.G., Pedersen, M.E., Kolset, S.O. et al. Lumican is a major small leucine-rich proteoglycan (SLRP) in Atlantic cod (Gadus morhua L.) skeletal muscle. Glycoconj J 29, 13–23 (2012). https://doi.org/10.1007/s10719-011-9358-x

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