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Dietary Lysine Imbalance Affects Muscle Proteome in Zebrafish (Danio rerio): A Comparative 2D-DIGE Study

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Abstract

Lysine (Lys) is an indispensable amino acid (AA) and generally the first limiting AA in vegetable protein sources in fish feeds. Inadequate dietary Lys availability may limit protein synthesis, accretion and growth of fish. This experiment aimed to further elucidate the role of Lys imbalance on growth by examining the myotomal muscle proteome of juvenile zebrafish (Danio rerio). Quadruplicate groups of 8 fish were fed either a low-Lys [Lys(−), 1.34 g kg−1], medium/control (Lys, 2.47 g kg−1) or high-Lys [Lys(+), 4.63 g kg−1] diet. Fish growth was monitored from 33 to 49 days post-fertilization (dpf) and trunk myotomal muscle proteome of Lys(−) and Lys(+) treatments were screened by 2D-DIGE and MALDI ToF tandem mass spectrometry. Growth rate was negatively affected by diet Lys(−). Out of 527 ± 11 (mean ± S.E.M.) protein spots detected (∼10–150 kDa and 4–7 pI value), 30 were over-expressed and 22 under-expressed in Lys(−) fish (|fold-change| >1.2, p value <0.05). Higher myosin light chains abundance and other myofibrillar proteins in Lys(−) fish pointed to increased sarcomeric degradation, indicating a higher protein turnover for supplying basal energy-saving metabolism rather than growth and muscle protein accretion. The Lys deficiency also possibly induced a higher feeding activity, reflected in the over-expression of beta enolase and mitochondrial ATP synthase. Contrarily, in the faster growing fish [Lys(+)], over-expression of apolipoprotein A-I, F-actin capping protein and Pdlim7 point to increased energy storage as fat and enhanced muscle growth, particularly by mosaic hyperplasia. Thus using an exploratory approach, this study pinpoints interesting candidates for further elucidating the role of dietary Lys on growth of juvenile fish.

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Acknowledgements

This research was funded by the Research Council of Norway (165203/S40) to IR. PGR was a recipient of a research contract from the University of Bergen. Support to the Proteomics lab was provided by the Meltzer Foundation, University of Bergen (KEF). LC and MdV participated with support of project HYDRAA-PTDC/MAR/71685/2006 (Fundação para a Ciência e Tecnologia (FCT), Portugal, with the support of FEDER). MdV was also supported by a grant SFRH/BD/40698/2007 (FCT, Portugal). Authors are especially grateful to Heikki Savolainen for excellent rearing of the fish and Ann Kristin Frøyset, Shailesh Narawane, Tomé S. Silva and Odete Cordeiro for useful technical assistance.

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Correspondence to Mahaut de Vareilles.

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de Vareilles, M., Conceição, L.E.C., Gómez-Requeni, P. et al. Dietary Lysine Imbalance Affects Muscle Proteome in Zebrafish (Danio rerio): A Comparative 2D-DIGE Study. Mar Biotechnol 14, 643–654 (2012). https://doi.org/10.1007/s10126-012-9462-3

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