The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Ĭompeting interests: The authors have declared that no competing interests exist. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.įunding: This study was financed by the Norwegian Research Council (project number NFR 185177/S40) and the SmartMix Programme of the Netherlands Ministry of Economic Affairs and the Netherlands Ministry of Education, Culture and Science. Received: NovemAccepted: JanuPublished: February 19, 2014Ĭopyright: © 2014 de Vrieze et al. PLoS ONE 9(2):Įditor: Christoph Winkler, National University of Singapore, Singapore The zebrafish scale is an excellent model to study how and which fatty acids affect skeletal formation.Ĭitation: de Vrieze E, Moren M, Metz JR, Flik G, Lie KK (2014) Arachidonic Acid Enhances Turnover of the Dermal Skeleton: Studies on Zebrafish Scales. The gene expression profile further suggests that arachidonic acid increases maturation rate of the regenerating scale in other words, enhances turnover. Arachidonic acid also clearly stimulates matrix metalloproteinase activity in vitro, which implies that resorptive effects of arachidonic acid are mediated by matrix metalloproteinases. Arachidonic acid affected scale regeneration (following removal of ontogenetic scales): mineral deposition was altered and both gene expression and enzymatic matrix metalloproteinase activity changed towards enhanced osteoclastic activity. The fatty acid profile resulting from a high arachidonic acid diet induced mild but significant increase in matrix resorption in ontogenetic scales of adult zebrafish. Elasmoid scales, dermal skeletal plates, are ideal to study skeletal metabolism in zebrafish and were exploited in the present study. To elucidate effects of poly-unsaturated fatty acids on developing and existing skeletal tissues, zebrafish ( Danio rerio) were fed (micro-) diets low and high in arachidonic acid content. Arachidonic acid is metabolized by cyclooxygenases to prostaglandin E2, an eicosanoid with effects on bone formation and remodeling. Supplementation of fish oils with vegetable oils increases the content of omega-6 fatty acids, such as arachidonic acid in the diet. With this model culture system, it is possible to evaluate the in vitro effects of hormones, growth factors, and other substances on growth and development of dermal skeleton in fish.In fish nutrition, the ratio between omega-3 and omega-6 poly-unsaturated fatty acids influences skeletal development. Our serum-free, chemically defined organ culture system enables in vitro development and growth of both scales and teeth. To evaluate our culture system, the medium that allowed to mimick in the best possible way the in vivo regeneration of scales (i.e., the basis medium plus dexamethasone and PG-E2) was also tested on the in vitro development of teeth in the same fish species. These effects were compared with those obtained by supplementation of the basis medium with newborn and fetal calf serum. With this basis medium, we examined the effects of all trans-retinoic acid, dexamethasone, and prostaglandin-E2 (PG-E2), factors known to exert an effect on development and growth of teeth and bone in mammalian culture systems, on the in vitro regeneration of scales. The culture medium in our system is based on Leibovitz medium (L-15) supplemented with vitamin C, additional amino acids, and HEPES. To develop a serum-free, chemically defined in vitro organ culture system enabling the study of epithelial-mesenchymal interactions in development and growth of fish dermal skeleton, we investigated in vitro continuation of scale regeneration in the cichlid fish Hemichromis bimaculatus.
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