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The data herein is in support of our research article by McDougall et al. (2017) , in which we used our zebrafish model of embryonic vitamin E (VitE) deficiency to study the consequences of VitE deficiency during development. Adult 5D wild-type zebrafish (Danio rerio), fed defined diets without (E–) or with VitE (E+, 500 mg RRR-α-tocopheryl acetate/kg diet), were spawned to obtain E– and E+ embryos that we evaluated using metabolomics and specific lipid analyses (each measure at 24, 48, 72, 120 hours-post-fertilization, hpf), neurobehavioral development (locomotor responses at 96 hpf), and rescue strategies. Rescues were attempted using micro-injection into the yolksac using VitE (as a phospholipid emulsion containing d6-α-tocopherol at 0 hpf) or D-glucose (in saline at 24 hpf).
Value of the data
Fig. 1. shows data from quantitative analyses of LA (linoleic acid, 18:2, omega-6); ARA (arachidonic acid, 20:4, omega-6); EPA (eicosapentaenoic acid, 20:5, omega-3); DHA (docosahexaenoic acid, 22:6, omega-3) in fatty acid extracts from samples with and without alcoholic saponification of E– and E+ embryos collected at 24, 48, 72, and 120 hpf. Tables 1and 2 provide detailed targeted metabolomics datasets for E– and E+ embryos collected at 24, 48, 72, and 120 hpf. Relative response intensity metabolomics data for choline and methylation pathway intermediates in E– and E+ embryos are shown in Fig. 2. Relative response intensities of antioxidant network components from metabolomic analyses, as well as quantification of α-tocopherol and ascorbic acid, in E– and E+ embryos (pmol/embryo) are shown in Fig. 3. Relative response intensities of glycolytic and tricarboxylic acid cycle intermediates in E– and E+ embryos are shown in Fig. 4. Relative response intensities of free saturated fatty acids and coenzyme A from metabolomics data in E– and E+ embryo are shown in Fig. 5. Fig. 6 shows locomotor activity data from E– and E+ embryos micro-injected into the yolksac at 0 hpf with either saline or a VitE–emulsion. Fig. 7 shows locomotor activity data from E– and E+ embryos micro-injected into the yolksac at 24 hpf with either saline or D-glucose.
All experiments (i.e. lipid quantifications, targeted metabolomics analyses, and micro-injection rescue studies) were performed in duplicate and have been reported in detail .
The Institutional Animal Care and Use Committee of Oregon State University approved this protocol (ACUP Number: 4344). Tropical 5D strain zebrafish were housed in the Sinnhuber Aquatic Research Laboratory and complete details of the housing and husbandry have been reported .
Extraction and sample preparation for metabolomic analysis were performed following 24, 48, 72, and 120 hpf, embryos (n=15 per replicate, n=4 replicates per group), as described . Chromatography was performed with a Shimadzu Nexera system (Shimadzu; Columbia, MD, USA) coupled to a high-resolution hybrid quadrupole–time–of-flight mass spectrometer (TripleTOF® 5600; SCIEX; Framingham, MA, USA). Two different LC analyses using reverse phase and HILIC columns were used, as described .
Analysis of total DHA, EPA, ARA, and LA were performed as described  with modifications, as described . Chromatographic separations were carried out on 4.6×250 mm J׳sphere ODS-H80 (4 µm, YMC Co, Kyoto, Japan) for negative ion analysis. TOF-MS and TOF-MS/MS were operated with same parameters as for metabolomics, as described .
Embryos were microinjected as described and criteria used to assess supplementation tolerance of zebrafish embryos using ZAAP at 24, 48, and 120 hpf, as described .
The authors thank Carrie Barton, Greg Gonnerman, Andrea Knecht, Jane La Du, Scott Leonard, and Lisa Truong for providing outstanding technical assistance. National Institutes of Health Grants S10RR027878 (MGT and JFS) and NIEHS P30 ES000210 (RT) supported this work. MM was supported in part by National Science Foundation Grant DGE 0965820. H-KK sabbatical support provided by The Catholic University of Korea. MGT supported in part by the Helen P Rumbel endowment to the Linus Pauling Institute.