Zebrafish (Danio rerio) fed vitamin E-deficient diets produce embryos with increased morphologic abnormalities and mortality

Abstract

Vitamin E (α-tocopherol) is required to prevent fetal resorption in rodents. To study α–tocopherol’s role in fetal development, a non-placental model is required. Therefore, the zebrafish, an established developmental model organism, was studied by feeding the fish a defined diet with or without added α–tocopherol. Zebrafish (age: 4–6 w) were fed the deficient (E-), sufficient (E+), or lab diet up to 1 y. All groups showed similar growth rates. The exponential rate of α–tocopherol depletion up to ~80 day in E- zebrafish was 0.029 ± 0.006 nmol/g, equivalent to a depletion half-life of 25 ± 5 days. From age ~80 d, the E- fish (5 ± 3 nmol/g) contained ~50 times less α–tocopherol than the E+ or lab diet fish (369 ± 131 or 362 ± 107, respectively, P<0.05). E-depleted adults demonstrated decreased startle response suggesting neurologic deficits. Expression of selected oxidative stress and apoptosis genes from livers isolated from the zebrafish fed the three diets were evaluated by quantitative polymerase chain reaction and were not found to vary with vitamin E status. When E-depleted adults were spawned, they produced viable embryos with depleted α–tocopherol concentrations. The E- embryos exhibited a higher mortality (P<0.05) at 24 h post fertilization (hpf) and a higher combination of malformations and mortality (P<0.05) at 120 hpf than embryos from parents fed E+ or lab diets. This study documents for the first time that vitamin E is essential for normal zebrafish embryonic development.

Novel function of vitamin E in regulation of zebrafish (Danio rerio) brain lysophospholipids discovered using lipidomics

“…Adult zebrafi sh were kept under standard laboratory conditions at 28.5°C with a 14 h light/10 h dark cycle. For 9 months, beginning at 50 days postfertilization, zebrafi sh were fed defi ned diets, which contained only fatty acids shorter than 18 carbons with two or three double bonds ( 2,5,9 ), were prepared in 300 g batches with the vitamin C source as StayC (500 mg/kg; Argent Chemical Laboratories Inc.) without (E Ϫ ) or with (E+) added vitamin E (500 mg RRR -␣ -tocopheryl acetate/kg diet; ADM, Decatur, IL), as described previously ( 5 ). Diets were stored at Ϫ 20°C until fed to the zebrafi sh.…”

PEG-<em>b</em>-PCL polymeric nano-micelle inhibits vascular angiogenesis by activating p53-dependent apoptosis in zebrafish

“…A total of 10 μL reaction system was prepared for real-time quantitative polymerase chain reaction (RT-qPCR) using SYBR Green Master Mix (TaKaRa). The sequences of used primers were flk1 forward 5′-GAGAACGGAACCAACAAGATCCAC GAG-3′ reverse 5′-CCCTCCAGCAGAACTGACTCCTTAC-3′ p53 forward 5′-ACCACTGGGACCAAACGTAG-3′ reverse 5′-CAGAGTCGCTTCTTCCTTCG-3′, 32 AIF forward 5′-AAAGTCCGGAAAGAGGGTGT-3′ reverse 5′-GCCTGGAGCTCAGCATTAAC-3′, 33 β -actin forward 5′ -CTGTCTTCCCATCCATCG TGGGTC-3′ reverse 5′-CTCCATATCATCCCAGTTGGTGACA-3′.…”

Outcome of a public consultation on the Draft Scientific Opinion of the EFSA Panel on Dietetic Products, Nutrition and Allergies (NDA) on Dietary Reference Values for vitamin E as ɑ‐tocopherol

“…Comments received: 6. It was suggested that epidemiological studies may provide a better representation of the general population than randomised controlled trials (RCTs).…”

“…The importance of vitamin E in protecting polyunsaturated fatty acids (PUFAs) against oxidation in humans was emphasised. Research on the effects of vitamin E deficiency in mice (Tanito et al, 2007) and zebrafish (Danio reio) (Miller et al, 2012; Lebold et al, 2013) was referred to in order to support the importance of vitamin E in cellular membranes high in docosahexaenoic acid (DHA) and arachidonic acid (AA) in the brain, nervous system, retina, and reproductive system, and therefore development in early life.2. It was added that the vitamin E presence in cell membranes (preferentially in non-raft domains) is well documented .…”

“…The two publications by Horwitt mentioned in the comments received were already taken into account (Section 2.4.2. of the Scientific Opinion) and their usefulness for setting α-tocopherol requirement was assessed in the Scientific Opinion. The studies by Tanito et al (2007), Miller et al (2012 and Lebold et al (2013) describe data obtained using different animal models, but these data do not provide additional information for the setting of DRVs for α-tocopherol in humans.Ad2.In an in-vitro study, Howard et al (2011) The Panel considered that no change in the Scientific Opinion was needed in relation to these comments. …”