Effects of assay and acclimation temperatures on incorporation of amino acids into protein of isolated hepatocytes from the european eel, L.

Jankowsky, Hans-Dieter; Hotopp, W.; Vsiansky, P.

doi:10.1016/0306-4565(81)90007-3

Cited by 15 publications

(4 citation statements)

References 17 publications

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“…In an optimal bioassay, the exposure temperature should be chosen to balance the environmental relevance, optimal conditions for determining the biomarker or effect endpoint measured and offer a high-throughput system. In general, protein synthesis increases with temperature in fish (Jankowsky et al, 1981), and higher level of Vtg mRNA have been observed in rainbow trout hepatocytes exposed at 18C than at 14C (Pawlowski et al, 2000). This is similar to the present study where higher raw data readings (absorbance and fold change) was observed with increasing incubation temperatures.…”

Section: Isolation and Exposure Of Arctic Char Hepatocytessupporting

confidence: 91%

Primary hepatocytes from Arctic char (Salvelinus alpinus) as a relevant Arctic in vitro model for screening contaminants and environmental extracts

2017

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Contaminants find their way to the Arctic through long-range atmospheric transport, transport via ocean currents, and through increased anthropogenic activity. Some of the typical pollutants reaching the Arctic (PAHs, PCBs) are known to induce cytochrome P450 1a (CYP1A) protein expression and ethoxyresorufin-O-deethylase (EROD) activity through the aryl hydrocarbon receptor (AhR). In addition, some endocrine disrupting chemicals (EDCs) such as estrogen mimics (xenoestrogens) have been documented in Arctic areas and they may interfere with natural sexual development and reproduction. In vitro assays that are capable of detecting effects of such pollutants, covering multiple endpoints, are generally based on mammalian or temperate species and there are currently no well-characterized cell-based in vitro assays for effect assessment from Arctic fish species. The present study aimed to develop a high-throughput and multi-endpoint in vitro assay from Arctic char (Salvelinus alpinus) to provide a non-animal (alternative) testing method for an ecologically relevant Arctic species. A method for isolation and exposure of primary hepatocytes from Arctic char for studying the toxic effects and mode of action (MoA) of pollutants was applied and validated. The multi-versatility of the bioassay was assessed by classical biomarker responses such as cell viability (membrane integrity and metabolic activity), phase I detoxification (CYP1A protein expression, EROD activity) and estrogen receptor (ER) mediated vitellogenin (Vtg) protein expression using a selection of model compounds, environmental pollutants and an environmental extract containing a complex mixture of pollutants. Primary hepatocytes from Arctic char were successfully isolated and culture conditions optimized to identify the most optimal assay conditions for covering multiple endpoints. The hepatocytes responded with concentration-dependent responses to all of the model compounds, most of the environmental pollutants and the environmental sample tested. The bioassay response and sensitivity of the hepatocytes from Arctic char differed slightly from closely related salmonid species, thus highlighting the need for developing in vitro assays relevant for Arctic species. The present multi-endpoint in vitro assay offer a highly versatile tool to screen potential effects of pollutants and complex samples relevant for Arctic exposure scenarios.

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Section: Isolation and Exposure Of Arctic Char Hepatocytessupporting

confidence: 91%

Primary hepatocytes from Arctic char (Salvelinus alpinus) as a relevant Arctic in vitro model for screening contaminants and environmental extracts

2017

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“…As seen with Vtg production in vivo (Korsgaard et al 1986), in vitro production seems to be greatly limited at low temperatures. Although protein synthesis in general increases with temperature in fish (Jankowsky et al 1981), an optimum was obtained at 128C for Vtg synthesis in the hepatocyte monolayer culture when exposed to E2. In comparison, optimum Vtg production in a rainbow trout hepatocyte culture was obtained at a considerable higher temperature (208C) using identical assay conditions (K.-E. Tollefsen, unpublished data).…”

Section: Discussionmentioning

confidence: 94%

Induction of vitellogenin synthesis in an Atlantic salmon (Salmo salar) hepatocyte culture: a sensitivein vitrobioassay for the oestrogenic and anti-oestrogenic activity of chemicals

2003

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A variety of organic compounds have been documented to bind to the oestrogen receptor and induce oestrogenic effects in different vertebrates. The presence of these environmental oestrogens or oestrogen mimics in the aquatic environment has been suspected of disrupting the normal endocrinology of wild populations of fish. In this study, induction of vitellogenin synthesis in primary hepatocytes from Atlantic salmon (Salmo salar) was optimized and validated as an oestrogenic in vitro bioassay using a sensitive capture vitellogenin enzyme-linked immunosorbent assay. After proper optimization (cell media supplements, cell density, temperature and exposure time), this assay gave a sensitive and reproducible response to both endogenous steroids (relative potency: 17beta-oestradiol>>oestriol>oestrone>17alpha-oestradiol) and a range of common oestrogen mimics (relative potency: ethynyloestradiol and diethylstilboestrol>>genistein and zearalenone>>bisphenol A and 4-t-octylphenol>4-n-nonylphenol and 2'-chloro,4-chloro-diphenyltrichloroethane (o,p'-DDT). However, the androgen testosterone and the putative oestrogen mimics dieldrin and toxaphene were not shown to be oestrogenic using this hepatocyte bioassay. Oestrogen-induced vitellogenin synthesis was efficiently inhibited by the anti-oestrogen ZM 189.154, suggesting that this bioassay may be used for testing both the oestrogenic and the anti-oestrogenic properties of chemicals.

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“…This is the case for gill and white muscle, and data in the literature also indicate that this may be a common feature. Significantly reduced activation energies of protein synthesis rates were found in eel hepatocytes upon cold acclimation (Jankowsky et al, 1981). Many enzymes of cold-adapted species display reduced activation enthalpies to counterbalance the expected decrease in reaction velocity during cooling (Hochachka and Somero, 1984;Marshall, 1997;D'Amico et al, 2002).…”

Section: Rna Translational Capacity and Rna Translational Efficiencymentioning

confidence: 98%

Temperature-dependent protein synthesis capacities in Antarctic and temperate (North Sea) fish (Zoarcidae)

Storch

Lannig

Pörtner

2005

Journal of Experimental Biology

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For an evaluation of effects of seasonal cold acclimation and evolutionary cold adaptation on protein synthesis capacity, the protein synthesis apparatus was isolated from the gills and white muscle of Antarctic eelpout Pachycara brachycephalum and North Sea eelpout Zoarces viviparus. Both species had been acclimated to 0°C (control) and 5°C (Antarctic) and 5°C and 10°C (North Sea control). The translational capacities of the protein synthesis machineries were determined in an optimised cell-free in vitro system. The results demonstrate that tissues from the polar zoarcid possess cold-adapted protein synthesis machineries, indicated by low activation energies and, especially, high RNA translational capacities at similar RNA:protein ratios when compared to temperate zoarcids at 10°C. When both species were brought to 5°C, the temperate species displayed cold compensated protein synthesis capacities caused by elevated RNA:protein ratios. Warm exposure (from 0 to 5°C) of the Antarctic zoarcid revealed a capacity for thermal acclimation indicated by a reduction in protein synthesis capacities associated with lower RNA:protein ratios.

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Effects of assay and acclimation temperatures on incorporation of amino acids into protein of isolated hepatocytes from the european eel, L.

Cited by 15 publications

References 17 publications

Primary hepatocytes from Arctic char (Salvelinus alpinus) as a relevant Arctic in vitro model for screening contaminants and environmental extracts

Primary hepatocytes from Arctic char (Salvelinus alpinus) as a relevant Arctic in vitro model for screening contaminants and environmental extracts

Induction of vitellogenin synthesis in an Atlantic salmon (Salmo salar) hepatocyte culture: a sensitivein vitrobioassay for the oestrogenic and anti-oestrogenic activity of chemicals

Temperature-dependent protein synthesis capacities in Antarctic and temperate (North Sea) fish (Zoarcidae)

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