Functional and Ultrastructural Cell Pathology Induced by Fuel Oil in Cultured Dolphin Renal Cells

Pfeiffer, Carl J.; Sharova, Lioudmila V.; Gray, L.B.

doi:10.1006/eesa.2000.1950

Cited by 10 publications

(3 citation statements)

References 46 publications

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“…In cultures of cetacean (dolphin) kidney epithelial cells PAH exposure caused DNA damage and inhibited cell growth and exposure to fuel oil caused apoptosis and altered transport and enzyme activities after high exposure levels (Carvan et al,1995;Pfeiffer et al,2000;Sweat et al,2001). However, there are no studies with endothelial cells cultured from cetacean tissues which describe the dose dependent responses to toxicant exposure.…”

Section: Introductionmentioning

confidence: 99%

Cytochrome P4501A is induced in endothelial cell lines from the kidney and lung of the bottlenose dolphin, Tursiops truncatus

et al. 2006

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Marine mammals respond to the presence of polycyclic and planar halogenated aromatic hydrocarbons (PAH or PHAH) with the induced expression in endothelium of cytochrome P4501A1, regulated through the aryl hydrocarbon receptor (AHR) transcription factor. Physiological responses in other animals, such as edema and inflammation indicate that the endothelium may be compromised by exposure to AHR agonists, which are ubiquitous in the marine environment. In other mammals and fish the cellular and molecular consequences of exposure to AHR agonists have been elucidated in cultured endothelial cells. We have cultured and characterized cetacean endothelial cells (EC) and used them in induction studies. Endothelial cells were cultured from the lung and kidney of the bottlenose dolphin Tursiops truncatus and exposed to the AHR agonists β-naphthoflavone (βNF) and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). βNF (1-3 µM) induced significant increases in CYP1A1(O-deethylation of 7-ethoxyresorufin to resorufin;EROD) activity to 3.6 and 0.92 pmol/mg/min in lung and kidney EC, respectively. TCDD was more potent than βNF, and more efficacious, with maximum induction of CYP1A1activity of 10.1 and 15.2 pmol/mg/min in lung and kidney EC at 3-10 nM TCDD. The differential response indicates that the lung and kidney endothelial cells in culture retain the ability to respond in a selective manner to specific stimuli. Both the molecular mechanisms of induction and the physiological consequences, especially in the vasculature, of toxicant exposure can be studied in this system.

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Section: Introductionmentioning

confidence: 99%

Cytochrome P4501A is induced in endothelial cell lines from the kidney and lung of the bottlenose dolphin, Tursiops truncatus

et al. 2006

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“…Cytogenic evaluations using fibroblasts and kidney cells have been utilized in a number of different cetacean species such as beluga whale (Jarrell and Arnason, 1981) and the BND (Bielec et al, 1997, 1998). Cultured cells from various cetaceans have previously proven useful in assessing susceptibility to viruses (Cecil and Nigrelli, 1970; Nielsen et al, 1989; Kadoi et al, 1992; Dotzauer et al, 1994) and toxicants (DeGuise et al, 1998; Pfeiffer et al, 2000; Wang and Pfeiffer, 2001; Hu et al, 2002). Recent molecular studies performed on DK1/CDK bottlenose dolphin kidney cells demonstrate the value of cell cultures for testing relevant environmental agents such as halogenated aromatic hydrocarbons and the polynuclear aromatic hydrocarbons (Carvan et al, 2002).…”

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confidence: 99%

Establishment of epidermal cell lines derived from the skin of the Atlantic bottlenose dolphin (Tursiops truncatus)

Kindy

Ellis

et al. 2005

Anat. Rec.

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The Atlantic bottlenose dolphin (Tursiops truncatus), a marine mammal found off the Atlantic coast, has become the focus of considerable attention because of an increasing number of mortality events witnessed in this species over the last several years along the southeastern United States. Assessment of the impact of environmental stressors on bottlenose dolphins (BND) has been difficult because of the protected status of these marine mammals. The studies presented herein focused on establishing epidermal cell cultures and cell lines as tools for the in vitro evaluation of environmental stressors on BND skin. Epidermal cell cultures were established from skin samples obtained from Atlantic BND and subjected to karyotype analysis. These cultures were further characterized using immunohistochemical methods demonstrating expression of cytokeratins. By two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), we observed that the proteomic profile of BND skin tissue samples shared distinct similarities with that of skin-derived cultures. Epidermal cell cultures were transfected with a plasmid encoding the SV40 small t-and large T-antigens, as well as the neomycin-resistance gene. Five neomycin-resistant clones were isolated and expanded, and all of them proliferated at a faster rate than nontransfected BND epidermal cultures, which exhibited signs of senescence. Cell lysates prepared from two transfected clones were shown to express, by Western blot analysis, both SV40 tumor antigens. These experimental results are consistent with the concept that transfected clones expressing SV40 tumor antigens represent immortalized BND cell lines. Epidermal cell lines derived from Tursiops truncatus will provide a unique tool for studying key features of the interaction occurring between dolphins and the environment in which they live at their most crucial interface: the skin.

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“…Marine mammal cell culturing systems are considered rare multifunctional instruments for acquiring knowledge about the cell physiology and biochemistry of these animals as well as on the damaging effects of anthropogenic and natural toxicants [ 4 , 5 ]. Several cetacean cell types from multiple tissues have been isolated, including dermal fibroblasts [ 6 – 10 ], bronchial and muscle cells [ 4 ], alveolar macrophages [ 11 ], primary epithelial cells [ 12 ], kidney cells [ 13 , 14 ], blood cells [ 15 ] and cells from mesentery, lung, heart, liver, brain, spleen, thyroid, urinary bladder, periorbital soft tissue, and testes [ 16 ]. However, the use of cetacean primary cells for in vitro studies have been jeopardized due to their limited doubling capacity and their short life span.…”

Section: Introductionmentioning

confidence: 99%

Bottlenose dolphin (Tursiops truncatus) immortalized fibroblasts on novel 3D in vitro collagen-free scaffolds

Ferretti,

Moccia,

Centelleghe

et al. 2024

PLoS ONE

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Dolphins, as apex predators, can be considered relevant sentinels of the health of marine ecosystems. The creation of 3D cell models to assess in vitro cell-to-cell and cell-to-matrix interactions in environmental-mimicking conditions, is of considerable interest. However, to date the establishment of cetacean 3D culture systems has not yet been accomplished. Thus, in this study, different 3D systems of bottlenose dolphin (Tursiops truncatus) skin fibroblasts have been analyzed. Particularly, novel scaffolds based on hyaluronic acid and ionic-complementary self-assembling peptides such as RGD-EAbuK and EAbuK-IKVAV have been compared to Matrigel. Histological and fluorescent staining, electron microscopy (TEM) analyses and viability assays have been performed and RT-PCR has been used to detect extracellular matrix (ECM) components produced by cells. Results showed that Matrigel induced cells to form aggregates with lower viability and no ECM production compared to the novel scaffolds. Moreover, scaffolds allowed dispersed cells to produce a collagenous ECM containing collagen1a1, laminin B1 and elastin. The HA-EAbuK-IKVAV scaffold resulted in the most suitable 3D model in terms of cell quantity and viability. The development of this innovative approach is the first step towards the possibility to create 3D in vitro models for this protected species.

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Functional and Ultrastructural Cell Pathology Induced by Fuel Oil in Cultured Dolphin Renal Cells

Cited by 10 publications

References 46 publications

Cytochrome P4501A is induced in endothelial cell lines from the kidney and lung of the bottlenose dolphin, Tursiops truncatus

Cytochrome P4501A is induced in endothelial cell lines from the kidney and lung of the bottlenose dolphin, Tursiops truncatus

Establishment of epidermal cell lines derived from the skin of the Atlantic bottlenose dolphin (Tursiops truncatus)

Bottlenose dolphin (Tursiops truncatus) immortalized fibroblasts on novel 3D in vitro collagen-free scaffolds

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