Isolation and culture of endocardial endothelial cells from Atlantic salmon (

Koren, Christian; Sveinbjørnsson, Baldur; Smedsrød, Bård

doi:10.1007/s004410050911

Cited by 24 publications

(16 citation statements)

References 31 publications

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“…A recently developed method (22) to isolate and culture cod atrial endocardial endothelial cells was used to show that these cells avidly take up fluorescently labeled bovine collagen alpha chains in vitro (Fig. 2a).…”

Section: Resultsmentioning

confidence: 99%

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Scavenger endothelial cells of vertebrates: A nonperipheral leukocyte system for high-capacity elimination of waste macromolecules

Seternes¹,

Sørensen²,

Smedsrød³

2002

Proc. Natl. Acad. Sci. U.S.A.

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131

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I n his pioneering work on the ultrastructure of the hepatic sinusoid 30 years ago, Wisse (1) observed numerous ''smoothwalled macropinocytic vesicles'' in the sinusoidal liver endothelial cells (LEC), suggestive of active pinocytic scavenging of blood plasma proteins. The subsequent finding that the specific activities of lysosomal enzymes in LEC were as high as in the Kupffer cells (2), and for some enzymes even higher, substantiated Wisse's notion. In a series of papers over the next 20 years, it was shown that LEC eliminate an array of soluble macromolecular physiologic and foreign waste products from the circulation by receptor-mediated endocytosis (3-7). It is now well established that the population of LEC in mammals serves as the most important site of elimination of an array of circulating soluble macromolecular waste products. To carry out this scavenger function, LEC express at least five types of specific receptors for endocytosis of major physiologic waste products: (i) the hyaluronan receptor for major matrix polysaccharides and proteoglycans such as hyaluronan and chondroitin sulfate (8); (ii) the collagen alpha-chain receptor for collagen alpha-chains of several types of collagen (9); (iii) the scavenger receptor for amino-terminal propeptides of types I and III procollagen (4), modified macromolecules such as atherogenic advanced glycation end (AGE) products (5) and oxidized LDL (6), oligodeoxynucleotides (10), and blood proteins modified by the process of blood clotting and platelet activation (11); (iv) the mannose receptor for carboxy-terminal propeptide of type I procollagen (7) and tissue plasminogen activator (12); and (v) the Fc-␥ receptor for IgG-antigen immune complexes (13). It is interesting to note that the LEC are able to clear all major categories of biological macromolecules (proteins, polysaccharides, lipids, and nucleic acids) by means of these five types of endocytosis receptors. The blood concentration of most of these substances is normally very low (ng͞ml) because of highly efficient elimination by endocytosis receptors on LEC.On the basis of several studies showing that LEC represent an important nonphagocytic scavenger cell in mammals, it has been suggested that the hepatic scavenger function is shared between Kupffer cells, which eliminate insoluble waste by phagocytosis, and LEC, which remove soluble colloidal or macromolecular waste (3). Hypothesizing that the scavenger function of LEC in mammals be represented by similar nonmacrophagic, nonphagocytic scavenger endothelia in animals of all vertebrate classes, we set out to screen animal species from the seven major vertebrate classes for the presence of scavenger cells equivalent of mammalian LEC.The findings from these studies strongly suggest the presence of a nonmacrophagic system of scavenger endothelial cells that clear the blood from an array of circulating waste macromolecules and colloids. These scavenger endothelial cells make up the backbone of the reticuloendothelial system (RES), and thus represent an import...

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Section: Resultsmentioning

confidence: 99%

“…20; B. Hansen, B. Arteta, and B.S., unpublished work). Isolation and culture of cod heart scavenger endothelial cells were accomplished by combined collagenase and trypsin treatment followed by differential sedimentation and selective adherence (22).…”

Section: Labeling Of Soluble and Particular Probes With Fluorescence mentioning

confidence: 99%

Scavenger endothelial cells of vertebrates: A nonperipheral leukocyte system for high-capacity elimination of waste macromolecules

Seternes¹,

Sørensen²,

Smedsrød³

2002

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

140

131

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“…Isolation of cardiac endothelial cells (EC) was performed as published (56,57) with some modifications. Briefly, hearts from 9-to 12-mo-old carp were aseptically removed and transferred to 20 ml of ice-cold cRPMI supplemented with 5 IU/ml heparin (Leo Pharmaceutical Products).…”

Section: Cell Isolationmentioning

confidence: 99%

Receptor-Mediated and Lectin-Like Activities of Carp (Cyprinus carpio) TNF-α

Forlenza

Magez²,

Scharsack³

et al. 2009

The Journal of Immunology

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Functional characterization of TNF-α in species other than mammalian vertebrates is limited, and TNF-α has been studied in a limited number of fish species, primarily in vitro using recombinant proteins. Studies on TNF-α from different fish species so far pointed to several inconsistencies, in particular with respect to some receptor-mediated activities of fish TNF-α, such as the ability to directly activate phagocytes. In the present study a comprehensive analysis of in vitro as well as in vivo biological activities of two isoforms of carp TNF-α was performed. Our results show that carp TNF-α directly primes carp phagocytes and indirectly promotes typical receptor-mediated activities such as phagocyte activation by acting via endothelial cells. Additionally, for the first time in nonmammalian vertebrate species, the lectin-like activity of fish TNF-α homologs was investigated. Our results show an evolutionary conservation of function of this receptor-independent activity of TNF-α not only in cyprinid fish, but also in perciform and salmonid fish. The role of TNF-α in vivo, during infections of carp with the blood parasite Trypanoplasma borreli, was examined using three fundamentally different but complementary approaches: (1) inhibition of TNF-α expression, (2) overexpression of TNF-α, and (3) inhibition of shedding of membrane-bound TNF-α. Our results show that, also in fish, a tight regulation of TNF-α expression is important, since depletion or excess of TNF-α can make an important difference to survival of infection. Finally, we demonstrate a crucial protective role for membrane-bound TNF-α, which has a yet unexploited function in fish.

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“…Isolation and cultivation of atrial endocardial endothelial cells Functionally intact atrial endocardial endothelial cells (aEEC) from cod were purified according to (Koren et al, 1997). In short, the heart was dissected out and perfused with L-15 medium supplied with heparin (10·i.u.·ml -1 ).…”

Section: Histological Preparationsmentioning

confidence: 99%

“…Functional studies indicate that the cod aEECs express a set of at least four types of functional endocytic receptors for this purpose: (i) the collagen ␣ chain receptor (Koren et al, 1997;Smedsrød et al, 1995), (ii) the hyaluronan receptor (Seternes et al, 2001b;Sørensen et al, 1997a), (iii) the mannose receptor (Sørensen et al, 2001) and (iv) the scavenger receptor (Seternes et al, 2001a;Sørensen et al, 1998).…”

Section: Introductionmentioning

confidence: 99%

Specific endocytosis and degradation of naked DNA in the endocardial cells of cod (Gadus morhuaL.)

Seternes

Tonheim

Løvoll

et al. 2007

Journal of Experimental Biology

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SUMMARY DNA vaccines are administered in the form of plasmid DNA (pDNA) carrying a strong promoter and the gene of interest. In this study we investigated the tissue distribution, cellular uptake and the fate of intravenously (i.v.) and intramuscularly (i.m.) injected pDNA in Atlantic cod (Gadus morhuaL.). The anatomical distribution of pDNA was determined using both morphological and radiotracing methods. Cellular uptake and receptor specificity were studied in cultures of cod atrial endocardial endothelial cells (aEEC) and head kidney leukocytes. The short-term fate of the endocytosed pDNA in vivo and in vitro was investigated by Southern blot. Expression of the pDNA (R70pRomiLuc)-derived gene was investigated in cod tissues and cultures of cod aEEC by means of real-time RT-PCR and luciferase activity assay. 125I-labelled pDNA was rapidly eliminated from the blood by the aEEC of the cod heart atrium and ventricle. Co-injection of trace amounts of 125I-labelled pDNA with excess amounts of non-labelled pDNA or formaldehyde-treated albumin (FSA), a ligand for the cod EEC scavenger receptor, significantly inhibited the accumulation of the radiotracer in the heart. The organ to blood ratio of radioactivity after inhibition of the cod EEC scavenger receptor demonstrated that the radioactivity not taken up by the EEC remained in the blood. Fluorescence microscopy of tissue sections from cod injected with fluorescein-labelled pDNA confirmed intracellular uptake of pDNA by the endocardial cells of the atrium and ventricle. In purified cultures of cod aEEC the fluorescein-labelled pDNA was taken up in structures reminiscent of endosomal/lysosomal vesicles. Uptake of 125I-labelled pDNA in cultures of cod aEEC was specific. Incubation of cultures with 125I-labelled pDNA together with excess amounts of FSA and fucoidan, which are molecules also known to bind to the scavenger receptors,reduced the uptake of the pDNA by at least 70%. Mannan, a ligand for the mannose receptor, did not inhibit the uptake of 125I-labelled pDNA. Despite, low uptake of 125I-fluorescein-pDNA in the kidney of the cod, the uptake of pDNA in cultured cod head kidney leukocytes was significant. Southern blot analysis of cod tissues after injection of pDNA and culture of aEEC given 10 μg pDNA per 106 cells demonstrated the presence of degradation products in tissues and in the cell cultures. Real-time RT–PCR studies showed expression of luciferase mRNA only at the injection site 168 h after injection. Neither expression of luciferase mRNA nor luciferase activity was present in cod aEEC incubated for 48 h with 10μg pDNA. These results suggest that the EEC are very important for removal of blood borne pDNA in cod and that the uptake by these cells was mediated in a scavenger–receptor-like manner. Uptake of pDNA by head kidney leukocytes was only observed in vitro. The endocytosed DNA was subjected to intracellular degradation and was not expressed by the cod EEC. Despite the low amount of radioactivity found in the head kidney after i.v. injection of 125I-labelled pDNA, the head kidney leukocytes seem to have a high capacity for uptake of 125I-labelled pDNA in vitro.

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Isolation and culture of endocardial endothelial cells from Atlantic salmon (

Cited by 24 publications

References 31 publications

Scavenger endothelial cells of vertebrates: A nonperipheral leukocyte system for high-capacity elimination of waste macromolecules

Scavenger endothelial cells of vertebrates: A nonperipheral leukocyte system for high-capacity elimination of waste macromolecules

Receptor-Mediated and Lectin-Like Activities of Carp (Cyprinus carpio) TNF-α

Specific endocytosis and degradation of naked DNA in the endocardial cells of cod (Gadus morhuaL.)

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