QUICK SEARCH:   [advanced] Author: Keyword(s):
Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents

This Article Summary Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Seternes, T. Articles by Smedsrød, B. Search for Related Content PubMed PubMed Citation Articles by Seternes, T. Articles by Smedsrød, B.

Scavenger-receptor-mediated endocytosis of lipopolysaccharide in Atlantic cod (Gadus morhua L.)

Tore Seternes^1,*, Roy A. Dalmo², James Hoffman², Jarl Bøgwald², Svetlana Zykova¹ and Bård Smedsrød¹

¹ Department of Experimental Pathology, Institute of Medical Biology, University of Tromsø, N-9037 Tromsø, Norway and
² Institute of Marine Biotechnology, The Norwegian College of Fishery Science, University of Tromsø, N-9037 Tromsø, Norway

View larger version (12K): [in a new window]   Fig. 1. In vitro release of tumour necrosis factor alpha (TNF) from peritoneal macrophages in Balbc mice. TNF was measured in the cell culture supernatants after 24 h of stimulation with lipopolysaccharide (LPS) (1 µg ml–1). The control is supernatant from non-stimulated cultures. The results are presented as the mean + S.E.M. of three independent experiments. F-LPS, fluorescein-labelled LPS.

View larger version (27K): [in a new window]   Fig. 2. Time course of the appearance of radioactivity in tissues of the Atlantic cod (Gadus morhua L.) after intravenous administration of trace amounts of 125I-labelled lipopolysaccharide (0.5–1 µg kg–1 body mass). The results are expressed as means + S.E.M. of three fish per time point.

View larger version (74K): [in a new window]   Fig. 3. Immunohistochemistry on (A) a section of atrium, (B) a section of ventricle and (C) a section of anterior kidney 48 h after intravenous administration of unlabelled lipopolysaccharide. Reaction product (arrows) was observed within endocardial endothelial cells (A,B) and probably in macrophages (C). Scale bars, 10 µm.

View larger version (114K): [in a new window]   Fig. 4. Fluorescence micrograph of atrial endocardial endothelial cells cultured on a glass coverslip (pooled cells from three fish) and incubated with fluorescein-labelled lipopolysaccharide for 1 h at 12°C. All endocardial cells in the monolayer cultures accumulated the probe. Note that fluorescence is confined to discrete vesicles probably representing endocytic vesicles. Scale bar, 10 µm.

View larger version (11K): [in a new window]   Fig. 5. Specificity of endocytosis of 125I-labelled lipopolysaccharide in cultured cod atrial endocardial endothelial cells (aEECs). Monolayer cultures of aEECs in 2 cm2 wells were incubated for 2 h at 12°C with trace amounts of labelled ligand (approximately 2x104 cts min–1; 3 ng) alone (Control) or together with excess amounts of unlabelled macromolecules (100 µg ml–1). The results are presented as a percentage of the control value and are means + S.E.M. of three independent experiments. F-LPS, fluorescein-labelled lipopolysaccharide; FSA, formaldehyde-treated serum albumin.

View larger version (17K): [in a new window]   Fig. 6. Kinetics of endocytosis of 125I-labelled lipopolysaccharide ([125I]LPS) in cultured cod atrial endocardial cells (aEECs). Monolayer cultures of aEECs in 2 cm2 wells were incubated with trace amounts of [125I]LPS (approximately 2x104 cts min–1; 3 ng) at 12°C. The results are presented as the cell-associated percentage of the total added radioactivity and are the results from two independent experiments.

View larger version (15K): [in a new window]   Fig. 7. Intracellular degradation of endocytosed 125I-labelled lipopolysaccharide ([125I]LPS). Monolayer cultures of cod atrial endocardial cells in 2 cm2 wells were incubated for 48 h with [125I]LPS (approximately 2x104 cts min–1; 3 ng). Solubilised cultures were analysed by gel filtration on a PD-10 column. The ordinate indicates the percentage of total eluted radioactivity. The results are mean of six wells ± S.E.M. Vt, total volume.