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Characterization of scavenger receptor class B, type I in Atlantic salmon (Salmo salar L.)

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Lipids

Abstract

The scavenger receptor class B, type I (SR-BI) is an important player in regulation of mammalian lipid homeostasis. We therefore wanted to study this receptor in Atlantic salmon (Salmo salar L.), which requires a diet with particular high lipid content. We have for the first time cloned and characterized SR-BI from a salmonid fish. The predicted 494 amino acid protein contained two transmembrane domains, several putative N-glycosylation sites, and showed 72% sequence identity with the predicted homolog from zebrafish. SR-BI expression was analyzed by reverse transcription Real-Time PCR in several tissues, and a high relative expression in salmon midgut was detected, which may suggest that SR-BI has a role in uptake of lipids from the diet. We also expressed a construct of salmon myc-tagged SR-BI in salmon TO cells and HeLa cells, which gave a protein of approximately 80 kDa on reducing SDS-PAGE using an antibody against the myc-epitope. Immunofluorescence microscopy analyses of the salmon SR-BI protein in transiently transfected HeLa cells revealed staining in the cell periphery and in some intracellular membranes, but not in the nucleus, which indicated that the salmon protein may be a functional membrane protein. We also observed a high degree of co-localization using an anti-peptide SR-BI antiserum. We found that 20 μg mL−1 insulin up-regulated the SR-BI mRNA levels in primary cultures of salmon hepatocytes relative to untreated cells. Oleic acid, EPA, DHA, or dexamethasone did not affect the relative expression of SR-BI in this liver model system. In conclusion, the salmon SR-BI cDNA encoded a protein with several features common to those of mammalian species. SR-BI gene expression was high in the intestine, which leads us to propose that SR-BI may contribute to the uptake of lipids from the diet.

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Abbreviations

aa:

amino acid

AB:

antibody

BSA:

bovine serum albumin

CLA-1:

CD36 and LIMPII analogous-1

D:

dermis

Dex:

dexamethasone

DHA:

docosahexaenoic acid

EF1A:

elongation factor 1 alpha

EPA:

eicosapentaenoic acid

FA:

fatty acid

FBS:

fetal bovine serum

G:

gills

H:

heart

HDL:

high-density lipoprotein

HG:

hindgut

HK:

head kidney

Ins:

insulin

L:

liver

LDL:

low-density lipoprotein

MG:

midgut

nt:

nucleotides

OA:

oleic acid

ORF:

open reading frame

PBS:

phosphate buffered saline

PPAR:

peroxisome proliferator-activated receptor

PUFA:

polyunsaturated FA

RACE:

rapid amplification of cDNA ends

REST:

relative expression software tool

S:

spleen

SR-BI/II/III:

scavenger receptor class B, type I/II/III

SDS-PAGE:

sodium dodecyl sulfate polyacrylamide gel electrophoresis

TAG:

triacylglycerol

UTR:

untranslated region

VLDL:

very low-density lipoprotein

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Authors and Affiliations

  1. AKVAFORSK, Institute of Aquaculture Research, Ås, Norway

    B. Ruyter & A. Vegusdal

  2. Department of Pharmaceutical Biosciences, School of Pharmacy, University of Oslo, Blindern, P.O. Box 1068, N-0316, Oslo, Norway

    E. J. Kleveland, B. L. Syvertsen, S. M. Jørgensen & T. Gjøen

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  1. E. J. Kleveland
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  2. B. L. Syvertsen
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  3. B. Ruyter
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  4. A. Vegusdal
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  6. T. Gjøen
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Kleveland, E.J., Syvertsen, B.L., Ruyter, B. et al. Characterization of scavenger receptor class B, type I in Atlantic salmon (Salmo salar L.). Lipids 41, 1017–1027 (2006). https://doi.org/10.1007/s11745-006-5052-3

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