Abstract
ABSTRACTThe fatty acid translocase (FAT/CD36) plays an important role in trans-membrane uptake of long-chain fatty acids (LCFA) and has been identified and described in many organisms, while little is known about this gene in freshwater fish, and especially in the grass carp (Ctenopharyngodon idellus). In this study, the uptake of LCFA in hepatocytes of C. idellus was firstly determined and then FAT/CD36 gene was cloned; then the gene was sequenced and mRNA levels from different tissues and the effect of dietary oils on its expression were determined. Present data showed that trans-membrane absorption of LCFA was related to the type and concentration of fatty acids (FAs) and it could be inhibited by specific membrane uptake inhibitors. The FAT/CD36 isolated from grass carp contained 1422 bp and the open reading frame was predicted to have 1403 bp (GenBank access number: KU361231.1). FAT/CD36 was highly expressed in the brain and its lowest expression was in the gills. FAT/CD36 expression in white muscle was up-regulated when fish were fed with groundnut oil, whereas it was down-regulated in fish fed fish and linseed oils, respectively. These results indicated n-3 PUFA might inhibit the FAT/CD36 mRNA expression and therefore the oxidation of FAs.Abbreviations: Docosahexaenoic acid: DHA; Fatty acid: FA; Fatty acid translocase: FAT/CD36; Fish oil: FO; eicosapentaenoic acid: EPA; Groundnut oil: GO; Linseed oil: LO; Peroxisome proliferator-activated nuclear receptor: PPARs; Polyunsaturated fatty acid: PUFA; Unsaturated long-chain fatty acids: LCFA; Olive oil: OO
Highlights
Long-chain fatty acids (LCFAs) are nutrients associated with many important functions of the organism, as they (i) are a main fuel for cellular energy production and a reserve energy source, (ii) are part of phospholipids constituting biological membranes, (iii) may covalently be attached to specific proteins to regulate their function, and (iv) act in selected signal transduction events, (v) and (vi) act to modify gene expression. (Jump 2004; reviewed by Glatz and Luiken 2017)
The experimental approaches used for studying the FA uptake in cells and their metabolism were based on the use of a 14C labelled FA and measuring the intracellular levels of radioactivity (Torstensen and Stubhaug 2004; Stubhaug et al 2005; Vegusdal et al 2005; Zhou et al 2010)
QBT was efficiently taken up by grass carp hepatocytes of grass carp, being highest during the first 45 min post LCFA incubation, whereas this uptake occurred through a saturable transport mechanism (Sorrentino et al 1989), which was in line with the results reported in other Teleost species like in rainbow trout (Richards et al 2003), Atlantic salmon (Zhou et al 2010) as well as in different tissues from higher vertebrates [i.e. skeletal muscle (Turcotte et al 1991), mammalian adipocytes (Abumrad et al 1981; Schwieterman et al 1988), cardiac myocytes (Sorrentino et al 1988) and hepatocytes (Stremmel and Berk 1986)]
Summary
Long-chain fatty acids (LCFAs) are nutrients associated with many important functions of the organism, as they (i) are a main fuel for cellular energy production and a reserve energy source, (ii) are part of phospholipids constituting biological membranes, (iii) may covalently be attached to specific proteins to regulate their function, and (iv) act in selected signal transduction events, (v) and (vi) act to modify gene expression. (Jump 2004; reviewed by Glatz and Luiken 2017). It was considered that LCFAs were taken up by cells through passive diffusion mechanisms (Kamp et al 2003; Pownall and Hamilton 2003; Hamilton and Brunaldi 2007), while evidence has shown that LCFA uptake is facilitated by a proteinmediated mechanism (Abumrad et al 1999; Bonen et al 2007; Guo et al 2013) In this sense, the intracellular transport of free FAs in mammals is facilitated by specific low-molecularweight and highly conserved cytoplasmic proteins that bind both long-chain FAs and other hydrophobic ligands. FAT/CD36-mediated FA transport into the skeletal muscle was reported to be critically coupled with muscle fuel selection and FA oxidation (McFarlan et al 2012; Kim and Lim 2016), probably indicating that an increase in ‘College of Life Science and Technology, Southwest University for Nationalities, Chengdu 610041, People’s Republic of China’
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