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Cloning and Characterization of Lxr and Srebp1, and Their Potential Roles in Regulation of LC-PUFA Biosynthesis in Rabbitfish Siganus canaliculatus

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Lipids

Abstract

Rabbitfish Siganus canaliculatus was the first marine teleost demonstrated to have the ability to biosynthesize C20–22 long-chain polyunsaturated fatty acid (LC-PUFA) from C18 PUFA precursors, which is generally absent or low in marine teleosts. Thus, understanding the molecular basis of LC-PUFA biosynthesis in rabbitfish will contribute to efforts aimed at optimizing LC-PUFA biosynthesis in teleosts, especially marine species. In the present study, the importance of the transcription factors liver X receptor (Lxr) and sterol regulatory element-binding protein 1 (Srebp1) in regulation of LC-PUFA biosynthesis in rabbitfish was investigated. First, full-length cDNA of Lxr and Srebp1 were cloned and characterized. The Lxr mRNA displayed a ubiquitous tissue expression pattern while Srebp1 was highly expressed in eyes, brain and intestine. In rabbitfish primary hepatocytes treated with Lxr agonist T0901317, the expression of Lxr and Srebp1 was activated, accompanied by elevated mRNA levels of Δ4 and Δ6/Δ5 fatty acyl desaturase (Fad), key enzymes of LC-PUFA biosynthesis, as well as peroxisome proliferator-activated receptor γ (PPARγ). In addition, Srebp1 displayed higher expression levels in liver of rabbitfish fed a vegetable oil diet or reared at 10 ppt salinity, which were conditions reported to increase the liver expression of Δ4 and Δ6/Δ5 Fad and LC-PUFA biosynthetic ability, than fish fed a fish oil diet or reared at 32 ppt, respectively. These results suggested that Lxr and Srebp1 are involved in regulation of LC-PUFA biosynthesis probably by promoting the expression of two Fad in rabbitfish liver, which, to our knowledge, is the first report in marine teleosts.

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Abbreviations

ARA:

Arachidonic acid (20:4n-6)

cDNA:

Complementary deoxyribonucleic acid

DHA:

Docosahexaenoic acid (22:6n-3)

DMSO:

Dimethyl sulfoxide

Elovl:

Elongase of very long-chain fatty acid

EFA:

Essential fatty acid

EPA:

Eicosapentaenoic acid (20:5n-3)

EtOH:

Ethanol

Fad:

Fatty acyl desaturase

FBS:

Fetal bovine serum

FO:

Fish oil

LC-PUFA:

Long-chain polyunsaturated fatty acid

Lxr:

Liver X receptor

NAMBS:

Nan Ao Marine Biology Station

PCR:

Polymerase chain reaction

PPAR:

Peroxisome proliferator-activated receptor

PUFA:

Polyunsaturated fatty acids

Srebp:

Sterol regulatory element binding protein

VO:

Vegetable oil

22HC:

22(R)-Hydroxycholesterol

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Acknowledgments

This work was financially supported by the Major International Joint Research Project from the National Natural Science Foundation of China (NSFC) (31110103913), NSFC General Projects (No. 41276179) and Youth Projects (Nos. 31202011, 31202012).

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Authors and Affiliations

  1. Marine Biology Institute and Guangdong Provincial Key Laboratory of Marine Biotechnology, Shantou University, Shantou, Guangdong, China

    Qinghao Zhang, Cuihong You, Fang Liu, Wendi Zhu, Shuqi Wang, Dizhi Xie & Yuanyou Li

  2. Institute of Aquaculture, University of Stirling, Stirling, Scotland, UK

    Óscar Monroig & Douglas R. Tocher

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  1. Qinghao Zhang
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  2. Cuihong You
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  3. Fang Liu
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Zhang, Q., You, C., Liu, F. et al. Cloning and Characterization of Lxr and Srebp1, and Their Potential Roles in Regulation of LC-PUFA Biosynthesis in Rabbitfish Siganus canaliculatus . Lipids 51, 1051–1063 (2016). https://doi.org/10.1007/s11745-016-4176-3

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