Advertisement

n-3 PUFA reduction caused by fabp2 deletion interferes with triacylglycerol metabolism and cholesterolhomeostasis in fish

  • Yan Zhao
  • Xiaojuan Cao
  • Lele Fu
  • Jian GaoEmail author
Genomics, transcriptomics, proteomics

Abstract

Fatty acid-binding protein 2 (Fabp2), which is involved in the transport of long-chain fatty acids, is widely studied in mammals. Nevertheless, the role of this protein in teleost fish is mostly unknown. Here, we produced a fabp2−/− zebrafish (KO) animal model. Compared with wild-type zebrafish (WT), KO had a markedly decreased content of intestinal n-3 poly-unsaturated fatty acids (n-3 PUFAs) and increased levels of intestinal, hepatic, and serum triacylglycerols (TAG). The intestinal transcriptome analysis of KO and WT revealed an obviously disrupted TAG metabolism and up-regulated bile secretion in KO. Expression levels of the genes related to fatty acid transport and cholesterol (CL) absorption in the intestine of KO were significantly lower than those of WT, while the expression levels of genes related to intestinal TAG synthesis and hepatic CL synthesis were in the opposite direction. To confirm these findings, we further established fabp2 transgenic zebrafish (TG). Compared with WT, TG had a markedly increased content of intestinal n-3 PUFAs, a significantly decreased level of hepatic TAG, and significantly higher expression of genes related to fatty acid transport and CL absorption in the intestine. In conclusion, this study suggests that teleost fish fabp2 could promote intestinal n-3 PUFA absorption to mediate TAG synthesis and CL homeostasis, by regulating the genes involved in lipid metabolism.

Keywords

fabp2 Zebrafish Gene knockout and overexpressed Triacylglycerol metabolism Cholesterol homeostasis 

Notes

Funding information

This study was supported by the National Natural Science Foundation of China (31672660 and 31872579), State Key Laboratory of Freshwater Ecology and Biotechnology (Y119011F01), and Fundamental Research Funds for the Central Universities (2662018PY034).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest.

Ethical approval

All zebrafish procedures in this study were in accordance with the recommendations in the Guide for the Institutional Animal Ethics Committee and the use of Laboratory Animals of Huazhong Agricultural University.

Supplementary material

253_2020_10366_MOESM1_ESM.pdf (938 kb)
ESM 1 (PDF 938 kb)

References

  1. André M, Ando S, Ballagny C, Durliat M, Poupard G, Briançon C, Babin PJ (2000) Intestinal fatty acid binding protein gene expression reveals the cephalocaudal patterning during zebrafish gut morphogenesis. Int J Dev Biol 44(2):249–252PubMedGoogle Scholar
  2. Atshaves BP, Foxworth WB, Frolov A, Roths JB, Kier AB, Oetama BK, Piedrahita JA, Schroeder F (1998) Cellular differentiation and I-FABP protein expression modulate fatty acid uptake and diffusion. Am J Phys 274:C633–C644.  https://doi.org/10.1152/ajpcell.1998.274.3.C633 CrossRefGoogle Scholar
  3. Besnard P, Niot I, Poirier H, Clément L, Bernard A (2002) New insights into the fatty acid-binding protein (FABP) family in the small intestine. Mol Cell Biochem 239(1–2):139–147.  https://doi.org/10.1023/A:1020505512364 PubMedCrossRefGoogle Scholar
  4. Carmona-Antoñanzas G, Tocher DR, Martinez-Rubio L, Leaver MJ (2014) Conservation of lipid metabolic gene transcriptional regulatory networks in fish and mammals. Gene 534(1):1–9.  https://doi.org/10.1016/j.gene.2013.10.040 PubMedCrossRefGoogle Scholar
  5. Chang NN, Sun CH, Gao L, Zhu D, Xu XF, Zhu XJ, Xiong JW, Xi JJ (2013) Genome editing with RNA-guided Cas9 nuclease in zebrafish embryos. Cell Res 23:465–472.  https://doi.org/10.1038/cr.2013.45 PubMedPubMedCentralCrossRefGoogle Scholar
  6. Chen XW, Jiang S, Shi ZY (2012) Identification and expression analysis of fabp2 gene from common carp Cyprinus carpio. J Fish Biol 80(3):679–691.  https://doi.org/10.1111/j.1095-8649.2011.03203.x PubMedCrossRefGoogle Scholar
  7. Darimont C, Gradoux N, Persohn E, Cumin F, Pover AD (2000) Effects of intestinal fatty acid-binding protein overexpression on fatty acid metabolism in Caco-2 cells. J Lipid Res 41(1):84–92PubMedGoogle Scholar
  8. D'Aquila T, Hung YH, Carreiro A, Buhman KK (2016) Recent discoveries on absorption of dietary fat: presence, synthesis, and metabolism of cytoplasmic lipid droplets within enterocytes. Biochim Biophys Acta Mol Cell Biol Lipids 1861(8):730–747.  https://doi.org/10.1016/j.bbalip.2016.04.012 CrossRefGoogle Scholar
  9. Esteves A, Knoll-Gellida A, Canclini L, Silvarrey MC, Babin PJ (2015) Fatty acid-binding proteins have the potential to channel dietary fatty acid into enterocyte nuclei. J Lipid Res 57(2):219–232.  https://doi.org/10.1194/jlr.M062232 PubMedCrossRefGoogle Scholar
  10. Eyster KM (2007) The membrane and lipids as integral participants in signal transduction: lipid signal transduction for the non-lipid biochemist. Adv Physiol Educ 31(1):5–16.  https://doi.org/10.1152/advan.00088.2006 PubMedCrossRefGoogle Scholar
  11. Friesen JA, Rodwell VW (2004) The 3-hydroxy-3-methylglutaryl coenzyme-a (hmg-coa) reductases. Genome Biol 5(11):248 http://genomebiology.com/2004/5/11/248 PubMedPubMedCentralCrossRefGoogle Scholar
  12. Gajda AM, Zhou YX, Agellon LB, Fried SK, Kodukula S, Fortson W (2013) Direct comparison of mice null for liver or intestinal fatty acid-binding proteins reveals highly divergent phenotypic responses to high fat feeding. J Biol Che 288(42):30330–30344.  https://doi.org/10.1074/jbc.M113.501676 PubMedCrossRefGoogle Scholar
  13. Gao J, Koshio S, Ishikawa M, Yokoyama S, Mamauag REP, Han Y (2012) Effects of dietary oxidized fish oil with vitamin E supplementation on growth performance and reduction of lipid peroxidation in tissues and blood of red sea bream Pagrus major. Aquaculture 356-357:73–79.  https://doi.org/10.1016/j.aquaculture.2012.05.034 CrossRefGoogle Scholar
  14. Hertzel AV, Bernlohr DA (2000) The mammalian fatty acid-binding protein multigene family: molecular and genetic insights into function. Trends Endocrinol Metabol 11:175–180.  https://doi.org/10.1016/S1043-2760(00)00257-5 CrossRefGoogle Scholar
  15. Kerner J, Hoppel C (2000) Fatty acid import into mitochondria. Biochim Biophys Acta Mol Cell Biol Lipids 1486:1–17.  https://doi.org/10.1016/S1388-1981(00)00044-5 CrossRefGoogle Scholar
  16. Kjær MA, Vegusdal A, Gjoen T, Rustan AC, Todorevic M, Ruyter B (2008) Effect of rapeseed oil and dietary n-3 fatty acids on triacylglycerol synthesis and secretion in Atlantic salmon hepatocytes. Biochim Biophys Acta, Mol Cell Biol Lipids 1781(3):112-122.  https://doi.org/10.1016/j.bbalip.2007.12.004 CrossRefGoogle Scholar
  17. Lai YL, Zhou CY, Huang P, Dong ZY, Mo C, Xie LP, Lin HY, Zhou ZT, Deng GH, Liu Y, Chen YY, Huang SH, Wu ZY, Sun XG, Gao L, Lv ZP (2018) Polydatin alleviated alcoholic liver injury in zebrafish larvae through ameliorating lipid metabolism and oxidative stress. J of Pharmacol 138:46–53.  https://doi.org/10.1016/j.jphs.2018.08.007 CrossRefGoogle Scholar
  18. Leaver MJ, Boukouvala E, Antonopoulou E, Diez A, Favre-Krey L, Ezaz MT (2005) Three peroxisome proliferator-activated receptor isotypes from each of two species of marine fish. Endocrinology 146(7):3150–3162.  https://doi.org/10.1210/en.2004-1638 PubMedCrossRefGoogle Scholar
  19. Leng X, Wu XF, Tian J, Li XQ, Guan L, Weng DC (2012) Molecular cloning of fatty acid synthase from grass carp (Ctenopharyngodon idella) and the regulation of its expression by dietary fat level. Aquac Nutr 18(5):551–558.  https://doi.org/10.1111/j.1365-2095.2011.00917.x CrossRefGoogle Scholar
  20. Li AC, Glass CK (2005) PPAR- and LXR-dependent pathways controlling lipid metabolism and the development of atherosclerosis. J Lipid Res 45(12):2161–2173.  https://doi.org/10.1194/jlr.R400010-JLR200 PubMedCrossRefGoogle Scholar
  21. Liang G, Yang J, Horton JD, Hammer RE, Goldstein JL, Brown MS (2002) Diminished hepatic response to fasting/refeeding and liver X receptor agonists in mice with selective deficiency of sterol regulatory element-binding protein-1c. J Biol Chem 277(11):9520–9528.  https://doi.org/10.1074/jbc.M111421200 PubMedCrossRefGoogle Scholar
  22. Li JX, Yang C, Huang LF, Zeng KW, Cao XJ, Gao J (2019) Inefficient ATP synthesis by inhibiting mitochondrial respiration causes lipids decreased in MSTN lacking muscles of loach Misgurnus anguillicaudatus. Funct Integr Genomics 19(6):889–900.  https://doi.org/10.1007/s10142-019-00688-x PubMedCrossRefGoogle Scholar
  23. Liu Y, Wu G, Han L, Zhao K, Qu Y, Xu A (2015) Association of the FABP2 Ala54Thr polymorphism with type 2 diabetes, obesity, and metabolic syndrome: a population-based case-control study and a systematic meta-analysis. Genet Mol Res 14(1):1155–1168.  https://doi.org/10.4238/2015.February.6.19 PubMedCrossRefGoogle Scholar
  24. Montoudis A, Seidman E, Boudreau F, Beaulieu JF, Menard D, Elchebly M (2008) Intestinal fatty acid binding protein regulates mitochondrion beta-oxidation and cholesterol uptake. J Lipid Res 49(5):961–972.  https://doi.org/10.1194/jlr.M700363-JLR200 PubMedCrossRefGoogle Scholar
  25. Moreno-Mateos MA, Vejnar CE, Beaudoin JD, Fernandez JP, Mis EK, Khokha MK (2015) CRISPRscan: designing highly efficient sgRNAs for CRISPR-Cas9 targeting in vivo. Nat Methods 12:982–988.  https://doi.org/10.1038/nmeth.3543 PubMedPubMedCentralCrossRefGoogle Scholar
  26. Niot I, Poirier H, Tran TTT, Besnard P (2009) Intestinal absorption of long-chain fatty acids: evidence and uncertainties. Prog Lipid Res 48(2):110–115.  https://doi.org/10.1016/j.plipres.2009.01.001 CrossRefGoogle Scholar
  27. Passeri MJ, Cinaroglu A, Gao C, Sadler KC (2009) Hepatic steatosis in response to acute alcohol exposure in zebrafish requires sterol regulatory element binding protein activation. Hepatology 49:443–452.  https://doi.org/10.1002/hep.22667 PubMedPubMedCentralCrossRefGoogle Scholar
  28. Pang SC, Wang HP, Li KY, Zhu ZY, Kang JX, Sun YH (2014) Double transgenesis of humanized fat1 and fat2 genes promotes omega-3 polyunsaturated fatty acids synthesis in a zebrafish model. Mar Biotechnol 16:580–593.  https://doi.org/10.1007/s10126-014-9577-9 PubMedCrossRefGoogle Scholar
  29. Pedroso GL, Hammesm TO, Escobar TDC, Fracasso LB, Forgiarini LF, Da Silveira TR (2012) Blood collection for biochemical analysis in adult zebrafish. J Vis Exp 63:e3865.  https://doi.org/10.3791/3865 CrossRefGoogle Scholar
  30. Pierce M, Wang YM, Denovan-Wright EM, Wright JM (2000) Nucleotide sequence of a cDNA clone coding for an intestinal-type fatty acid binding protein and its tissue-specific expression in zebrafish (Danio rerio). Biochim Biophys Acta, Gene Struct Expression 1490(1):175–183.  https://doi.org/10.1016/S0167-4781(99)00229-8 CrossRefGoogle Scholar
  31. Pishva H, Mahboob, SA, Mehdipour P, Eshraghian MR, Mohammadi-Asl J, Hosseini S, Karimi F (2010) Fatty acid-binding protein-2 genotype influences lipid and lipoprotein response to eicosapentaenoic acid supplementation in hypertriglyceridemic subjects. Nutrition, 26(11-12):1117-1121.  https://doi.org/10.1016/j.nut.2009.09.028 PubMedCrossRefGoogle Scholar
  32. Prows DR, Schroeder F (1997) Metallothionein-IIA promoter induction alters rat intestinal fatty acid binding protein expression, fatty acid uptake, and lipid metabolism in transfected L-cells. Arch Biochem Biophys 340(1):135–143.  https://doi.org/10.1006/abbi.1997.9880 PubMedCrossRefGoogle Scholar
  33. Sargent JR, Tocher DR, Bell JG (2002) The lipids. In: Halver JE, Hardy RW (eds) Fish nutrition, 3rd edn. Academic Press, SanDiego, pp 181–257Google Scholar
  34. Sawicki LR, Arias NMB, Scaglia N, Lockhart LJF, Franchini GR, Storch J, Córsico B (2017) Fabp1 knockdown in human enterocytes impairs proliferation and alters lipid metabolism. Biochim Biophys Acta Mol Cell Biol Lipids 1862(12):1587–1594CrossRefGoogle Scholar
  35. Sekiya M, Yahagi N, Matsuzaka T, Najima Y, Nakakuki M, Nagai R (2003) Polyunsaturated fatty acids ameliorate hepatic steatosis in obese mice by SREBP-1 suppression. Hepatology 38(6):1529–1539.  https://doi.org/10.1016/j.hep.2003.09.028 PubMedCrossRefGoogle Scholar
  36. Sharma B, Gupta V, Dahiya D, Kumar H, Vaiphei K, Agnihotri N (2019) Clinical relevance of cholesterol homeostasis genes in colorectal cancer. Biochim Biophys Acta Mol Cell Biol Lipids 1864(10):1314–1327.  https://doi.org/10.1016/j.bbalip.2019.06.008 PubMedCrossRefGoogle Scholar
  37. Sharma MK, Denovan-Wright EM, Degrave A, Thisse C, Thisse B, Wright JM (2004) Sequence, linkage mapping and early developmental expression of the intestinal-type fatty acid-binding protein gene (fabp2) from zebrafish (Danio rerio). Comp Biochem Physiol Part B, Biochem Mol Bio 138(4):391–398.  https://doi.org/10.1016/j.cbpc.2004.05.009 CrossRefGoogle Scholar
  38. Storch J, McDermott L (2008) Structural and functional analysis of fatty acid-binding proteins. J Lipid Res 50:S126–S131.  https://doi.org/10.1194/jlr.R800084-JLR200 PubMedCrossRefGoogle Scholar
  39. Tocher DR (2003) Metabolism and functions of lipids and fatty acids in teleost fish. Rev Fish Sci 11(2):107–184.  https://doi.org/10.1016/j.jglr.2010.02.004 CrossRefGoogle Scholar
  40. Trapnell C, Williams BA, Pertea G, Mortazavi A, Pachter L (2010) Transcript assembly and quantification by RNA Seq reveals unannotated transcripts and isoform switching during cell differentiation. Nat Biotechnol 28(5):511–515.  https://doi.org/10.1038/nbt.1621 PubMedPubMedCentralCrossRefGoogle Scholar
  41. Vassileva G, Huwyler L, Poirier K, Agellon LB, Toth MJ (2000) The intestinal fatty acid binding protein is not essential for dietary fat absorption in mice. FASEB J 14(13):2040–2046.  https://doi.org/10.1096/fj.99-0959com PubMedCrossRefGoogle Scholar
  42. Venold FF, Penn MH, Jim T, Jinni G, Kortner TM, Ashild K (2013) Intestinal fatty acid binding protein (fabp2) in Atlantic salmon (Salmo salar): localization and alteration of expression during development of diet induced enteritis. Comp Biochem Physiol ParA, Mol Integr Physiol 164(1):229–240.  https://doi.org/10.1016/j.cbpa.2012.09.009 CrossRefGoogle Scholar
  43. Vonk FJ, Minich DM, Verkade H (1997) Intestinal absorption of essential fatty acids under physiological and essential fatty acid-deficient conditions. J Lipid Res 38(9):1709–1721PubMedGoogle Scholar
  44. Wang L, Feng Z, Wang X, Wang X, Zhang X (2010) DEGseq: an R package for identifying differentially expressed genes from RNA-seq data. Bioinformatics 26(1):136–138.  https://doi.org/10.1093/bioinformatics/btp612 PubMedCrossRefGoogle Scholar
  45. Westerfield M (2000) The zebrafish book, a guide for the laboratory use of zebrafish (Danio rerio), OR, 4rd edn. University of Oregon, EugeneGoogle Scholar
  46. Wu K, Huang C, Shi X, Chen F, Xu YH, Pan YX, Luo Z, Liu X (2016) Role and mechanism of the AMPK pathway in waterborne Zn exposure influencing the hepatic energy metabolism of Synechogobius hasta. Sci Rep 6(1):38716.  https://doi.org/10.1038/srep38716
  47. Wu N, Sarna LK, Hwang SY, Zhu Q, Karmin O (2013) Activation of 3-hydroxy-3-methylglutaryl coenzyme a (hmg-coa) reductase during high fat diet feeding. Biochim Biophys Acta (BBA) Mol Basis Dis 1832(10):1560–1568.  https://doi.org/10.1016/j.bbadis.2013.04.024 CrossRefGoogle Scholar
  48. Zhang W, Chen R, Yang T, Xu N, Chen J, Gao Y, Stetler RA (2017) Fatty acid transporting proteins: roles in brain development, aging, and stroke. PLEFA 136:35–45.  https://doi.org/10.1016/j.plefa.2017.04.004 CrossRefGoogle Scholar
  49. Zhang XF, Pang SC, Liu CJ, Wang HP, Ye D, Zhu ZY, Sun YH (2019b) A novel dietary source of EPA and DHA: metabolic engineering of an important freshwater species-common carp by fat1-transgenesis. Mar Biotechnol 21:171–185.  https://doi.org/10.1007/s10126-018-9868-7 PubMedCrossRefGoogle Scholar
  50. Zhang Y, Cao XJ, Gao J (2019a) Cloning of fatty acid-binding protein 2 (fabp2) in loach (Misgurnus anguillicaudatus) and its expression in response to dietary oxidized fish oil. Comp Biochem Physiol Part B, Biochem Mol Biol 229:26–33.  https://doi.org/10.1016/j.cbpb.2018.12.006 CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2020

Authors and Affiliations

  1. 1.College of Fisheries, Key Lab of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education/Key Lab of Freshwater Animal Breeding, Ministry of AgricultureHuazhong Agricultural UniversityWuhanChina
  2. 2.Hubei Provincial Engineering Laboratory for Pond AquacultureWuhanChina

Personalised recommendations