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Molecular cloning and characterization of endoplasmic reticulum stress related genes grp78 and atf6α from black seabream (Acanthopagrus schlegelii) and their expressions in response to nutritional regulation

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Abstract

Glucose-regulated protein 78 (grp78) and activating transcription factor 6α (atf6α) are considered vital endoplasmic reticulum (ER) molecular chaperones and ER stress (ERS) sensors, respectively. In the present study, the full cDNA sequences of these two ERS-related genes were first cloned and characterized from black seabream (Acanthopagrus schlegelii). The grp78 cDNA sequence is 2606 base pair (bp) encoding a protein of 654 amino acids (aa). The atf6α cDNA sequence is 2168 base pair (bp) encoding a protein of 645 aa. The predicted aa sequences of A. schlegelii grp78 and atf6α indicated that the proteins contain all the structural features, which were characteristic of the two genes in other species. Tissues transcript abundance analysis revealed that the mRNAs of grp78 and atf6α were expressed in all measured tissues, but the highest expression of these two genes was all recorded in the gill followed by liver/ brain. Moreover, in vivo experiment found that fish intake of a high lipid diet (HLD) can trigger ERS by activating grp78/Grp78 and atf6α/Atf6α. However, it can be alleviated by dietary betaine supplementation, similar results were also obtained by in vitro experiment using primary hepatocytes of A. schlegelii. These findings will be beneficial for us to evaluate the regulator effects of HLD supplemented with betaine on ERS at the molecular level, and thus provide some novel insights into the functions of betaine in marine fish fed with an HLD.

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Data availability

The data that support the findings of this study are available from the corresponding author upon reasonable request.

Code availability

Not applicable.

Abbreviations

atf6α :

Activating transcription factor 6 α

ER:

Endoplasmic reticulum

ERS:

Endoplasmic reticulum stress

grp78 :

Glucose regulated protein 78

HLD:

High lipid diet

ire1α :

Inositol requiring enzyme-1ɑ

OA:

Oil acid

ORF:

Open reading frame

perk :

RNA-dependent protein kinase-like endoplasmic reticulum kinase

UPM:

Universal Primer A Mix

UPR:

Unfolded protein response

UTR:

Untranslated region.

References

  • Adjoumani JJY, Wang K, Zhou M, Liu W, Zhang D (2017) Effect of dietary betaine on growth performance, antioxidant capacity and lipid metabolism in blunt snout bream fed a high-fat diet. Fish Physiol Biochem 43:1733–1745

    Article  PubMed  CAS  Google Scholar 

  • Bai ZY, Zhu ZY, Wang CM, Xia JH, He XP, Yue GH (2012) Cloning and characterization of the calreticulin gene in Asian seabass (Lates calcarifer). Animal 6:887–893

    Article  PubMed  CAS  Google Scholar 

  • Boujard T, Anne G, Denis C, Geneviève C, Dutto G, Gasset E, Kaushik S (2004) Regulation of feed intake, growth, nutrient and energy utilisation in European sea bass (Dicentrarchus labrax) fed high fat diets. Aquaculture 231:529–545

    Article  Google Scholar 

  • Cao XF, Dai YJ, Liu MY, Yuan XY, Wang CC, Huang YY, Liu W, Jiang GZ (2019) High-fat diet induces aberrant hepatic lipid secretion in blunt snout bream by activating endoplasmic reticulum stress-associated IRE1/XBP1 pathway. Biochim Biophys Acta (BBA) - Mol Cell Biol Lipids 1864:213–223

    CAS  Google Scholar 

  • Craig SAS (2004) Betaine in human nutrition American. Am J Clin Nutr 80:539–549

    Article  PubMed  CAS  Google Scholar 

  • Day CR, Kempson SA (2016) Betaine chemistry, roles, and potential use in liver disease. Biochim Biophys Acta (BBA) - Gen Subjects 1860:1098–1106

    Article  CAS  Google Scholar 

  • Deminice R, Da SRP, Lamarre SG (2015) Betaine supplementation prevents fatty liver induced by a high-fat diet: effects on one-carbon metabolism. Amino acids 47:839–846

    Article  PubMed  CAS  Google Scholar 

  • Dong XD, Xue W, Hua J, Hang Y, Sun L, Miao S, Wei W, Wu X, Du X (2018) Effects of dietary betaine in allogynogenetic gibel carp (Carassius auratus gibelio): enhanced growth, reduced lipid deposition and depressed lipogenic gene expression. Aquac Res 49:1967–1972

    Article  CAS  Google Scholar 

  • Du JJ, Shen LY, Tan ZD, Zhang PW, Zhao X, Xu Y, Gan ML, Yang Q, Ma JD, Jiang A, Tang GQ, Jiang YZ, Jin L, Li MZ, Bai L, Li XW, Wang JY, Zhang SH, Zhu L (2018) Betaine supplementation enhances lipid metabolism and improves insulin resistance in mice fed a high-fat diet. Nutrients 10:131

    Article  PubMed  PubMed Central  Google Scholar 

  • Eklund M, Bauer E, Wamatu J, Mosenthin R (2005) Potential nutritional and physiological functions of betaine in livestock. Nutr Res Rev 18:31–48

    Article  PubMed  CAS  Google Scholar 

  • Fan CY, Wang MX, Ge CX, Wang X, Li JM, Kong LD (2014) Betaine supplementation protects against high-fructose-induced renal injury in rats. J Nutr Biochem 25:353–362

    Article  PubMed  CAS  Google Scholar 

  • Ge CX, Yu R, Xu MX, Li PQ, Fan CY, Li JM, Kong LD (2016) Betaine prevented fructose-induced NAFLD by regulating LXRα/PPARα pathway and alleviating ER stress in rats. Eur J Pharmacol 770:154–164

    Article  PubMed  CAS  Google Scholar 

  • Hazari YM, Habib M, Bashir S, ArifHilal NI, SheikhUl H, EhtishamFazili KM (2016) Natural osmolytes alleviate GRP78 and ATF-4 levels: corroboration for potential modulators of unfolded protein response. Life Sci 146:148–153

    Article  PubMed  CAS  Google Scholar 

  • Heidari R, Niknahad H, Sadeghi A, Mohammadi H, Ghanbarinejad V, Ommati MM, Hosseini A, Azarpira N, Khodaei F, Farshad O (2018) Betaine treatment protects liver through regulating mitochondrial function and counteracting oxidative stress in acute and chronic animal models of hepatic injury. Biomed Pharamacother 103:75–86

    Article  CAS  Google Scholar 

  • Hellemans J, Vandesompele J (2011) qPCR data analysis-unlocking the secret to successful results. PCR Troubleshoot Optim: Essential Guide 1:1–13

  • Hetz C (2012) The unfolded protein response: controlling cell fate decisions under ER stress and beyond. Nat Rev Mol Cell Biol 13:89–102

    Article  PubMed  CAS  Google Scholar 

  • Hollien J (2013) Evolution of the unfolded protein response. Biochim Biophys Acta (BBA) - Mol Cell Biol Lipids 1833:2458–2463

    CAS  Google Scholar 

  • Ishikawa T, Taniguchi Y, Okada T, Takeda S, Mori K (2011) Vertebrate unfolded protein response: mammalian signaling pathways are conserved in medaka fish. Cell Struct Funct 36:247–259

    Article  PubMed  CAS  Google Scholar 

  • Jin M, Pan TT, Cheng X, Zhu TT, Sun P, Zhou F, Ding X, Zhou QC (2019a) Effects of supplemental dietary L-carnitine and bile acids on growth performance, antioxidant and immune ability, histopathological changes and inflammatory response in juvenile black seabream (Acanthopagrus schlegelii) fed high-fat diet. Aquaculture 504:199–209

    Article  CAS  Google Scholar 

  • Jin M, Pan TT, Tocher DR, Betancor MB, Óscar M, Shen Y, Zhu T, Zhou Q, Jiao L, Sun P (2019b) Dietary choline supplementation attenuated high-fat diet-induced inflammation through regulation of lipid metabolism and suppression of NF-κB activation in juvenile black seabream (Acanthopagrus schlegelii). J Nutr Sci 8:e38

    Article  PubMed  PubMed Central  Google Scholar 

  • Jin M, Shen Y, Pan T, Zhu T, Li XJ, Xu FM, Betancor MB, Jiao LF, Tocher DR, Zhou QC (2021) Dietary betaine mitigates hepatic steatosis and inflammation induced by a high-fat-diet by Modulating the Sirt1/Srebp-1/Pparɑ pathway in juvenile black seabream (Acanthopagrus schlegelii). Front Immunol 12:694720

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Jung YS, Sun JK, Kwon DY, Ahn CW, Kim YS, Choi DW, Kim YC (2013) Alleviation of alcoholic liver injury by betaine involves an enhancement of antioxidant defense via regulation of sulfur amino acid metabolism. Food Chem Toxicol 62:292–298

    Article  PubMed  CAS  Google Scholar 

  • Kathirvel E, Morgan K, Nandgiri G, Sandoval BC, Morgan TR (2010) Betaine improves nonalcoholic fatty liver and associated hepatic insulin resistance: a potential mechanism for hepatoprotection by betaine. Am J Physiol-Gastrointest Liver Physiol 299:G1068

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Kaufman RJ, Scheuner D, Schröder M, Shen X, Lee K, Liu CY, Arnold SM (2002) The unfolded protein response in nutrient sensing and differentiation. Nat Rev Mol Cell Biol 3:411–421

    Article  PubMed  CAS  Google Scholar 

  • Kristensen CM, Dethlefsen MM, Tndering AS, Lassen SB, Meldgaard JN, Ringholm S, Pilegaard H (2018) PGC-1α in hepatic UPR during high-fat high-fructose diet and exercise training in mice. Physiol Rep 6:e13819

    Article  PubMed  PubMed Central  Google Scholar 

  • Le Y, Li Y, Jin Y, Jia P, Jia KT, Yi MS (2017) Establishment and characterization of a brain cell line from sea perch, Lateolabrax japonicus. Vitro Cell Dev Biol-Animal 53(9):834–840

    Article  CAS  Google Scholar 

  • Li L, Wang P, Zhao C, Qiu L (2018) The anti-stresses capability of GRP78 in Penaeus monodon: evidence from in vitro and in vivo studies. Fish Shellfish Immunol 72:132–142

    Article  PubMed  CAS  Google Scholar 

  • Liao K, Yan J, Li S, Wang TJ, Xu W, Mai KS, Ai QH (2017) Molecular cloning and characterization of unfolded protein response genes from large yellow croaker (larimichthys crocea) and their expression in response to dietary fatty acids. Comp Biochem Physiol Part B: Biochem Mol Biol 203:53–64

    Article  CAS  Google Scholar 

  • Lin SM, Du P, Huber W, Kibbe WA (2008) Model-based variance-stabilizing transformation for Illumina microarray data. Nucleic Acids Res 36:e11

    Article  PubMed  PubMed Central  Google Scholar 

  • Lin W, Popko B (2009) Endoplasmic reticulum stress in disorders of myelinating cells. Nat Neurosci 12:379–385

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2-△△CT method. Methods 25:402–408

    Article  PubMed  CAS  Google Scholar 

  • Mori K (2009) Signalling pathways in the unfolded protein response: development from yeast to mammals. J Biochem 146:743–750

    Article  PubMed  CAS  Google Scholar 

  • Palade G (1975) Intracellular aspects of the process of protein synthesis. Science 189:347–358

    Article  PubMed  CAS  Google Scholar 

  • Rutkowski D, Wu J, Back S, Callaghan MU, Ferris S, Iqbal J, Clark R, Miao H, Hassler J, Fornek J, Katze M, Hussain M (2008) UPR pathways combine to prevent hepatic steatosis caused by ER stress-mediated suppression of transcriptional master regulators. Dev Cell 15(6):829–840

  • Saitou N, Nei M (1987) The neighbour-joining method: a new method for reconstructing phylogenetic trees. Mol Phylogenet Evol 4:406–425

    CAS  Google Scholar 

  • Schröder M (2008) Endoplasmic reticulum stress responses. Cell Mol Life Sci 65:862–894

    Article  PubMed  Google Scholar 

  • Shen Y, Li X, Bao Y, Zhu T, Wu ZX, Yang BQ, Jiao LF, Zhou QC, Jin M (2022) Differential regulatory effects of optimal or excessive dietary lipid levels on growth, lipid metabolism and physiological response in black seabream (Acanthopagrus schlegelii). Aquaculture 560:738532

    Article  CAS  Google Scholar 

  • Song YF, Luo Z, Huang C, Chen QL, Pan YX, Xu YH (2015) Endoplasmic reticulum stress-related genes in yellow catfish Pelteobagrus fulvidraco: molecular characterization, tissue expression and expression responses to dietary copper deficiency and excess. G3: Genes. Genom Genet 5:2091–2104

    CAS  Google Scholar 

  • Song Z, Deaciuc I, Zhou Z, Song M, Chen T, Hill D, McClain CJ (2007) Involvement of AMP-activated protein kinase in beneficial effects of betaine on high-sucrose diet-induced hepatic steatosis. Am J Physiol-Gastrointest Liver Physiol 293:G894–G902

    Article  PubMed  CAS  Google Scholar 

  • Tabas I, Ron D (2011) Integrating the mechanisms of apoptosis induced by endoplasmic reticulum stress. Nat Cell Biol 13:184–190

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Ueland PM (2011) Choline and betaine in health and disease. J Inherit Metab Dis 34:3–15

    Article  PubMed  CAS  Google Scholar 

  • Wang X, Zhang T, Mao H, Mi YC, Zhong B, Wei LL, Liu XC, Hu CY (2016) Grass carp (Ctenopharyngodon idella) ATF6 (activating transcription factor 6) modulates the transcriptional level of GRP78 and GRP94 in CIK cells. Fish Shellfish Immunol 52:65–73

    Article  PubMed  CAS  Google Scholar 

  • Wang Z, Yao T, Pini M, Zhou ZX, Fantuzzi G, Song ZY (2010) Betaine improved adipose tissue function in mice fed a high-fat diet: a mechanism for hepatoprotective effect of betaine in nonalcoholic fatty liver disease. Am J Physiol-Gastrointest Liver Physiol 298:63

    Article  Google Scholar 

  • Xu L, Huang D, Hu Q, Wu J, Wang YZ, Jie F (2015) Betaine alleviates hepatic lipid accumulation via enhancing hepatic lipid export and fatty acid oxidation in rats fed with a high-fat diet. Br J Nutr 113:1835–1843

    Article  PubMed  CAS  Google Scholar 

  • Yamamoto K, Sato T, Matsui T, Sato M, Okada T, Yoshida H, Harada A, Mori K (2007) Transcriptional induction of mammalian ER quality control proteins is mediated by single or combined action of ATF6α and XBP1. Dev Cell 13(3):365–376

  • Yoshida H (2007) ER stress and diseases. FEBS J 274:630–658

    Article  PubMed  CAS  Google Scholar 

  • Zhao G, He F, Wu C, Pan L, Li NZ, Deng JP, Zhu GQ, Ren WK, Peng YY (2018) Betaine in inflammation: mechanistic aspects and applications. Front Immunol 9:1070

    Article  PubMed  PubMed Central  Google Scholar 

  • Zhou F, Xiao JX, Hua Y, Ngandzali BO, Shao QJ (2011) Dietary l-methionine requirement of juvenile black seabream (Sparus macrocephalus) at a constant dietary cystine level. Aquac Nutr 17:469–481

    Article  CAS  Google Scholar 

  • Zhu Y, Fan Q, Mao H, Liu Y, Hu CY (2013) GRP78 from grass carp (Ctenopharyngodon idella) provides cytoplasm protection against thermal and Pb2+ stress. Fish Shellfish Immunol 34:617–622

    Article  PubMed  CAS  Google Scholar 

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Funding

This research was supported by the National Natural Science Foundation of China (32273142, 31802303), Guangdong Province Key Research and Development Project (2021B0202050001), Fundamental Research Funds for the Provincial Universities of Zhejiang (SJLY2021007), the Open Fund of Zhejiang Provincial Top Key Discipline of Aquaculture in Ningbo University and K. C. Wong Magna Fund in Ningbo University. We would like to thank Dr. Xuemei Duan from the core facility platform of the School of Marine Sciences, Ningbo University, for her technical support during the feeding trial.

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Authors and Affiliations

  1. Laboratory of Fish and Shellfish Nutrition, School of Marine Sciences, Ningbo University, Ningbo, 315211, China

    Yuedong Shen, Wenli Zhao, Yangguang Bao, Jiayun Zhu, Lefei Jiao, Xuemei Duan, Tingting Pan, Qicun Zhou & Min Jin

  2. Key Laboratory of Aquacultural Biotechnology Ministry of Education, Ningbo University, Ningbo, 315211, China

    Yuedong Shen, Wenli Zhao, Yangguang Bao, Jiayun Zhu, Lefei Jiao, Xuemei Duan, Tingting Pan, Qicun Zhou & Min Jin

  3. Instituto de Acuicultura Torre de la Sal (IATS), CSIC, Ribera de Cabanes, 12595, Castellón, Spain

    Óscar Monroig

Authors
  1. Yuedong Shen
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  2. Wenli Zhao
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  10. Min Jin
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Contributions

Qicun Zhou and Min Jin: Conceptualization, Methodology, Validation, and Supervision; Yuedong Shen, Wenli Zhao, Yangguang Bao, and Jiayun Zhu: Formal analysis, Software, Validation; Xuemei Duan, Tingting Pan, and Lefei Jiao: Resources; Yuedong Shen: Writing - Original Draft; Óscar Monroig, Qicun Zhou, and Min Jin: Writing- Reviewing and Editing.

Corresponding authors

Correspondence to Qicun Zhou or Min Jin.

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All animals used in this study were conducted in accordance with Animal Research Institute Committee guidelines of Ningbo University, China, and approved by the Committee of Animal Research Institute, Ningbo University, China.

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Shen, Y., Zhao, W., Bao, Y. et al. Molecular cloning and characterization of endoplasmic reticulum stress related genes grp78 and atf6α from black seabream (Acanthopagrus schlegelii) and their expressions in response to nutritional regulation. Fish Physiol Biochem 49, 1115–1128 (2023). https://doi.org/10.1007/s10695-023-01242-0

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