Molecular characterization and identification of facilitative glucose transporter 2 (GLUT2) and its expression and of the related glycometabolism enzymes in response to different starch levels in blunt snout bream (Megalobrama amblycephala)
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Facilitative glucose transporters (GLUT) are transmembrane transporters involved in glucose transport across the plasma membrane. In this study, blunt snout bream GLUT2 gene was cloned, and its expression in various tissues and in liver in response to diets with different carbohydrate levels (17.1; 21.8; 26.4; 32.0; 36.3; and 41.9% of dry matter). Blunt snout bream GLUT2 was also characterized. A full-length cDNA fragment of 2577 bp was cloned, which contains a 5′-untranslated region (UTR) of 73 bp, a 3′-UTR of 992 bp, and an open reading frame of 1512 bp that encodes a polypeptide of 503 amino acids with predicted molecular mass of 55.046 kDa and theoretical isoelectric point was 7.52. The predicted GLUT2 protein has 12 transmembrane domains between amino acid residues at 7–29; 71–93; 106–123; 133–155; 168–190; 195–217; 282–301; 316–338; 345–367; 377–399; 412–434; and 438–460. Besides, the conservative structure domains located at 12–477 amino acids belong to the sugar porter family which is the major facilitator superfamily (MFS) of transporters. Blunt snout bream GLUT2 had the high degree of sequence identity to four GLUT2s from zebrafish, chicken, human, and mouse, with 91, 63, 57, and 54% identity, respectively. Quantitative real-time (qRT) PCR assays revealed that GLUT2 expression was high in the liver, intestine, and kidney; highest in the liver and was regulated by carbohydrate intake. Compared with the control group (17.1%), fed by 3 h with higher starch levels (32.0; 36.3; and 41.9%), increased plasma glucose levels and glycemic level went back to basal by 24 h after treatment. Furthermore, higher dietary starch levels significantly increase GLUT2, glucokinase (GK), and pyruvate kinase (PK) expression and concurrently decrease phosphoenolpyruvate carboxykinase (PEPCK) and glucose-6-phosphatase (G6P) mRNA levels (P < 0.05), and these changes were also back to basal levels after 24 h of any dietary treatment. These results indicate that the blunt snout bream is able to regulate their ability to metabolize glucose by improving GLUT2, GK, and PK expression levels and decreasing PEPCK and G6P expression levels.
KeywordsBlunt snout bream Glucose transporter 2 Glycometabolism Starch
We thank Ahmed Mokrani and Hopeson Chisomo-Kasiya for checking and revising our manuscript.
This study was financially supported by the National Natural Science Foundation of Jiangsu Province (BK20161143), the Natural Science Foundation of China, NSFC (31772820), and the Modern Agriculture Industrial Technology System special project—the National Technology System for Conventional Freshwater Fish Industries (CARS-45).
Compliance with ethical standards
The handling of the experimental animal (pre-adult blunt snout bream) is based on the Ministry of Agriculture, China, and international animal welfare laws, guidelines, and policies (FAO 2004).
- Brauge C, Corraze G, Me’dale F (1995) Effect of dietary levels of lipid and carbohydrate on growth performance, body composition, nitrogen excretion and plasma glucose levels in rainbow trout reared at 8 or 18°C. Reprod Nutr Dev 35(3):277–290. https://doi.org/10.1051/rnd:19950304 CrossRefPubMedGoogle Scholar
- Cournarie F, Azzout-Marniche D, Foretz M, Guichard C, Ferre P, Foufelle F (1999) The inhibitory effect of glucose on phosphoenolpyruvate carboxykinase gene expression in cultured hepatocytes is transcriptional and requires glucose metabolism. FEBS Lett 460(3):527–532. https://doi.org/10.1016/S0014-5793(99)01407-6 CrossRefPubMedGoogle Scholar
- Dias J, Rueda-Jasso R, Panserat S, da Conceição LEC, Gomes EF, Dinis MT (2004) Effect of dietary carbohydrate-to-lipid ratios on growth, lipid deposition and metabolic hepatic enzymes in juvenile Senegalese sole (Solea senegalensis, Kaup). Aquac Res 35(12):1122–1130. https://doi.org/10.1111/j.1365-2109.2004.01135.x CrossRefGoogle Scholar
- Enes P, Panserat S, Kaushik S, Oliva-Teles A (2006) Effect of normal and waxy maize starch on growth, food utilization and hepatic glucose metabolism in European sea bass (Dicentrarchus labrax) juveniles. Comp Biochem Physiol A 143(1):89–96. https://doi.org/10.1016/j.cbpa.2005.10.027 CrossRefGoogle Scholar
- FAO 2004 In: Spreij M (ed) National Aquaculture Legislation Overview—China. National Aquaculture Legislation Overview (NALO) Fact Sheets. FAO Fisheries and Aquaculture Department, Rome, Italy (http://www.fao.org/fishery/legalframework/nalo_china/en#tcNB0041)
- Habte-Tsion HM, Ge XP, Liu B, Xie J, Ren MC, Zhou QL, Miao LH, Pan LK, Chen RL (2015) A deficiency or an excess of dietary threonine level affects weight gain, enzyme activity, immune response and immune-related gene expression in juvenile blunt snout bream (Megalobrama amblycephala). Fish Shellfish Immunol 42(2):439–446. https://doi.org/10.1016/j.fsi.2014.11.021 CrossRefPubMedGoogle Scholar
- Hall JR, Short CE, Driedzic WR (2006) Sequence of Atlantic cod (Gadus morhua) GLUT4, GLUT2 and GPDH: developmental stage expression, tissue expression and relationship to starvation-induced changes in blood glucose. J Exp Biol 209(22):4490–4502. https://doi.org/10.1242/jeb.02532 CrossRefPubMedGoogle Scholar
- Joost HG, Bell GI, Best JD, Birnbaum MJ, Charron MJ, Chen YT, Doege H, James DE, Lodish H, Moley KH, Moley JF, Mueckler M, Rogers S, Schurmann A, Seino S, Thorens B (2002) Nomenclature of the GLUT/SLC2A family of sugar/polyol transport facilitators. Am J Physiol Endocrinol Metab 282(4):E974–E976. https://doi.org/10.1152/ajpendo.00407.2001 CrossRefPubMedGoogle Scholar
- Li XF, Liu WB, Jiang YY, Zhu H, Ge XP (2010) Effects of dietary protein and lipid levels in practical diets on growth performance and body composition of blunt snout bream (Megalobrama amblycephala) fingerlings. Aquaculture 303(1):65–70. https://doi.org/10.1016/j.aquaculture.2010.03.014 CrossRefGoogle Scholar
- Liang HL, Ren MC, Habte-Tsion HM, Ge XP, Xie J, Mi HF, Xi BW, Miao LH, Liu B, Zhou QL, Fang W (2016) Dietary arginine affects growth performance, plasma amino acid contents and gene expressions of the TOR signaling pathway in juvenile blunt snout bream, Megalobrama amblycephala. Aquaculture 461:1–8. https://doi.org/10.1016/j.aquaculture.2016.04.009 CrossRefGoogle Scholar
- Liu HL, Wang JT, Wan WJ, Fu PS, Sun MM, Wang HW (2014) Expression of glucose transporter 4 and glucose transporter 2 in different tissues of tilapia and its response to glucose injection. Chin J Anim Nutr 26(11):3500–3509Google Scholar
- Metón I, Mediavilla D, Caseras A, Cantó E, Fernández F, Baanante IV (1999) Effect of diet composition and ration size on key enzyme activities of glycolysis–gluconeogenesis, the pentose phosphate pathway and amino acid metabolism in liver of gilthead sea bream (Sparus aurata). Br J Nutr 82(3):223–232PubMedGoogle Scholar
- Palmer TN, Ryman BE (1972) Studies on oral glucose intolerance in fish. J Fish Biol 4(2):311–319. https://doi.org/10.1111/j.1095-8649.1972.tb05680.x CrossRefGoogle Scholar
- Panserat S, Médale F, Brèque J, Plagnes-Juan E, Kaushik S (2000a) Lack of significant long-term effect of dietary carbohydrate on hepatic glucose-6-phosphatase expression in rainbow trout (Oncorhynchus mykiss). J Nutr Biochem 11(1):22–29. https://doi.org/10.1016/S0955-2863(99)00067-4 CrossRefPubMedGoogle Scholar
- Panserat S, Médale F, Blin C, Brèque J, Vachot C, Plagnes-Juan E, Gomes E, Krishnamoorthy R, Kaushik S (2000b) Hepatic glucokinase is induced by dietary carbohydrate in rainbow trout, gilthead seabream, and common carp. Am J Physiol Regul Integr Comp Physiol 278(5):R1164–R1170. https://doi.org/10.1152/ajpregu.2000.278.5.R1164 CrossRefPubMedGoogle Scholar
- Panserat S, Plagnes-Juan E, Kaushik S (2002) Gluconeogenic enzyme gene expression is decreased by dietary carbohydrate in common carp (Cyprinus carpio) and gilthead seabream (Sparus aurata). Biochim Biophys Acta 1579(1):35–42. https://doi.org/10.1016/S0167-4781(02)00501-8 CrossRefPubMedGoogle Scholar
- Ren MC, Ai QH, Mai KS, Ma HM, Wang XJ (2011) Effect of dietary carbohydrate level on growth performance, body composition, apparent digestibility coefficient and digestive enzyme activities of juvenile cobia, Rachycentron canadum L. Aquac Res 42(10):1467–1475. https://doi.org/10.1111/j.1365-2109.2010.02739.x CrossRefGoogle Scholar
- Ren MC, Habte-Tsion HM, Liu B, Xie J, Ge XP, Zhou QL, Pan LK (2015a) Food deprivation of blunt snout bream, Megalobrama amblycephala fingerlings and the subsequent effect of feeding with different dietary starch levels on glucose metabolism. Isr J Aquacult Bamidgeh 67:9Google Scholar
- Ren MC, Habte-Tsion HM, Xie J, Liu B, Zhou QL, Ge XP, Pan LK, Chen RL (2015b) Effects of dietary carbohydrate source on growth performance, diet digestibility and liver glucose enzyme activity in blunt snout bream, Megalobrama amblycephala. Aquaculture 438:75–81. https://doi.org/10.1016/j.aquaculture.2015.01.008 CrossRefGoogle Scholar
- Sun SM, Gu ZM, Fu HT, Zhu J, Ge XP, Xuan FJ (2016) Molecular cloning, characterization, and expression analysis of p53 from the oriental river prawn, Macrobrachium nipponense, in response to hypoxia. Fish Shellfish Immunol 54:68–76. https://doi.org/10.1016/j.fsi.2016.03.167 CrossRefPubMedGoogle Scholar
- Wood IS, Wang B, Lorente-Cebrián S, Trayhurn P (2007) Hypoxia increases expression of selective facilitative glucose transporters (GLUT) and 2-deoxy-D-glucose uptake in human adipocytes. Biochem Biophys Res Commun 361(2):468–473. https://doi.org/10.1016/j.bbrc.2007.07.032 CrossRefPubMedGoogle Scholar
- Yang Y (2011) Effect of dietary carbohydrate level on growth performance and mRNA expression of several carbohydrate metabolism genes in juvenile Darkbarbel catfish, Pelteobagrus vachelli. East China Normal University, ShanghaiGoogle Scholar
- Zhang Z, Wu RS, Mok HO, Wang Y, Poon WL, Cheng SH, Kong RY (2003) Isolation, characterization and expression analysis of a hypoxia-responsive glucose transporter gene from the grass carp, Ctenopharyngodon idellus. Eur J Biochem 270(14):3010–3017. https://doi.org/10.1046/j.1432-1033.2003.03678.x CrossRefPubMedGoogle Scholar
- Zhou CP, Ge XP, Liu B, Xie J, Chen RL, Ren MC (2015) Effect of high dietary carbohydrate on the growth performance, blood chemistry, hepatic enzyme activities and growth hormone gene expression of wuchang bream (megalobrama amblycephala) at two temperatures. Asian-Australas J Anim Sci 28(2):207–214. https://doi.org/10.5713/ajas.13.0705 CrossRefPubMedPubMedCentralGoogle Scholar