Abstract
This study aimed to characterize the full-length cDNA of glucose-6-phosphate dehydrogenase (G6PD) from Megalobrama amblycephala with its responses to dietary carbohydrate levels characterized. The cDNA obtained covered 2768 bp with an open reading frame of 1572 bp. Sequence alignment and phylogenetic analysis revealed a high degree of conservation (77–97%) among most fish and other higher vertebrates. The highest transcription of G6PD was observed in kidney followed by liver, whereas relatively low abundance was detected in eye. Then, the transcriptions and activities of G6PD as well as lipid contents were determined in the liver, muscle, and the adipose tissue of fish fed two dietary carbohydrate levels (30 and 42%) for 12 weeks. Hepatic transcriptions of fatty acid synthetase (FAS), acetyl-CoA carboxylase α (ACCα), sterol regulatory element-binding protein-1 (SREBP1), and peroxisome proliferator-activated receptor γ (PPARγ) were also measured to corroborate the lipogenesis derived from carbohydrates. The G6PD expressions and activities in both liver and the adipose tissue as well as the lipid contents in whole-body, liver, and the adipose tissue all increased significantly after high-carbohydrate feeding. Hepatic transcriptions of FAS, ACCα, SREBP1, and PPARγ were also up-regulated remarkably by the intake of a high-carbohydrate diet. These results indicated that the G6PD of M. amblycephala shared a high similarity with that of other vertebrates. Its expressions and activities in tissues were both highly inducible by high-carbohydrate feeding, as also held true for the transcriptions of other enzymes and/or transcription factors involved in lipogenesis, evidencing an enhanced lipogenesis by high dietary carbohydrate levels.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
AOAC (1995) Agricultural chemicals, contaminants, drugs. In: Official methods of analysis of AOAC International. AOAC International, Arlington, p 1298
Bautista JM, Garrido-Pertierra A, Soler G (1988) Glucose-6-phosphate dehydrogenase from Dicentrarchus labrax liver: kinetic mechanism and kinetics of NADPH inhibition. Biochim Biophys Acta 967(3):354–363. https://doi.org/10.1016/0304-4165(88)90098-0
Beydemir Ş, Gülçin İ, Hisar O, Küfrevioğlu Öİ, Yanik T (2005) Effect of melatonin on glucose-6-phosphate dehydrogenase from rainbow trout (Oncorhynchus mykiss) erythrocytes in vitro and in vivo. J Appl Anim Res 28(1):65–68. https://doi.org/10.1080/09712119.2005.9706791
Blasco J, Fernández-Borrás J, Marimón I, Requena A (1996) Plasma glucose kinetics and tissue uptake in brown trout in vivo: effect of an intravascular glucose load. J Comp Physiol B 165(7):534–541. https://doi.org/10.1007/BF00387514
Blasco J, Marimón I, Viaplana I, Fernández-Borrás J (2001) Fate of plasma glucose in tissues of brown trout in vivo: effects of fasting and glucose loading. Fish Physiol Biochem 24(3):247–258. https://doi.org/10.1023/A:101408431
Bou M, Todorčević M, Fontanillas R, Capilla E, Gutiérrez J, Navarro I (2014) Adipose tissue and liver metabolic responses to different levels of dietary carbohydrates in gilthead sea bream (Sparus aurata). Comp Biochem Physiol A Mol Integr Physiol 175(1):72–81. https://doi.org/10.1016/j.cbpa.2014.05.014
Brown MS, Ye J, Rawson RB, Goldstein JL (2000) Regulated intramembrane proteolysis: a control mechanism conserved from bacteria to humans. Cell 100(4):391–398. https://doi.org/10.1016/S0092-8674(00)80675-3
Cappellini MD, Fiorelli G (2008) Glucose-6-phosphate dehydrogenase deficiency. Lancet 371(9606):64–74. https://doi.org/10.1016/S0140-6736(08)60073-2
Choudhary C, Kumar C, Gnad F, Nielsen ML, Rehman M, Walther TC, Olsen JV, Mann M (2009) Lysine acetylation targets protein complexes and co-regulates major cellular functions. Science 325(5942):834–840. https://doi.org/10.1126/science.1175371
Ciardiello MA, Camardella L, Carratore V, di Prisco G (1997) Enzymes in Antarctic fish: glucose-6-phosphate dehydrogenase and glutamate dehydrogenase. Comp Biochem Physiol A Physiol 118(4):1031–1036. https://doi.org/10.1016/S0300-9629(97)86791-6
Ciesla J, Fraczyk T, Rode W (2011) Phosphorylation of basic amino acid residues in proteins: important but easily missed. Acta Biochim Pol 58(2):137–147
Çiftçi M, Beydemir Ş, Yılmaz H, Altıkat S (2003) Purification of glucose 6-phosphate dehydrogenase from Buffalo (Bubalus bubalis) erythrocytes and investigation of some kinetic properties. Protein Expr Purif 29(2):304–310. https://doi.org/10.1016/S1046-5928(03)00073-1
Ciftci M, Ciltas A, Erdogan O (2004) Purification and characterization of glucose 6-phosphate dehydrogenase from rainbow trout (Oncorhynchus mykiss) erythrocytes. Vet Med Czech 49(2):327–333. https://doi.org/10.1016/S1046-5928(03)00073-1
Couto A, Enes P, Peres H, Oliva-Teles A (2008) Effect of water temperature and dietary starch on growth and metabolic utilization of diets in gilthead sea bream (Sparus aurata) juveniles. Comp Biochem Physiol A Mol Integr Physiol 151(1):45–50. https://doi.org/10.1016/j.cbpa.2008.05.013
Craig PM, Moon TW (2013) Methionine restriction affects the phenotypic and transcriptional response of rainbow trout (Oncorhynchus mykiss) to carbohydrate-enriched diets. Brit J Nutr 109(3):402–412. https://doi.org/10.1017/S0007114512001663
Deane EE, Woo NYS (2005) Expression studies on glucose-6-phosphate dehydrogenase in sea bream: effects of growth hormone, somatostatin, salinity and temperature. J Exp Zool A 303((8):676–688. https://doi.org/10.1002/jez.a.201
Ekmann KS, Dalsgaard J, Holm J, Campbell PJ, Skov PV (2013) Glycogenesis and de novo lipid synthesis from dietary starch in juvenile gilthead sea bream (Sparus aurata) quantified with stable isotopes. Brit J Nutr 109(12):2135–2146. https://doi.org/10.1017/S000711451200445X
Enes P, Panserat S, Kaushik S, Oliva-teles A (2008) Growth performance and metabolic utilization of diets with native and waxy maize starch by gilthead sea bream (Sparus aurata) juveniles. Aquaculture 274(1):101–108. https://doi.org/10.1016/j.aquaculture.2007.11.009
Enes P, Sanchez-Gurmaches J, Navarro I, Gutiérrez J, Oliva-Teles A (2010) Role of insulin and IGF-I on the regulation of glucose metabolism in European sea bass (Dicentrarchus labrax) fed with different dietary carbohydrate levels. Comp Biochem Physiol A Mol Integr Physiol 157(4):346–353. https://doi.org/10.1016/j.cbpa.2010.08.006
Erdoğan O, Hisar O, Köroğlu G, Çiltaş A (2005) Sublethal ammonia and urea concentrations inhibit rainbow trout (Oncorhynchus mykiss) erythrocyte glucose-6-phosphate dehydrogenase. Comp Biochem Physiol C 141(2):145–150. https://doi.org/10.1016/j.cca.2005.05.013
Folch J, Lees M, Sloane-Stanley GH (1957) A simple method for the isolation and purification of the total lipid from animal tissue. J Biol Chem 226(1):497–509
Garrett RH, Grisham CM (1996) Biochemistry. Harcourt, Brace & Company, San Diego, p 762
Hemre GI, Mommsen TP, Krogdahl A (2002) Carbohydrates in fish nutrition: effects on growth, glucose metabolism and hepatic enzymes. Aquac Nutr 8(3):175–194. https://doi.org/10.1046/j.1365-2095.2002.00200.x
Hu W, Zhi L, Zhuo MQ, Zhu QL, Zheng JL, Chen QL, Gong Y, Liu CX (2013) Purification and characterization of glucose 6-phosphate dehydrogenase (G6PD) from grass carp (Ctenopharyngodon idella) and inhibition effects of several metal ions on G6PD activity in vitro. Fish Physiol Biochem 39(3):637–647. https://doi.org/10.1007/s10695-012-9726-x
Jiang YY, Li XF, Liu WB, Li HY, He JX (2016) Growth performance, body composition and plasma biochemical parameters of blunt snout bream (Megalobrama amblycephala) yearlings fed practical diets differing in carbohydrate levels. J Nanjing Agric Univ 39(2):281–288 (In Chinese with English Abstract). https://doi.org/10.7685/jnau.201506002
Jin J, Médale F, Kamalam BS, Aguirre P, Véron V, Panserat S (2014) Comparison of glucose and lipid metabolic gene expressions between fat and lean lines of rainbow trout after a glucose load. PLoS One 9(8):e105548. https://doi.org/10.1371/journal.pone.0105548
Kamalam BS, Medale F, Kaushik S, Polakof S, Skiba-Cassy S, Panserat S (2012) Regulation of metabolism by dietary carbohydrates in two lines of rainbow trout divergently selected for muscle fat content. J Exp Biol 215(15):2567–2578. https://doi.org/10.1242/jeb.070581
Katsurada A, Iritani N, Fukuda H, Matsumura Y, Noguchi T, Tanaka T (1989) Effects of nutrients and insulin on transcriptional and post-transcriptional regulation of glucose-6-phosphate dehydrogenase synthesis in rat liver. Biochim Biophys Acta 1006(1):104–110. https://doi.org/10.1016/0005-2760(89)90329-9
Kirchner S, Seixas P, Kaushik S, Panserat S (2005) Effects of low protein intake on extra-hepatic gluconeogenic enzyme expression and peripheral glucose phosphorylation in rainbow trout (Oncorhynchus mykiss). Comp Biochem Physiol B Biochem Mol Biol 140(2):333–340. https://doi.org/10.1016/j.cbpc.2004.10.019
Kumar S, Nei M, Dudley J, Tamura K (2008) MEGA: a biologist-centric software for evolutionary analysis of DNA and protein sequences. Brief Bioinform 9(4):299–306. https://doi.org/10.1093/bib/bbn017
Kuzu M, Aslan A, Ahmed I, Comakli V, Demirdag R, Uzun N (2016) Purification of glucose-6-phosphate dehydrogenase and glutathione reductase enzymes from the gill tissue of Lake Van fish and analyzing the effects of some chalcone derivatives on enzyme activities. Fish Physiol Biochem 42:483–491. https://doi.org/10.1007/s10695-015-0153-7
Lesk AM (1995) NAD-binding domains of dehydrogenases. Curr Opin Struct Biol 5(6):775–783. https://doi.org/10.1016/0959-440X(95)80010-7
Li XF, Liu WB, Lu KL, Xu WN, Wang Y (2012) Dietary carbohydrate/lipid ratios affect stress, oxidative status and non-specific immune responses of fingerling blunt snout bream, Megalobrama amblycephala. Fish Shellfish Immun 33(2):316–323. https://doi.org/10.1016/j.fsi.2012.05.007
Li XF, Wang Y, Liu WB, Jiang GZ, Zhu J (2013) Effects of dietary carbohydrate/lipid ratios on growth performance, body composition and glucose metabolism of fingerling blunt snout bream Megalobrama amblycephala. Aquac Nutr 19(5):701–708. https://doi.org/10.1111/anu.12017
Li XF, Lu KL, Liu WB, Jiang GZ, Xu WN (2014) Effects of dietary lipid and carbohydrate and their interaction on growth performance and body composition of juvenile blunt snout bream, Megalobrama amblycephala. Isr J Aquacult Bamidgeh 66(1):931 http://hdl.handle.net/10524/49111
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25(4):402–408. https://doi.org/10.1006/meth.2001.1262
Mason PJ, Stevens DJ, Luzzatto L, Brenner S, Aparicio S (1995) Genomic structure and sequence of the Fugu rubripes glucose-6-phosphate dehydrogenase gene (G6PD). Genomics 26(3):587–591. https://doi.org/10.1016/0888-7543(95)80179-P
Moon TW (2001) Glucose intolerance in teleost fish: fact or fiction? Comp Biochem Physiol B Biochem Mol Biol 129:243–249. https://doi.org/10.1016/S1096-4959(01)00316-5
Panserat S, Kaushik SJ (2010) Regulation of gene expression by nutritional factors in fish. Aquac Res 41(5):751–762. https://doi.org/10.1111/j.1365-2109.2009.02173.x
Panserat S, Plagnes-Juan E, Kaushik S (2001) Nutritional regulation and tissue specificity of gene expression for proteins involved in hepatic glucose metabolism in rainbow trout (Oncorhynchus mykiss). J Exp Biol 204(13):2351–2360
Panserat S, Skiba-Cassy S, Seiliez I, Lansard M, Plagnes-Juan E, Vachot C, Aguirre P, Larroquet L, Chavernac G, Medale F, Corraze G, Kaushik S, Moon TW (2009) Metformin improves postprandial glucose homeostasis in rainbow trout fed dietary carbohydrates: a link with the induction of hepatic lipogenic capacities? Am J Phys Regul Integr Comp Phys 297(2):707–715. https://doi.org/10.1152/ajpregu.00120.2009
Polakof S, Alvarez R, Soengas JL (2010a) Gut glucose metabolism in rainbow trout: implications in glucose homeostasis and glucosensing capacity. Am J Phys Regul Integr Comp Phys 299(1):19–32. https://doi.org/10.1152/ajpregu.00005.2010
Polakof S, Moon TW, Aguirre P, Skiba-Cassy S, Panserat S (2010b) Effects of insulin infusion on glucose homeostasis and glucose metabolism in rainbow trout fed a high-carbohydrate diet. J Exp Biol 213(24):4151–4157. https://doi.org/10.1242/jeb.050807
Polakof S, Panserat S, Soengas JL, Moon TW (2012) Glucose metabolism in fish: a review. J Comp Physiol B 182(8):1015–1045. https://doi.org/10.1007/s00360-012-0658-7
Polevoda B, Sherman F (2003) N-Terminal acetyltransferases and sequence requirements for N-terminal acetylation of eukaryotic proteins. J Mol Biol 325(24):595–622. https://doi.org/10.1016/S0022-2836(02)01269-X
Prathomya P, Prisingkorn W, Jakovlić I, Deng FY, Zhao YH, Wang WM (2017) 1H NMR-based metabolomics approach reveals metabolic alterations in response to dietary imbalances in Megalobrama amblycephala. Metabolomics 13:17. https://doi.org/10.1007/s11306-016-1158-7
Qian Y, Li XF, Zhang DD, Cai DS, Tian HY, Liu WB (2015) Effects of dietary pantothenic acid on growth, intestinal function, anti-oxidative status and fatty acids synthesis of juvenile blunt snout bream Megalobrama amblycephala. PLoS One 10(3):e0119518. https://doi.org/10.1371/journal.pone.0119518
Qiang J, Yang H, He J, Wang H, Zhu ZX, Xu P (2014) Comparative study of the effects of two high-carbohydrate diets on growth and hepatic carbohydrate metabolic enzyme responses in juvenile gift tilapia (Oreochromis niloticus). Turk J Fish Aquat Sci 14(2):515–525. https://doi.org/10.4194/1303-2712-v14_2_23
Rosemeyer MA (1987) The biochemistry of glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase and glutathione reductase. Cell Biochem Funct 5(2):79–95. https://doi.org/10.1002/cbf.290050202
Sanden M, Frøyland L, Hemre GI (2003) Modulation of glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase and malic enzyme activity by glucose and alanine in Atlantic salmon, Salmo salar L. hepatocytes. Aquaculture 221(1–4):469–480. https://doi.org/10.1016/S0044-8486(03)00077-2
Şentürk M, Ceyhun SB, Erdoğan O, Küfrevioğlu Öİ (2009) In vitro and in vivo effects of some pesticides on glucose-6-phosphate dehydrogenase enzyme activity from rainbow trout (Oncorhynchus mykiss) erythrocytes. Pestic Biochem Phys 95(2):95–99. https://doi.org/10.1016/j.pestbp.2009.07.005
Sheridan MA, Mommsen TP (1991) Effects of nutritional state on in vivo lipid and carbohydrate metabolism of coho salmon, Oncorhynchus kisutch. Gen Comp Endocrinol 81(3):473–483. https://doi.org/10.1016/0016-6480(91)90175-6
Slenzka K, Appel R, Rahmann H (1995) Development and altered gravity dependent changes in glucose-6-phosphate dehydrogenase activity in the brain of the cichlid fish Oreochromis mossambicus. Neurochem Int 26(6):579–585. https://doi.org/10.1016/0197-0186(94)00176-U
Topal A, Atamanalp M, Oruç E, Kırıcı M, Kocaman EM (2014) Apoptotic effects and glucose-6-phosphate dehydrogenase responses in liver and gill tissues of rainbow trout treated with chlorpyrifos. Tissue Cell 46(6):490–496. https://doi.org/10.1016/j.tice.2014.09.001
Walczak R, Tontonoz P (2002) PPARadigms and PPARadoxes: expanding roles for PPARγ in the control of lipid metabolism. J Lipid Res 43(2):177–186
Wang XT, Chan TF, Lam VM, Engel PC (2008) What is the role of the second “structural” NADP+-binding site in human glucose 6-phosphate dehydrogenase? Protein Sci 17(8):1403–1411. https://doi.org/10.1110/ps.035352.108
Wang BK, Liu WB, Xu C, Cao XF, Zhong XQ, Shi HJ, Li XF (2017) Dietary carbohydrate levels and lipid sources modulate the growth performance, fatty acid profiles and intermediary metabolism of blunt snout bream Megalobrama amblycephala in an interactive pattern. Aquaculture 481:140–153. https://doi.org/10.1016/j.aquaculture.2017.08.034
Wilson RP (1994) Utilization of dietary carbohydrate by fish. Aquaculture 124(1–4):67–80. https://doi.org/10.1016/0044-8486(94)90363-8
Xu C, Liu WB, Dai YJ, Jiang GZ, Wang BK, Li XF (2017) Long-term administration of benfotiamine benefits the glucose homeostasis of juvenile blunt snout bream Megalobrama amblycephala fed a high-carbohydrate diet. Aquaculture 470:74–83. https://doi.org/10.1016/j.aquaculture.2016.12.025
Xu C, Liu WB, Zhang DD, Cao XF, Shi HJ, Li XF (2018) Interactions between dietary carbohydrate and metformin: implications on energy sensing, insulin signaling pathway, glycolipid metabolism and glucose tolerance in blunt snout bream Megalobrama amblycephala. Aquaculture 483:183–195. https://doi.org/10.1016/j.aquaculture.2017.10.022
Zar JH (1984) Biostatistical analysis. Prentice-Hall Inc., Englewood Cliffs
Zhang J, Wei XL, Chen LP, Chen N, Li YH, Wang WM, Wang HL (2013) Sequence analysis and expression differentiation of chemokine receptor CXCR4b among three populations of Megalobrama amblycephala. Dev Comp Immunol 40(2):195–201. https://doi.org/10.1016/j.dci.2013.01.011
Funding
This research was funded by the Earmarked Fund for China Agriculture Research System (CARS-45-14) and the Fundamental Research Funds for the Central Universities in China (KJQN201708).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Rights and permissions
About this article
Cite this article
Jiang, GZ., Shi, HJ., Xu, C. et al. Glucose-6-phosphate dehydrogenase in blunt snout bream Megalobrama amblycephala: molecular characterization, tissue distribution, and the responsiveness to dietary carbohydrate levels. Fish Physiol Biochem 45, 401–415 (2019). https://doi.org/10.1007/s10695-018-0572-3
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10695-018-0572-3