Molecular Characterization of 3-Hydroxy-3-Methylglutaryl-Coenzyme A Reductase (HMGR) in Grass Carp Ctenopharyngodon idellus and Its Regulation by Oxidized Fish Oil

3-Hydroxy-3-methylglutaryl-Coenzyme A reductase (HMGR) is a rate-limiting enzyme for cholesterol biosynthesis. The aim of this study was to identify HMGR molecular characterization in grass carp (Ctenopharyngodon idellus), and to evaluate regulation of its expression by oxidized fish oil. Specific primers were designed for 3’RACE and 5’RACE to gain HMGR cDNA sequence of grass carp and to clone full-length HMGR cDNA. This cDNA covered 3591 bp, with 2526 bp for the open reading frame (ORF), 77 bp for 5’-untranslated region (UTR) and 988 bp for 3’UTR. ORF encoded a protein consisting of 841 amino acids. The result revealed that HMGR mRNA is predominantly expressed in liver. Multiple alignments and phylogenetic analysis indicated that grass carp HMGR has a high degree of conservation with other fish species. A 10-week feeding trial was conducted to investigate effects of offering diets containing fresh and oxidized fish oil on HMGR expression in liver and intestine of grass carp. The results showed that HMGR expression in fish fed the diet containing oxidized fish oil was significantly higher than fish fed the fresh fish oil after 10, 20, 30 and 70 days of feeding. Serum diamineoxidase (DAO) activity of fish was also increased by oxidized fish oil.

PDF

___

Chen, X., Wang, X., Li, Z., Kong, L., Liu, G., Fu, J., & Wang, A. (2012a). Molecular cloning, tissue expression and protein structure prediction of the porcine 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMGR) gene. Gene, 495 (2), 170-177. http://dx.doi.org/10.1016/j.gene.2011.12.051

Chen, Y.J., Liu, Y.J., Yang, H.J., Yuan, Y., Liu, F.J., Tian, L.X., Liang, G.Y., & Yuan, R.M. (2012b). Effect of dietary oxidized fish oil on growth performance, body composition, antioxidant defence mechanism and liver histology of juvenile largemouth bass Micropterus salmoides. Aquaculture Nutrition, 18(3), 321–331. https://doi.org/10.1111/j.1365-2095.2011.00900.x

Clarke, P.R., & Hardie, D.G. (1990). Regulation of HMG- CoA reductase: identification of the site phosphorylated by the AMP-activated protein kinase in vitro and in intact rat liver. Embo Journal, 9(8), 2439-2446. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC552270/

Deng, C., Mao, S.Y., Xiong, L., Zhang X.Z., Li, W., Gao, X., Liu, Q.P., Chen, Y., & Liu, Y. (2014). Cloning of Full-length cDNA of HMGR from Gobiocypris rarus and Analysis of Its Expression Profiles in Male Exposed to Pentachlorophenol. Environmental Science, 35(8), 3183-3191. https://doi.org/10.13227/j.hjkx.2014.08.049

Deng, J., Bi, B., Kang, B., Kong, L., Wang, Q., & Zhang, X. (2012). Improving the growth performance and cholesterol metabolism of rainbow trout (Oncorhynchus mykiss) fed soyabean meal-based diets using dietary cholesterol supplementation. British Journal of Nutrition, 110(1), 1-11. https://doi.org/10.1017/S0007114512004680

Gornati, R., Papis, E., Rimoldi, S., Chini, V., Terova, G., Prati, M., Saroglia, M., & Bernardini, G. (2005). Molecular markers for animal biotechnology: sea bass (Dicentrarchus labrax, L.) HMG-CoA reductase mRNA. Gene, 344(3), 299-305. http://dx.doi.org/10.1016/j.gene.2004.11.007

Hentosh, P., Yuh, S.H., Elson, C.E., & Peffley, D.M. (2001). Sterol-independent regulation of 3-hydroxy-3-methylglutaryl coenzyme A reductase in tumor cells. Molecular Carcinogenesis, 32(3), 154-166. https://doi.org/10.1002/mc.1074

Huang, C.H., & Huang, S.L. (2004). Effect of dietary vitamin E on growth, tissue lipid peroxidation, and liver glutathione level of juvenile hybrid tilapia, Oreochromis niloticus × O. aureus, fed oxidized oil. Aquaculture, 237(1-4), 381-389. http://dx.doi.org/10.1016/j.aquaculture.2004.04.002

Huang, Y.W., Ye, Y.T., Cai, C.F., Liu, H.C., Chen, K.Q., Zhang, B.T., Xiao, P.Z., Peng, K., & Xu, D.H. (2015). The effect of the gene expression in metabolism of cholesterol synthesis pathway after intestine injuried on Ctenopharygodon idellus. Journal of Nanjing Agricultural University, 38(3), 497-503. https://doi.org/10.7685/j.issn.1000-2030.2015.03.022

Istvan, E.S., & Deisenhofer, J. (2001). Structural mechanism for statin inhibition of HMG-CoA reductase. Science, 292(5519), 1160-1164. https://doi.org/10.1126/science.1059344

Izquierdo, M.S., Obach, A., Arantzamendi, L., Montero, D., Robaina, L., & Rosenlund, G. (2003). Dietary lipid sources for seabream and seabass: growth performance, tissue composition and flesh quality. Aquaculture Nutrition, 9(6), 397-407. https://doi.org/10.1046/j.1365-2095.2003.00270.x

Li, J.Y., Lv, Y., Fu, X.B., Jin, H., Hu, S., Sun, X.S., & Sheng, Z.S. (2000). The significance of changes in diamine oxidase activity in intestinal injury after trauma. Chinese Critical Care Medicine, 12(8), 482-484.

Lu, K.L., Wang, L.N., Zhang, D.D., Liu, W.B., & Xu, W.N. (2017). Berberine attenuates oxidative stress and hepatocytes apoptosis via protecting mitochondria in blunt snout bream Megalobrama amblycephala fed high-fat diets. Fish Physiology & Biochemistry, 4(1), 65-76. http://doi.org/10.1007/s10695-016-0268-5

Lunt, S.Y., & Heiden, M.G.V. (2011). Aerobic Glycolysis: Meeting the Metabolic Requirements of Cell Proliferation. Annual Review of Cell & Developmental Biology, 27(1), 441-464. https://doi.org/10.1146/annurev-cellbio-092910-154237

Norambuena, F., Lewis, M., Hamid, N.K., Hermon, K., Donald, J.A., & Turchini, G.M. (2013). Fish oil replacement in current aquaculture feed: is cholesterol a hidden treasure for fish nutrition? Plos One, 8(12): e81705. https://doi.org/10.1371/journal.pone.0093135

Olender, E.H., & Simon, R.D. (1992). The intracellular targeting and membrane topology of 3-hydroxy-3-methylglutaryl-CoA reductase. Journal of Biological Chemistry, 267(6), 4223-4235.

Pan, Y., Cohen, I., Guillerault, F., Fève, B., Girard, J., & Pripbuus, C. (2002). The extreme C terminus of rat liver carnitine palmitoyltransferase I is not involved in malonyl-CoA sensitivity but in initial protein folding. Journal of Biological Chemistry, 277(49), 47184-47189. https://doi.org/10.1074/jbc.M208055200

Peng, S., Chen, L., Qin, J.G., Hou, J., Yu, N., Long, Z., Li, E., & Ye, J. (2009). Effects of dietary vitamin E supplementation on growth performance, lipid peroxidation and tissue fatty acid composition of black sea bream (Acanthopagrus schlegeli) fed oxidized fish oil. Aquaculture Nutrition, 15(3), 329-337. http://doi.org/10.1111/j.1365-2095.2009.00657.x

Radhakrishnan, A., Ikeda, Y., Kwon, H.J., Brown, M.S., & Goldstein, J.L. (2007). Sterol-regulated transport of SREBPs from endoplasmic reticulum to Golgi: oxysterols block transport by binding to Insig. Proceedings of the National Academy of Sciences of the United States of America, 104(16), 6511-6518.

Ren, Z.L., & Huo, Q.G. (2000). Effects of oxidized oil on animals. Acta Zoonutrimenta Sinica, 35(7), 46-50.

Romsos, D.R., Allee, G.L., & Leveille, G.A. (1971). In vivo cholesterol and fatty acid synthesis in the pig intestine. Experimental Biology & Medicine, 137(2), 570-573. https://doi.org/abs/10.3181/00379727-137-35623

Sheen, S.S. (2000). Dietary cholesterol requirement of juvenile mud crab Scylla serrata. Aquaculture, 189(3- 4), 277-285. https://doi.org/10.1016/S0044-8486(00)00379-3

Svensson, P.A., Englund, M.C.O., Markström, E., Ohlsson, B.G., Jernås, M., Billig, H., Torgerson, J.S., Wiklund, O., Carlsson, L.M.S., & Carlsson, B. (2003). Copper induces the expression of cholesterogenic genes in human macrophages. Atherosclerosis, 169(1), 71-76. http://dx.doi.org/10.1016/S0021-9150(03)00145-X

Yang, S.P., Liu, H.L., Wang, C.G., Yang, P., Sun, C.B., & Chan, S.M. (2014). Effect of oxidized fish oil on growth performance and oxidative stress of Litopenaeus vannamei. Aquaculture Nutrition, 21(1), 121-127. https://doi.org/10.1111/anu.12143

Yuan Y., Chen Y.J., Liu, Y.J., Yang, H.J., Liang, G.Y., & Tian, L.X. (2014). Dietary high level of vitamin premix can eliminate oxidized fish oil–induced oxidative damage and loss of reducing capacity in juvenile largemouth bass (Micropterus salmoides). Aquaculture Nutrition, 20(2), 109-117. https://doi.org/10.1111/anu.12057

Zager, R.A., & Johnson, A. (2001). Renal cortical cholesterol accumulation is an integral component of the systemic stress response. Kidney International, 60(6), 2299-2310. https://doi.org/10.1046/j.1523-1755.2001.00071.x