Nie Qin, Miao Huijun, Miao Shuyan, Zhou Huihui, Zhang Yanjiao, Zhang Wenbing, Mai Kangsen
The Key Laboratory of Aquaculture Nutrition and Feeds, Ministry of Agriculture, The Key Laboratory of Mariculture (Ministry of Education), Ocean University of China, Qingdao, 266003, People's Republic of China.
Fish Physiol Biochem. 2015 Jun;41(3):819-32. doi: 10.1007/s10695-015-0049-6. Epub 2015 Apr 17.
Glucokinase (GK) and fructose-1,6-bisphosphatase (FBPase) play crucial role in glucose metabolism. In the present study, the cDNA encoding GK and FBPase was cloned from the liver of turbot Scophthalmus maximus by rapid amplification of cDNA end technique. Effects of dietary glucose and dextrin on the activities and gene expressions of these two enzymes were also studied. Results showed that the full length of GK cDNA was 2226 bp, consisting of an open reading frame (ORF) of 1434 bp. The full-length cDNA coding FBPase was 1314 bp with a 1014 bp ORF encoding 337 amino acids. Analyses of gene expression of GK and FBPase were conducted in gill, liver, the whole intestine, the whole kidney, heart, the dorsal white muscle and brain. The highest expression of GK was found in liver, followed by muscle. The expression of FBPase was found higher in liver than heart and gill. Both hepatic GK activity and mRNA expression were highly induced in turbot after being fed with dietary carbohydrates (p < 0.05). However, the GK activity and mRNA expression in the group with dietary glucose did not significantly differ from those in the group with dietary dextrin (p > 0.05). Compared with the control group, there were no significant differences in FBPase activity and mRNA expression in the glucose as well as dextrin group (p > 0.05). The increased hepatic GK activity and gene expression indicated that the first step of glycolysis was activated in turbot by dietary carbohydrates.
葡萄糖激酶(GK)和果糖-1,6-二磷酸酶(FBPase)在葡萄糖代谢中起关键作用。在本研究中,通过cDNA末端快速扩增技术从大菱鲆肝脏中克隆了编码GK和FBPase的cDNA。还研究了饲料中葡萄糖和糊精对这两种酶活性和基因表达的影响。结果表明,GK cDNA全长2226 bp,包含一个1434 bp的开放阅读框(ORF)。编码FBPase的全长cDNA为1314 bp,其1014 bp的ORF编码337个氨基酸。对GK和FBPase的基因表达进行了鳃、肝脏、全肠、全肾、心脏、背白肌和脑的分析。发现GK在肝脏中表达最高,其次是肌肉。FBPase在肝脏中的表达高于心脏和鳃。投喂饲料碳水化合物后,大菱鲆肝脏GK活性和mRNA表达均显著上调(p < 0.05)。然而,饲料葡萄糖组的GK活性和mRNA表达与饲料糊精组无显著差异(p >