Molecular Characterization of the RNA-Binding Protein Quaking-a in Megalobrama amblycephala: Response to High-Carbohydrate Feeding and Glucose/Insulin/Glucagon Treatment.
Molecular Characterization of the RNA-Binding Protein Quaking-a in Megalobrama amblycephala: Response to High-Carbohydrate Feeding and Glucose/Insulin/Glucagon Treatment.
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团头鲂RNA结合蛋白Quak-a的分子特征：对高碳水化合物喂养和葡萄糖/胰岛素/胰高血糖素治疗的反应
DOI：
10.3389/fphys.2018.00434
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发表时间：
2018
影响因子：
4
通讯作者：
Li XF
中科院分区：
文献类型：
--
作者：
Shi HJ;Liu WB;Xu C;Zhang DD;Wang BK;Zhang L;Li XF
采用cDNA末端快速扩增法从团头鲂肝脏中克隆了RNA结合蛋白quaking-a（Qkia），并对其在葡萄糖代谢中的作用进行了研究。qkia全长cDNA全长1，718 bp，开放阅读框为1，572 bp，编码383个AA。序列比对和系统发育分析表明，大多数鱼类和其他高等脊椎动物之间的高度保守性（97-99%）。在所有检查的器官/组织中检测到qkia的mRNA。然后，分别在腹膜内注射葡萄糖[1.67 g/kg体重（BW）]、胰岛素（0.052 mg/kg BW）和胰高血糖素（0.075 mg/kg BW）的鱼中以及在喂食两种饮食碳水化合物水平（31和41%）的鱼中测定血浆葡萄糖水平和组织qkia表达12周。葡萄糖给药诱导血浆葡萄糖显著增加，在1 h记录到最高值。此后，其降低至基础值。葡萄糖给药后，肝脏和肌肉中qkia的表达显著降低，在1h时达到最低值，脑中qkia的表达在8h时达到最低值。然后逐渐恢复到基础值。胰岛素注射诱导血浆葡萄糖显著降低，在1 h时记录到最低值，而胰高血糖素负荷后情况正好相反（在4 h时获得最高值）。随后，葡萄糖水平逐渐恢复到基础值。胰岛素给药后，qkia在脑和肌肉中的表达显著降低，在2 h达到最低值，在肝脏中的表达在1h达到最低值。而胰高血糖素则显著刺激组织中qkia的表达，在6 h时达到最高值。此外，高碳水化合物水平显著增加血浆葡萄糖水平，但下调组织中qkia的转录。这些结果表明团头鲂与其他脊椎动物的基因具有很高的相似性。葡萄糖和胰岛素的管理，以及高碳水化合物喂养，显着下调其在大脑，肌肉和肝脏的transluctin，而胰高血糖素负荷后，情况正好相反。
The RNA-binding protein quaking-a (Qkia) was cloned from the liver of blunt snout bream Megalobrama amblycephala through the rapid amplification of cDNA ends method, with its potential role in glucose metabolism investigated. The full-length cDNA of qkia covered 1,718 bp, with an open reading frame of 1,572 bp, which encodes 383 AA. Sequence alignment and phylogenetic analysis revealed a high degree of conservation (97–99%) among most fish and other higher vertebrates. The mRNA of qkia was detected in all examined organs/tissues. Then, the plasma glucose levels and tissue qkia expressions were determined in fish intraperitoneally injected with glucose [1.67 g per kg body weight (BW)], insulin (0.052 mg/kg BW), and glucagon (0.075 mg/kg BW) respectively, as well as in fish fed two dietary carbohydrate levels (31 and 41%) for 12 weeks. Glucose administration induced a remarkable increase of plasma glucose with the highest value being recorded at 1 h. Thereafter, it reduced to the basal value. After glucose administration, qkia expressions significantly decreased with the lowest value being recorded at 1 h in liver and muscle and 8 h in brain, respectively. Then they gradually returned to the basal value. The insulin injection induced a significant decrease of plasma glucose with the lowest value being recorded at 1 h, whereas the opposite was true after glucagon load (the highest value was gained at 4 h). Subsequently, glucose levels gradually returned to the basal value. After insulin administration, the qkia expressions significantly decreased with the lowest value being attained at 2 h in brain and muscle and 1 h in liver, respectively. However, glucagon significantly stimulated the expressions of qkia in tissues with the highest value being gained at 6 h. Moreover, high dietary carbohydrate levels remarkably increased plasma glucose levels, but down-regulated the transcriptions of qkia in tissues. These results indicated that the gene of blunt snout bream shared a high similarity with that of the other vertebrates. Glucose and insulin administration, as well as high-carbohydrate feeding, remarkably down-regulated its transcriptions in brain, muscle and liver, whereas the opposite was true after the glucagon load.
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