Cheng Ronshan, Chang Kay-Min, Wu Jen-Leih
Department of Aquaculture, National Taiwan Ocean University, Keelung, Taiwan.
Mar Biotechnol (NY). 2002 Jun;4(3):218-25. doi: 10.1007/s10126-002-0014-0.
In vertebrates the insulin-like growth factor-I (IGF-I) is well recognized for its important roles in prenatal and postnatal processes. Liver is the major endocrine organ in mammals to produce IGF-I and its modulators, the insulin-like growth factor binding proteins (IGFBPs), which form a binary complex with IGF-I in circulation or in extracellular fluid. In the complex, IGFBP-3 accounts for the majority of IGFBPs that interact with IGF-I in circulation. To date, we know little of fish IGF-I regulation at the gene or protein level. In this preliminary experiment, we isolated tilapia IGFBP-3 complementary DNA sequence by degenerate polymerase chain reaction cloning. This identified clone contained a partial open reading frame of 635 bp and lacked both 5' and 3' end cDNA sequence information. The deduced 211 amino acid residues shared approximately 50% identity with mammalian counterparts. This tilapia IGFBP-3 transcript was expressed in every tissue examined with the highest level found in liver. In a growth hormone time course experiment, both IGF-I and IGFBP-3 message levels increased sharply, and after 24 hours of injection, IGF-I and IGFBP-3 was increased 1.97 and 2.15 fold, respectively. The relative message ratio of IGF-I to IGFBP-3 was 1.58, 1.09, 0.94, 0.91, and 0.92 for 3, 6, 9, 12, and 24 hours, respectively. After 9 hours of treatment, the IGF-I/IGFBBP-3 message ratio maintained a constant value of approximately 0.9 throughout the experiment. This is the first report to demonstrate a fish counterpart of IGFBP-3 at the cDNA level and its message expression ratio with IGF-I under growth hormone induction.
在脊椎动物中,胰岛素样生长因子-I(IGF-I)在产前和产后过程中的重要作用已得到充分认可。肝脏是哺乳动物产生IGF-I及其调节剂胰岛素样生长因子结合蛋白(IGFBPs)的主要内分泌器官,IGFBPs在循环或细胞外液中与IGF-I形成二元复合物。在该复合物中,IGFBP-3占循环中与IGF-I相互作用的IGFBPs的大部分。迄今为止,我们对鱼类IGF-I在基因或蛋白质水平上的调控了解甚少。在这个初步实验中,我们通过简并聚合酶链反应克隆分离出罗非鱼IGFBP-3互补DNA序列。该鉴定的克隆包含一个635 bp的部分开放阅读框,并且缺乏5'和3'末端cDNA序列信息。推导的211个氨基酸残基与哺乳动物对应物具有约50%的同一性。这种罗非鱼IGFBP-3转录本在所检测的每个组织中均有表达,在肝脏中表达水平最高。在生长激素时间进程实验中,IGF-I和IGFBP-3的信使水平均急剧增加,注射24小时后,IGF-I和IGFBP-3分别增加了1.97倍和2.15倍。IGF-I与IGFBP-3的相对信使比率在3、6、9、12和24小时分别为1.58、1.09、0.94、0.91和0.92。处理9小时后,在整个实验过程中IGF-I/IGFBBP-3信使比率维持在约0.9的恒定值。这是首次在cDNA水平证明鱼类IGFBP-3对应物及其在生长激素诱导下与IGF-I的信使表达比率的报告。