Nihon University College of Bioresource Sciences , Kameino 1866, Fujisawa 252-0880 , Japan.
Biol Open. 2012 Oct 15;1(10):1035-42. doi: 10.1242/bio.20121263. Epub 2012 Aug 21.
Growth hormone (GH) transgenic Amago (Oncorhynchus masou ishikawae), containing the sockeye GH1 gene fused with metallothionein-B promoter from the same species, were generated and the physiological condition through lipid metabolism compared among homozygous (Tg/Tg) and heterozygous GH transgenic (Tg/+) Amago and the wild type control (+/+). Previously, we have reported that the adipose tissue was generally smaller in GH transgenic fish compared to the control, and that the Δ-6 fatty acyl desaturase gene was down-regulated in the Tg/+ fish. However, fatty acid (FA) compositions have not been measured previously in these fish. In this study we compared the FAs composition and content in the liver using gas chromatography. Eleven kinds of FA were detected. The composition of saturated and monounsaturated fatty acids (SFA and MUFA) such as myristic acid (14:0), palmitoleic acid (16:1n-7), and cis-vaccenic acid (cis-18:1n-7) was significantly (P<0.05) decreased in GH transgenic Amago. On the other hand, the composition of polyunsaturated fatty acids (PUFAs) such as linoleic acid (18:2n-6), arachidonic acid (20:4n-6), and docosapentaenoic acid (22:5n-3) was significantly (P<0.05) increased. Levels of serum glucose and triacylglycerol were significantly (P<0.05) decreased in the GH transgenics compared with +/+ fish. Furthermore, 3'-tag digital gene expression profiling was performed using liver tissues from Tg/Tg and +/+ fish, and showed that Mid1 interacting protein 1 (Mid1ip1), which is an important factor to activate Acetyl-CoA carboxylase (ACC), was down-regulated in Tg/Tg fish, while genes involved in FA catabolism were up-regulated, including long-chain-fatty-acid-CoA ligase 1 (ACSL1) and acyl-coenzyme A oxidase 3 (ACOX3). These data suggest that liver tissue from GH transgenic Amago showed starvation by alteration in glucose and lipid metabolism due to GH overexpression. The decrease of serum glucose suppressed Mid1ip1, and caused a decrease of de novo FA synthesis, resulting in a decrease of SFA and MUFA. This induced expression of ACSL1 and ACOX3 to produce energy through β-oxidation in the GH transgenic Amago.
生长激素 (GH) 转基因 Amago(Oncorhynchus masou ishikawae),含有与同种金属硫蛋白-B 启动子融合的虹鳟 GH1 基因,已被生成,并通过脂质代谢比较了纯合子 (Tg/Tg) 和杂合子 GH 转基因 (Tg/+) Amago 与野生型对照 (+/+) 之间的生理状况。之前,我们报道过 GH 转基因鱼的脂肪组织通常比对照小,并且 Δ-6 脂肪酸去饱和酶基因在 Tg/+ 鱼中下调。然而,之前尚未测量过这些鱼的脂肪酸 (FA) 组成。在这项研究中,我们使用气相色谱法比较了肝脏中的 FA 组成和含量。检测到 11 种 FA。饱和脂肪酸 (SFA) 和单不饱和脂肪酸 (MUFA) 的组成,如肉豆蔻酸 (14:0)、棕榈油酸 (16:1n-7) 和顺式- vaccenic 酸 (cis-18:1n-7) 显著降低 (P<0.05) GH 转基因 Amago。另一方面,多不饱和脂肪酸 (PUFA) 的组成,如亚油酸 (18:2n-6)、花生四烯酸 (20:4n-6) 和二十二碳五烯酸 (22:5n-3) 显著增加 (P<0.05)。与+/+ 鱼相比,GH 转基因鱼的血清葡萄糖和三酰甘油水平显著降低 (P<0.05)。此外,使用 Tg/Tg 和+/+ 鱼的肝组织进行了 3'-tag 数字基因表达谱分析,结果表明,作为激活乙酰辅酶 A 羧化酶 (ACC) 的重要因素的 Mid1 相互作用蛋白 1 (Mid1ip1) 在 Tg/Tg 鱼中下调,而参与 FA 分解代谢的基因上调,包括长链脂肪酸-CoA 连接酶 1 (ACSL1) 和酰基辅酶 A 氧化酶 3 (ACOX3)。这些数据表明,由于 GH 过表达,GH 转基因 Amago 的肝组织表现出因葡萄糖和脂质代谢改变引起的饥饿。血清葡萄糖的减少抑制了 Mid1ip1,并导致从头 FA 合成减少,导致 SFA 和 MUFA 减少。这诱导了 ACSL1 和 ACOX3 的表达,通过 GH 转基因 Amago 中的β-氧化产生能量。