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罗氏沼虾 TOR 的鉴定与特性分析及其在肌肉蛋白和脂质生成中的作用。

Identification and characterization of TOR in Macrobrachium rosenbergii and its role in muscle protein and lipid production.

机构信息

Key Laboratory of Freshwater Aquatic Genetic Resources, Ministry of Agriculture and Rural Affairs, Shanghai Ocean University, Shanghai, 201306, China.

Shanghai Collaborative Innovation for Aquatic Animal Genetics and Breeding, Shanghai, 201306, China.

出版信息

Sci Rep. 2024 Jan 24;14(1):2082. doi: 10.1038/s41598-023-50300-3.

Abstract

The recent scarcity of fishmeal and other resources means that studies on the intrinsic mechanisms of nutrients in the growth and development of aquatic animals at the molecular level have received widespread attention. The target of rapamycin (TOR) pathway has been reported to receive signals from nutrients and environmental stresses, and regulates cellular anabolism and catabolism to achieve precise regulation of cell growth and physiological activities. In this study, we cloned and characterized the full-length cDNA sequence of the TOR gene of Macrobrachium rosenbergii (MrTOR). MrTOR was expressed in all tissues, with higher expression in heart and muscle tissues. In situ hybridization also indicated that MrTOR was expressed in muscle, mainly around the nucleus. RNA interference decreased the expression levels of MrTOR and downstream protein synthesis-related genes (S6K, eIF4E, and eIF4B) (P < 0.05) and the expression and enzyme activity of the lipid synthesis-related enzyme, fatty acid synthase (FAS), and increased enzyme activity of the lipolysis-related enzyme, lipase (LPS). In addition, amino acid injection significantly increased the transcript levels of MrTOR and downstream related genes (S6K, eIF4E, eIF4B, and FAS), as well as triglyceride and total cholesterol tissue levels and FAS activity. Starvation significantly increased transcript levels and enzyme activities of adenylate-activated protein kinase and LPS and decreased transcript levels and enzyme activities of FAS, as well as transcript levels of MrTOR and its downstream genes (P < 0.05), whereas amino acid injection alleviated the starvation-induced decreases in transcript levels of these genes. These results suggested that arginine and leucine activated the TOR signaling pathway, promoted protein and lipid syntheses, and alleviated the pathway changes induced by starvation.

摘要

近年来,鱼粉和其他资源的短缺使得研究水生动物生长和发育过程中营养物质的内在分子机制受到了广泛关注。雷帕霉素靶蛋白(TOR)途径已被报道能接收来自营养物质和环境压力的信号,并调节细胞的合成代谢和分解代谢,以实现对细胞生长和生理活动的精确调控。本研究克隆并鉴定了罗氏沼虾(Macrobrachium rosenbergii)TOR 基因全长 cDNA 序列(MrTOR)。MrTOR 在所有组织中均有表达,在心和肌肉组织中表达量较高。原位杂交结果也表明 MrTOR 在肌肉组织中表达,主要位于细胞核周围。RNA 干扰降低了 MrTOR 及其下游蛋白合成相关基因(S6K、eIF4E 和 eIF4B)的表达水平(P < 0.05),以及脂质合成相关酶脂肪酸合酶(FAS)的表达和酶活性,并增加了脂解相关酶脂肪酶(LPS)的酶活性。此外,氨基酸注射显著增加了 MrTOR 及其下游相关基因(S6K、eIF4E、eIF4B 和 FAS)的转录水平,以及组织中三酰甘油和总胆固醇水平和 FAS 活性。饥饿显著增加了腺苷酸激活蛋白激酶(AMPK)和 LPS 的转录水平和酶活性,降低了 FAS 的转录水平和酶活性,以及 MrTOR 和其下游基因的转录水平(P < 0.05),而氨基酸注射缓解了饥饿引起的这些基因转录水平的降低。这些结果表明精氨酸和亮氨酸激活了 TOR 信号通路,促进了蛋白质和脂质的合成,并缓解了饥饿引起的通路变化。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/73b7/10810085/a46c2ccffbb6/41598_2023_50300_Fig1_HTML.jpg

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