异育银鲫GPRC6A对赖氨酸感知和应答的机制研究

如何提高鱼类对蛋白质尤其是非鱼粉蛋白的利用是水产营养学中亟待解决的问题。哺乳动物研究表明，G蛋白耦联受体C家族6组A亚型（GPRC6A）识别氨基酸后，可能通过ERK-mTOR通路来调控机体代谢，但是其在鱼类氨基酸感知和应答过程中的作用机制不明确。因此，本项目将以异育银鲫为研究对象，首先进行赖氨酸灌喂实验，探究GPRC6A对赖氨酸的响应特征；通过长期养殖实验，探究摄食不同赖氨酸水平饲料后，异育银鲫GPRC6A以及ERK-TOR通路的变化情况；使用过表达、siRNA干扰技术以及添加关键蛋白抑制剂等处理，围绕ERK-TOR通路在肌细胞中探究GPRC6A对赖氨酸的感知和应答机制。本项目旨在基于ERK-TOR通路阐明鱼类GPRC6A在赖氨酸感知和应答过程中的作用机制，为探究鱼类对氨基酸的感知和应答提供理论依据，以期为提高鱼类对蛋白质的利用提供支撑。

How to improve utilization of protein especially non-fishmeal protein in fish is a problem demanding prompt solution in aquatic nutrition. Studies in mammal showed that G protein coupled receptor family C, group 6, subtype A (GPRC6A) could sense the amino acids, and then might regulate metabolism via ERK-mTOR signal pathway. But mechanisms of GPRC6A sensing and responding to amino acids in fish are still unclear. Gibel carp is chosen as the research objects, and the oral administration of lysine is performed firstly, and then the characteristics of GPRC6A responding to lysine are analyzed. Secondly, gibel carp are fed diet with different levels of lysine for 8 w, after which time the expression of GPRC6A and ERK-TOR pathway are tested. At last, then we verify the mechanism of GPRC6A sensing and responding to lysine in muscle cells, through overexpressing or siRNA interfering GPRC6A and adding inhibitors of key proteins. This study is aimed to elucidate the mechanism of GPRC6A sensing and responding to lysine in fish based on ERK-TOR pathway, and then tries to support the study of fish amino acid sensing and responses, and then provides basis for the improvement of protein utilization in fish.