The present study evaluated the mechanisms governing insulin signaling, glucose metabolism, and lipogenesis in juvenile fish fed with different dietary leucine levels. Fish were fed six practical diets with graded leucine levels ranging from 0.90 to 2.94% of dry basis for 8 weeks. The trial results showed that, compared to the control group (0.90%), optimal dietary leucine level (1.72%) resulted in the up-regulation of mRNA expression related to insulin signaling pathway, including target of rapamycin (TOR), insulin receptor substrate 1 (IRS-1), phosphoinositide 3-kinase (PI3K), and protein kinase B (Akt). However, an excessive leucine level (2.94%) led to protein S6 kinase 1 (S6K1) overexpression and inhibited TOR, IRS-1, PI3K, and Akt mRNA expressions. The protein level of TOR, S6K1, IRS-1, PI3K, and Akt showed a similar result with mRNA level of these genes. Optimal dietary leucine level (1.72%) significantly improved plasma insulin content, while high level of leucine showed an inhibiting phenomenon. Optimal dietary leucine level (1.72%) could reduce plasma glucose by enhancing the ability of glycometabolism including improving glucose transporter 2 (GLUT2), glucokinase (GK) expressions and down-regulating phosphoenolpyruvate carboxykinase (PEPCK) expression. While an excessive leucine level (2.94%) resulted in high plasma glucose by inhibiting the ability of glycometabolism including lowering GLUT2 and GK expressions, and improving glucose-6-phosphatase (G6Pase) and PEPCK expressions. The relative expressions of pyruvate kinase (PK) and glycogen synthase (GS) were not significantly affected by dietary leucine levels. Dietary leucine level of 1.33% could improve plasma triglyceride content (TG) by enhancing lipogenesis including improving sterol-response element-binding protein 1 (SREBP1), fatty acid synthase (FAS), acetyl CoA carboxylase (ACC), and glucose-6-phosphate dehydrogenase (G6PDH) expressions compared to the control group (0.90%). Total cholesterol (TC) was not significantly affected by dietary leucine levels. The present results indicate that optimal leucine level could improve glycolysis and fatty acid synthesis through improving insulin sensitivity in juvenile blunt snout bream. However, excessive dietary leucine level resulted in high plasma glucose, which led to insulin resistance by inhibiting the gene expressions of insulin signaling pathway and activating gluconeogenesis-related gene expression.