Glucose metabolism in fish remains a controversial area of research as many fish species are traditionally considered glucose-intolerant. Although energy homeostasis remodeling has been observed in fish with inhibited fatty acid β-oxidation (FAO), the effects and mechanism of the remodeling caused by blocked glucose uptake remain poorly understood. In this study, we blocked glucose uptake by knocking outglut2in zebrafish. Intriguingly, the complete lethality, found inGlut2-null mice, was not observed inglut2−/−zebrafish. Approxiamately 30% ofglut2−/−fish survived to adulthood and could reproduce. The maternal zygotic mutantglut2(MZglut2) fish exhibited growth retardation, decreased blood and tissue glucose levels, and low locomotion activity. The decreased pancreatic β-cell numbers andinsulinexpression, as well as liverinsulin receptor a(insra), fatty acid synthesis (chrebp, srebf1, fasn, fads2, andscd), triglyceride synthesis (dgat1a), and muscle mechanistic target of rapamycin kinase (mtor) of MZglut2zebrafish, suggest impaired insulin-dependent anabolic metabolism. Upregulated expression of lipolysis (atglandlpl) and FAO genes (cpt1aaandcpt1ab) in the liver and proteolysis genes (bckdk, glud1b, andmurf1a) in muscle were observed in the MZglut2zebrafish, as well as elevated levels of P-AMPK proteins in both the liver and muscle, indicating enhanced catabolic metabolism associated with AMPK signaling. In addition, decreased amino acids and elevated carnitines of the MZglut2zebrafish supported the decreased protein and lipid content of the whole fish. In summary, we found that blocked glucose uptake impaired insulin signaling-mediated anabolismviaβ-cell loss, while AMPK signaling-mediated catabolism was enhanced. These findings reveal the mechanism of energy homeostasis remodeling caused by blocked glucose uptake, which may be a potential strategy for adapting to low glucose levels.