Abstract: A formulation composed of vitamin D (V_D), calcium β-hydroxy-β-methylbutyrate (HMBCa), and medium-and-long-chain triacylglycerols (MLCT) in a precise ratio of 1:350000:200000 was examined for its potential impacts on muscle development and enhancement of mobility. A zebrafish larval model of dexamethasone-induced sarcopenia was used to evaluate the formula's influence on muscle tissue morphology, myelin sheath fluorescence intensity, and the expression of muscle atrophy-related genes. Furthermore, a zebrafish fatigue model induced by sodium sulfite was utilized to assess the formula's effects on mobility, as well as levels of Adenosine 5'-triphosphate (ATP), lactate, and reactive oxygen species (ROS) levels, thereby elucidating its impact on zebrafish locomotor behavior and underlying mechanisms. The results indicated that the formulation enhances muscle fiber quality and improves muscle tissue structure in the zebrafish sarcopenia model. Further detailed studies uncovered that the formulation could stimulate myelin regeneration and suppress the expression of muscle atrophy-related genes (mstnb, myod1, trim63a, and fbxo32). Additionally, the concoction increased the activity level of zebrafish in a fatigue model. More detailed investigations revealed that the formulation could boost ATP production, reduce lactate accumulation, and enhance ROS scavenging in zebrafish. In conclusion, the formula demonstrates significant effects in fostering muscle development and enhancing mobility.