摘要Vitamin D3(VD3),an essential nutrient for animals,has been demonstrated to stimulate the uptake of certain amino acids.However,the role of VD3 in the intestine,the primary site for digestion and ab-sorption of nutrients,remains poorly characterized.Here,the grass carp(Ctenopharyngodon idella)was studied to assess the influence of different doses of VD3(15.2,364.3,782.5,1,167.9,1,573.8,and 1,980.1 IU/kg)on growth performance,intestinal morphology,digestive absorption,amino acid transport,and potential signaling molecule levels in a feeding experiment.As a result,dietary VD3 improved growth performance,intestinal structure,and digestive and brush border enzyme activities.Additionally,most intestinal free amino acids and their transporters were upregulated after VD3 intake,except for Ala,Lys,Asp,Leu,solute carrier(SLC)7A7,SLC1A5,and SLC1A3 mRNA in different segments,Leu and SLC6A14 mRNA in the proximal intestine,and SLC7A5 mRNA in the mid and distal intestine.In the crucial target of rapamycin(TOR)signal pathway of amino acid transport,the gene and protein expression of TOR,S6 kinase 1,and activating transcription factor 4 were elevated,whereas 4E-binding protein 1 was decreased,further suggesting an advanced amino acid absorption capacity in the fish due to VD3 sup-plementation.Based on percentage weight gain,feed efficiency,and trypsin activity,the VD3 re-quirements of on-growing grass carp were estimated to be 968.33,1,005.00,and 1,166.67 IU/kg,respectively.Our findings provide novel recommendations for VD3 supplementation to promote diges-tion and absorption capacities of fish,contributing to the overall productivity of aquaculture.