Objective To investigate the relationship of vitamin D with the intestinal development and study the expression of vitamin D receptor (VDR) in the duodenum of C57BL/6 mice at different developmental stages. Methods Quantitative PCR (qPCR), histology using H&E staining and immunofluorescence staining, and Western blotting (WB) were performed to elucidate the expression of VDR in mice intestine at different growth and developmental stages. Results The peak of VDR mRNA expression reached on 21 d. The pathological result showed that VDR mainly distributed in the cytoplasm of epithelial cells and smooth muscle cells in the mouse duodenum. WB result indicated that there was no nuclear translocation of VDR protein in the mouse duodenum. Conclusions This study demonstrates the regularity of expression of VDR in the mouse duodenum during its development, and contributes to understanding the function of VDR in the intestines.