Four experiments were conducted in this study. Experiment 1 was carried out to examine mRNA expressions of nuclear vitamin D receptor (nVDR), membrane vitamin D receptor (mVDR), and type IIb sodium-phosphate cotransporter (NaPi-IIb) in the small intestine of broiler chickens. Experiments 2, 3, and 4 were implemented to evaluate effects of age, non-phytate phosphorus (NPP), and 25-hydroxycholecalciferol (25-OH-D3) on mRNA expressions of nVDR, mVDR, and NaPi-IIb in the duodenum of chickens. Results showed that mRNA expression levels of nVDR and NaPi-IIb were highest in the duodenum of 21-day-old broilers, lower in the jejunum, and lowest in the ileum. By contrast, no differences in mRNA expression levels of mVDR were detected among the duodenum, jejunum, and ileum. Age quadratically affected mRNA expressions of nVDR, mVDR, and NaPi-IIb in the duodenum and 25-hydroxylase in the liver of 7- to 42-day-old broilers, with the highest levels observed at 21 d of age. By contrast, age linearly decreased mRNA expression level of 1α-hydroxylase in kidneys. Dietary NPP levels quadratically affected mRNA expression levels of nVDR and mVDR in the duodenum and 25-hydroxylase in the liver of 21-day-old broilers. The highest mRNA expression levels of nVDR and mVDR and lowest mRNA level of 25-hydroxylase were observed at 0.55% NPP. mRNA expression level of NaPi-IIb linearly declined when dietary NPP levels increased from 0.25 to 0.65%. Addition of 12.5 μg/kg of 25-OH-D3 increased mRNA expression level of 1α-hydroxylase in kidneys and those of nVDR, mVDR, and NaPi-IIb in the duodenum of broilers compared with birds fed the diet without 25-OH-D3. These data indicate that mRNA expressions of nVDR and NaPi-IIb are highest in the duodenum, and the greatest mRNA levels of nVDR, mVDR, and NaPi-IIb are observed at 21 d of age. Dietary NPP levels quadratically increase mRNA expressions of nVDR and mVDR but linearly decrease NaPi-IIb mRNA level. 25-OH-D3 up-regulates the above gene transcription.