Calcium and phosphorus homeostasis is crucial for the performance and bone health of laying hens. The calcium and phosphorus transporters play an important role in calcium and phosphorus absorption, reabsorption, and excretion. In the present study, Hy-Line Brown layers were sampled at brooding period (1, 4, 6 wk), growing and developing period (12, 18 wk) and laying period (20, 28, 80 wk) respectively, and the calcium transporters CaBP-D28k and PMCA1b and phosphorus transporters NPt2a and NPt2b were respectively measured in duodenum, jejunum, ileum and kidney. The result showed that serum calcium increased (P < 0.0001) and phosphorus level fluctuated (P = 0.0019), while alkaline phosphatase activity decreased with age (P < 0.0001). The mRNA and protein expressions of CaBP-D28k in small intestine elevated after maturity (P ≤ 0.0001). In contrast, the PMCA1b mRNA showed a trend to increase with age in jejunum (P = 0.0059) and ileum (P = 0.0825) whereas there was a decrease for PMCA1b protein in 12-18 wk (P ≤ 0.0009). The peak of NPt2b mRNA were observed at 28 wk in duodenum (P = 0.0001) and jejunum (P = 0.0622) and 1 wk in ileum (P < 0.0001). The NPt2b protein expression reached the top point at 4 or 6 wk and 20 wk and decreased to the lowest point around 12 wk (P ≤ 0.0850). In kidney, CaBP-D28k mRNA was not influenced by age (P = 0.4999), while PMCA1b highly expressed in 6-12 wk (P = 0.0003). The protein expressions of CaBP-D28k (P = 0.0148) and PMCA1b (P = 0.0003) decreased with age and lowly expressed in 12-18 wk and increased thereafter. In contrast, NPt2a expression increased steadily with age and decreased at 80 wk (P < 0.0001). In conclusion, the expressions of intestinal calcium and phosphorus transporters were changed by age, intestinal CaBP-D28k and renal NPt2a showed a dramatic increase after maturity, which coincide with the increased calcium and phosphorus requirement for egg production.