Circadian calcium feeding regime in laying hens related to zinc concentration, gene expression of circadian clock, calcium transporters and oxidative status

The study was conducted to investigate the effects of different circadian calcium feeding regimes on parameters of Zn status and gene expression of circadian clock, calcium transporters and oxidative status in laying hens. In total, 180 of 41-weeks Brown Hy-line laying hens were assigned randomly into three groups, 1-CON group (Control Ca, diets contained 3.4% Ca at both 0730 and 1530 h), 2-HL group (High-low Ca, diets contained 3.6%-3.2% Ca respectively) and 3-LH group (Low-high Ca, diets contained 3.2%-3.6% Ca respectively), which were fed a certain amount of control diet at 0730 h and 1530 h. Blood, tibia, jejunum and kidney samples were collected at 4 h intervals with initial starting at 0800 h after 10 weeks of experiment. Compared with the CON group: 1) the serum zinc in HL group increased at 2000 h, but lower at 1600 h in LH group (P < 0.05). 2) in jejunum, circadian clock genes including CLOCK and BMAL1 expression of HL group were down-regulated at 0000 h and 1600 h, as well as CLOCK, BMAL1, Cry2, Per3 and calcium transporter gene NCX1 in LH group at 2000 h (P < 0.05). 3) in kidney, CLOCK, Cry1, Cry2 and Per3 of LH group were up regulated at 0400 h, CLOCK at 0000 h as well, while CLOCK at 2000 h were down-regulated (P < 0.05). 4) in kidney, the calcium transporters including PMCA, CaBP and CA of LH group were up-regulated at 0000 h, but PMCA and CaBP of LH group were down-regulated at 0800 h, 1200 h and 1600 h, as well as CA at 1600 h and PMCA at 2000 h of LH group (P < 0.05), and the oxidative gene SOD of the LH group was up-regulated at 0400 h, as well as CAT at 0400 h, SOD and GPX1 at 1200 h in HL group, but SOD at 1600 h and 2000 h, and GPX1 at 1600 h were down-regulated in LH group (P < 0.05). These results demonstrated that the dynamic circadian calcium feeding regime affected circadian rhythms of serum zinc concentration as well as the expression of certain genes related to the circadian clock, calcium transport and antioxidative capacity, and circadian calcium feeding regimes may therefore be considered with regard to improving the calcium usability.