Analyses of the circadian clock genes expression in whole embryos and maternal major tissues of mice

To create an organism, it is vital to assemble enough cells of the various differentiated types with the correct spatial arrangement within the embryo. Circadian clocks development is closely correlated with all cellular differentiation. However, the expression of its emergence during mammalian development are not fully understood. To determine whether embryonic development is influenced by circadian rhythm, it is necessary to observe the ontogeny of the circadian clock gene. We first measured the expression of key circadian genes in whole embryos and maternal major tissues of 25 female mice using RT-PCR and immunohistochemical analysis. Our results indicated that mouse embryos begin to express key circadian genes and have the capacity to express active circadian regulatory cycles during development. But circadian molecular rhythms can’t be built in embryo. At E15, the expression of Bmal1 , Clock and Per1 mRNA in whole embryo were increased, especially Per1 . In the meanwhile, immunohistochemical analysis shows a small number of PER1 positive cells were observed in the bottom of right atrium. From E16 to E17, CLOCK and PER1 positive cells were observed in the airway smooth muscle, the wall of left atrium and skeletal muscle of body wall. It is interesting that CLOCK and PER1 positive cells could not be detected in the liver. By using RT-PCR, we continue to observe the expression of myogenic regulatory factor in embryos and also analyse the relationship of embryo development and maternal rhythms. From E12, the expression of myogenin increased quickly. The expression of Tcap at E15 significantly increased. myogenin may play a direct role in contributing Tcap expression. The expression of MAZ is always the highest than myogenin and Tcap in embryos. MAZ may concern with the development of skeletal muscle. The clock gene is a positive regulator of myogenesis and the development of organ. In contrast to embryonic tissues, circadian variation was present for Bmal1 , Clock and Per1 at maternal tissues. Our results indicate that circadian clock genes seem to function differently in different tissues of embryo and maternal mice. Synchrony does not occur during embryo development despite exposure to maternal rhythms. But development of embryo may be affected by maternal tissues of mice.