Integration of multi-oscillator circadian clock: in vivo and ex vivo imaging of clock gene expression

我们的目的是阐明中央昼夜节律钟-视交叉上核（SCN）中振荡细胞的相互耦合机制。我们开发了在体内监测系统的时钟基因的表达从SCN的未麻醉和不受限制的动物和一个长期的和实时的离体监测系统的细胞内Ca++水平从整个神经元的培养SCN切片。我们证明，所有SCN神经元表现出强大的昼夜节律的Ca++水平，这是拓扑特异性和之间的耦合的起搏器在背侧和腹侧SCN依赖于突触的相互作用。我们进一步证明了时钟基因Cry对于SCN中振荡细胞网络的发育是必不可少的，因为在断奶期前后发生了连贯的节律表达。

We aimed to clarify mutual coupling mechanisms of oscillator cells in the central circadian clock, the suprachiasmatic nucleus (SCN). We developed in vivo monitoring systems for clock gene expression from the SCN of unanesthetized and unrestrained animals and a long-term and real-time ex vivo monitoring systems for intracellular Ca++ levels from the entire neurons of cultured SCN slices. We demonstrated that all SCN neurons exhibit robust circadian rhythms in Ca++ levels which are topologically specific and the coupling between the pacemakers in the dorsal and ventral SCN depends on synaptic interaction. We further demonstrated that clock gene Cry is essential for the development of oscillator cell network in the SCN for the coherent rhythm expression which takes place around weaning period.