Dissociation of <i>Per1</i> and <i>Bmal1</i> circadian rhythms in the suprachiasmatic nucleus in parallel with behavioral outputs.

Abstract

The temporal order of physiology and behavior in mammals is primarily regulated by the circadian pacemaker located in the hypothalamic suprachiasmatic nucleus (SCN). Taking advantage of bioluminescence reporters, we monitored the circadian rhythms of the expression of clock genes <i>Per1</i> and <i>Bmal1</i> in the SCN of freely moving mice and found that the rate of phase shifts induced by a single light pulse was different in the two rhythms. The <i>Per1-luc</i> rhythm was phase-delayed instantaneously by the light presented at the subjective evening in parallel with the activity onset of behavioral rhythm, whereas the <i>Bmal1-ELuc</i> rhythm was phase-delayed gradually, similar to the activity offset. The dissociation was confirmed in cultured SCN slices of mice carrying both <i>Per1-luc</i> and <i>Bmal1-ELuc</i> reporters. The two rhythms in a single SCN slice showed significantly different periods in a long-term (3 wk) culture and were internally desynchronized. Regional specificity in the SCN was not detected for the period of <i>Per1-luc</i> and <i>Bmal1-ELuc</i> rhythms. Furthermore, neither is synchronized with circadian intracellular Ca²⁺ rhythms monitored by a calcium indicator, GCaMP6s, or with firing rhythms monitored on a multielectrode array dish, although the coupling between the circadian firing and Ca²⁺ rhythms persisted during culture. These findings indicate that the expressions of two key clock genes, <i>Per1</i> and <i>Bmal1</i>, in the SCN are regulated in such a way that they may adopt different phases and free-running periods relative to each other and are respectively associated with the expression of activity onset and offset.