, 1993; Vrang et al., 1995; Kalsbeek et al., 1996; Horvath, 1997; Van der Beek et al., 1997; Buijs et al., 1998; Horvath et al., 1998; Gerhold et al., 2001). In addition to tract-tracing strategies to reveal SCN outputs, there have been a number of studies to exploit novel behavioral patterns that have been found to correlate with altered SCN rhythms. For example, hamsters will spontaneously ‘split’ and exhibit two rest–activity cycles each day instead of one when housed in constant light. In a classic study, de la Iglesia et al. (2003) showed that, in ‘split’ hamsters, the right and left SCN oscillate

out of phase with each click here other, with each SCN’s molecular rhythms in phase with only one of the two daily peaks of activity. Likewise, examination of Per1::GFP expression in cultured SCNs from split mice shows antiphase oscillations that STA-9090 can be monitored for several cycles (Ohta et al., 2005). Subsequent work using this

split model revealed that, rather than a simple right–left split, each SCN splits into two compartments that oscillate in antiphase (Tavakoli-Nezhad & Schwartz, 2005; Yan et al., 2005). This four-way split means that the split hamsters’ SCNs exhibit 24 h rhythms of PER1 protein that cycles in antiphase between the left and right sides and between core and shell subregions. Associated with this SCN oscillation is a 12 h rhythm of FOS expression in brain regions that receive SCN efferents (Butler et al., 2012). In the target regions examined (medial preoptic area, paraventricular

nucleus Branched chain aminotransferase of the hypothalamus, dorsomedial hypothalamus and orexin-A neurons), the oscillations were in-phase between hemispheres (unlike in the SCN), although with detectable right–left differences in amplitude. Importantly, in all three conditions studied (split and unsplit hamsters in constant light, and control hamsters in LD cycles), the timing of FOS expression in targets occurred at the same time of day and always occurred at a common phase reference point of the SCN oscillation, suggesting that, at a specific internal phase, each SCN signals these targets once daily. In addition to communication via direct projections to neural loci, the SCN also sends multisynaptic connections, via the autonomic nervous system, to targets in the periphery, setting the phase of subordinate oscillatory systems and controlling their activity. By applying transynaptic, retrograde viral tracers, such as a pseudo rabies virus, to various organs and glands, precise multisynaptic connections from the SCN to the periphery have been established. Early studies employing this technique established that corticosterone secretion is controlled by direct projections to the adrenal gland (Buijs et al., 1999), lipid mobilization via projections to adipose tissue (Bamshad et al.