SummaryThe change in irradiance at dawn and dusk provides the primary cue for the entrainment of the mammalian circadian pacemaker. Irradiance detection has been ascribed largely to melanopsin-based phototransduction [1–5]. Here we examine the role of ultraviolet-sensitive (UVS) cones in the modulation of circadian behavior, sleep, and suprachiasmatic nucleus (SCN) electrical activity. UV light exposure leads to phase-shifting responses comparable to those of white light. Moreover, UV light exposure induces sleep in wild-type and melanopsin-deficient (Opn4−/−) mice with equal efficacy. Electrical recordings from the SCN of wild-type mice show that UV light elicits irradiance-dependent sustained responses that are similar to those induced by white light, with characteristic fast transient components occurring at the light transitions. These responses are retained in Opn4−/− mice and preserved under saturating photopic conditions. The sensitivity of phase-shifting responses to UV light is unaffected by the loss of rods but is severely attenuated by the additional loss of cones. Our data show that UVS cones play an important role in circadian and sleep regulation in mice.

Circadian rhythms are regulated by the suprachiasmatic nucleus (SCN), a small structure at the base of the hypothalamus. While light effects on the SCN are well established, little is known of behavioral effects. This study elucidates direct modulating action of behavioral activity on the SCN by use of in vivo electrophysiology recordings, assessments of general locomotor behavior, and video-tracking of mice. The results show suppression of SCN neuronal activity by spontaneous behavior, the magnitude being dependent on the intensity, duration and type of behavioral activity. The suppression was moderate (32% of circadian amplitude) for low-intensity behavior and considerable (59%) for locomotor activity. Mild manipulation of the animals had reversed effects on the SCN indicating that different mechanisms are involved in the regulatory effect of spontaneous versus induced activity. The results indicate that exercise at the proper time of the cycle can boost the amplitude of the rhythm of the SCN clock itself. This has potentially beneficial effects for other rhythmic functions that are under the control of the SCN.

Ligand-binding studies revealed the presence of GnRH-binding sites in African catfish ovary. However, our expression profiling studies failed to detect the previously identified catfish GnRH receptor (cfGnRH-R1) mRNA in this tissue. This negative result instigated us to clone an additional catfish GnRH receptor (cfGnRH-R2) cDNA and study its expression in different tissues in conjunction with the expression of the two catfish GnRH (i.e. cfGnRH and cGnRH-II) genes. The highest cfGnRH-R1 and cfGnRH-R2 mRNA levels were detected in pituitary for cfGnRH-R1 and in brain and ovary for cfGnRH-R2. cfGnRH mRNA was coexpressed with cfGnRH-R1 mRNA in pituitary and brain and with cfGnRH-R2 mRNA in brain and ovary. Ubiquitous expression of cGnRH-II mRNA was observed in all tissues tested, with the highest expression in brain, heart, pituitary, ovary, and head-kidney. Binding studies revealed that cfGnRH-R1 had a higher affinity than cfGnRH-R2 for cGnRH-II, cfGnRH, and various other GnRH agonists. However, this was not reflected in the inositol phosphate or cAMP signal transduction properties of both types of cfGnRH-R. We therefore conclude that in catfish, functional ligand/receptor units evolved by restricted coexpression of a particular receptor in combination with a particular GnRH in particular (nearby) tissue(s).

The SCN of the hypothalamus contains a major pacemaker, which exhibits 24-h rhythms in electrical impulse frequency. Although it is known that SCN electrical activity is high during the day and low during the night, the precise relationship between electrical activity and behavioral rhythms is almost entirely unknown. The authors performed long-term recordings of SCN multiple unit activity with the aid of implanted microelectrodes in parallel with the drinking activity in freely moving mice. The animals were kept in a 12h:12h light-dark cycle (LD 12:12) and in short-day (LD 8:16) and long-day photoperiods (LD 16:8). Onsets and offsets of behavioral activity occurred when SCN discharge was around half-maximum value. Of the onsets 80%, and of the offsets 62%, occurred when SCN electrical activity differed less than 15% from the half-maximum electrical activity levels. Transitions between rest and activity could be described by a sigmoid shaped probability curve with Hill coefficients of 7.0 for onsets and 5.7 for offsets. The similarity in the onset and offset levels shows an absence of hysteresis in the control of behavioral activity by the SCN. Exposure to short- or long-day photoperiods induced significant alterations in the waveform of electrical activity but did not affect SCN electrical activity levels at which behavioral transitions occurred. In all photoperiods, the SCN signal was skewed with more rapid discharge changes during onsets (19% per hour) than offsets (11% per hour). The precision of the circadian system appears optimized, as transitions between behavioral activity and rest occur when the change in SCN electrical activity is maximal, both during the declining and rising phase. The authors conclude that transitions in behavioral state can be described by a probability function around half-maximum electrical activity levels and that the parameters of the SCN, predicting onset and offset of behavior, are remarkably insensitive to environmental conditions.

The suprachiasmatic nuclei (SCN) contain a major circadian pacemaker, which is regulated by photic and nonphotic stimuli. Although enkephalins are present in the SCN, their role in phase regulation of the pacemaker is largely unknown. The opioid agonist fentanyl, a homologue of morphine, is an addictive drug that induces phase shifts of circadian rhythms in hamsters. We observed that these phase shifts are blocked by naloxone, which is a critical test for true opioid receptor involvement, and conclude that opioid receptors are the sole mediators of the actions of fentanyl on the circadian timing system. A strong interaction between opioids and light input was shown by the ability of fentanyl and light to completely block each other's phase shifts of behavioural activity rhythms. Neuronal ensemble recordings in vitro provide first evidence that SCN cells show direct responses to fentanyl and react with a suppression of firing rate. Moreover, we show that fentanyl induces a strong attenuation of light-induced Syrian hamster Period 1 (shPer1) gene expression during the night. During the subjective day, we found no evidence for a role of shPer1 in mediation of fentanyl-induced phase shifts. Based on the present results, however, we cannot exclude the involvement of shPer2. Our data indicate that opioids can strongly modify the photic responsiveness of the circadian pacemaker and may do so via direct effects on SCN electrical activity and regulation of Per genes. This suggests that the pathways regulating addictive behaviour and the circadian clock intersect.

The mammalian central circadian pacemaker (the suprachiasmatic nucleus, SCN) contains thousands of neurons that are coupled through a complex network of interactions. In addition to the established role of the SCN in generating rhythms of ∼24 hours in many physiological functions, the SCN was recently shown to be necessary for normal self-similar/fractal organization of motor activity and heart rate over a wide range of time scales—from minutes to 24 hours. To test whether the neural network within the SCN is sufficient to generate such fractal patterns, we studied multi-unit neural activity of in vivo and in vitro SCNs in rodents. In vivo SCN-neural activity exhibited fractal patterns that are virtually identical in mice and rats and are similar to those in motor activity at time scales from minutes up to 10 hours. In addition, these patterns remained unchanged when the main afferent signal to the SCN, namely light, was removed. However, the fractal patterns of SCN-neural activity are not autonomous within the SCN as these patterns completely broke down in the isolated in vitro SCN despite persistence of circadian rhythmicity. Thus, SCN-neural activity is fractal in the intact organism and these fractal patterns require network interactions between the SCN and extra-SCN nodes. Such a fractal control network could underlie the fractal regulation observed in many physiological functions that involve the SCN, including motor control and heart rate regulation.

Trait-like individual differences in the melanopsin phototransduction circuitry contribute to individual differences in sleep timing. Blue light-sensitive young individuals are more prone to delayed sleep.

R192Q migraine mice lack the physiological retardation in circadian adaptation to phase advance shifts. The opposite findings in vivo and in vitro exclude involvement of the retinal input pathway or the phase-shifting capacity of the SCN. Thus, the physiological inhibitory process appears to be mediated by Ca(v)2.1 channel-dependent afferent signaling from extra-SCN brain areas to the SCN.