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This study investigated the relative potency of melatonin and arousal as Zeitgebers in the non-photic phase shifting of circadian rhythmicity in the adult Syrian hamster. Animals held under dim red light (DD) exhibited robust free-running rhythms of wheel-running activity. Melatonin (1 mg/kg) or ethanolic saline vehicle, delivered manually by subcutaneous injection after removing the animal from its cage, resulted in phase advances of the activity rhythm. This effect was phase dependent, injections at CT 8 and 10 being effective (CT 12 = anticipated activity onset), whereas injection at CT 2, 6, 14 and 20 did not cause a shift. There was no significant difference between the magnitude or timing of phase shifts in response to injections of saline or melatonin. To determine whether the observed shifts were related to arousal of the animals induced by handling, a second group held under DD were fitted with chronic s.c. cannulae so that melatonin solution or vehicle could be delivered remotely at projected CT 10. Neither solution had any effect upon the free-running rhythm. However, when these animals received manual s.c. injection of saline or melatonin solution, they exhibited phase advances similar to those observed in Expt. 1. These results fail to support the hypothesis that melatonin can exert a chemically specific, acute phase-shifting action in the adult Syrian hamster. They do, however, demonstrate the potent effect of arousing stimuli upon the circadian clock in this species.

Single episodes of arousal of Syrian hamsters 2 h before projected activity onset (i.e., CT 10) phase-advanced their free-running circadian rhythm of wheel-running. Serial arousal once every 23 h or once every 23.5 h for 7 days caused large composite phase-advances to the wheel-running rhythm, the latter period being more effective in supporting an interval of stable entrainment. Pre-treatment of hamsters at CT 6 with the serotonergic antagonist ritanserin (1–5 mg/kg), which acts at both 5-HT 2 and the putative 5-HT 7 receptor, impaired the phase-advancing response to arousal at CT 10 but the drug was without effect on phase advances induced by exposure to light. Pre-treatment with a second serotonergic antagonist, ketanserin (1–5 mg/kg), which is without effect at 5-HT 7 but has high affinity for 5-HT 2 receptors, was also effective in attenuating the phase advancing effect of arousal at CT 10. However, neither agent was able to achieve complete blockade of the phase advances. These results are discussed in relation to in vitro and in vivo studies in the rat which have identified a role for 5-HT 7 receptors in serotonin-mediated circadian entrainment.

Photic entrainment of the circadian oscillator located in the hypothalamic suprachiasmatic nucleus (SCN) is considered to be mediated at least partly by release of glutamate from the retinal presynaptic nerve terminals acting via a NMDA receptor. Several NMDA receptor subtypes have been cloned and expressed in model systems. The NMDA-R1 subtype is essential for the function of the NMDA receptor, and the multiple NMDA-R2(-A, -B, or -C) sububits potentiate and differentiate the function of the NMDA receptor by forming different heteromeric configuration with NMDA-R1. The aim of this study was to use in situ hybridization histochemistry with oligonucleotide sequences (42–48-mer) labeled with 35S to detect whether NMDA receptor mRNA is present in the rat SCN, and if so, to characterize which receptor subtypes occur. In order to identify the precise location of NMDA receptor mRNAs within the SCN, sections were dipped in emulsion and cellular resolution was achieved. The hybridization revealed a high abundance of NMDA-R1 mRNA in the SCN as well as in many other forebrain areas. The NMDA-R1 expressing cells were distributed throughout the SCN. NMDA-R2A and NMDA-R2B mRNAs were found in the hippocampus, but not in the SCN. In contrast, NMDA-R2C mRNA was found in relative high amounts in the rat SCN, but not in other hypothalamic areas. In dipped sections, it was evident that the localization of NMDA-2RC was mostly confined to the dorsomedial part of the SCN. Thus, the rat SCN contains a specific combination of NMDA receptor mRNA subtypes not found in other forebrain structures. These observations are consistent with the hypothesis that glutamate mediates the effect of light on entrainment of the circadian oscillator.