Journal of Biological Rhythms

The daily rhythm of 24 h and the annual rhythm of 12 mo constitute the 2 major, highly predictable rhythms of the biosphere. The internal circadian clock enables organisms to track daily changes in their environment; the photoperiodic timer, alone or in concert with a circannual clock, enables organisms to anticipate and prepare in advance for seasonal changes in their environment. The circadian clock entrains to dawn and dusk and tracks light and temperature on a day-to-day basis, while the photoperiodic timer serves as a long-term, physiological go/no-go switch that commits an animal to development, reproduction, dormancy, or migration on a seasonal or even lifetime basis. In 1936, Erwin Bünning proposed that circadian rhythms formed the basis (Grundlage) for photoperiodic response to day length. Historical inertia generated by correlative evidence from early physiological studies and a proliferating number of descriptive models has resulted in the widespread assumption that the circadian clock constitutes the necessary, causal basis of photoperiodism in general. This historical inertia has also restricted the search for genes involved in insect photoperiodism to genes central to the circadian clock in Drosophila and has led investigators to conclude that any behavior, process, or gene expression that varies with day length represents photoperiodism or a gene involved in photoperiodism. The authors discuss how blinders imposed by the circadian imperative have retarded progress toward identifying the genetic basis of photoperiodism and how the insights gained from geographic variation in photoperiodic response have been used to show the independent evolution of the circadian clock and photoperiodism. When geographic variation is found in circadian genes, the most immediate and parsimonious search for adaptive significance should be in circadian function, not in extrapolation to photoperiodism. Finally, the authors propose that circadian-unbiased, forward genetic approaches should be used to identify genes involved in photoperiodism within extant populations and among populations over evolutionary time.

During the hibernation season, mammalian hibernators alternate between prolonged bouts of torpor with a reduced body temperature (Tb) and short arousals with a return to euthermy. Evidence is presented here to show that this metabolic—and also physiological and neuroanatomical—rhythm is controlled by a clock, the torpor-arousal (TA) clock. The temperature dependence of torpor bout duration in 3 species of Spermophilus (published data) may be described by assuming that the TA clock is a circadian clock (probably not the suprachiasmatic clock) that has lost its temperature compensation. This loss might result either from a permanent deletion, or more likely from a seasonal epigenetic control at the level of the clock gene machinery. This hypothesis was verified over the full Tb range on published data from 5 other species (a monotreme, a marsupial, and 3 placental mammals). In a hibernation season, instantaneous subjective time of the putative TA clock was summated over each torpor bout. For each animal, torpor bout length (TBL) was accurately predicted as a constant fraction of a subjective day, for actual durations in astronomical time varying between 4 and 13 to 20 days. The resulting temperature dependence of the interval between arousals predicts that energy expenditure over the hibernation season will be minimal when Tb is as low as possible without eliciting cold thermogenesis.

To conserve scarce energetic resources during winter, seasonal breeders inhibit reproduction and other nonessential behavioral and physiological processes. Reproductive cessation is initiated in response to declining day lengths, a stimulus represented centrally as a long-duration melatonin signal. The melatonin signal is not decoded by the reproductive axis directly, but by an unidentified neurochemical system upstream of gonadotropin-releasing hormone (GnRH). The dorsomedial nucleus of the hypothalamus (DMH) has been implicated in seasonal changes in reproductive function in Syrian hamsters (Mesocricetus auratus), although the specific-cell phenotype decoding photoperiodic information remains unknown. RFamide-related peptide (RFRP; the mammalian homolog of the gonadotropin-inhibitory hormone (GnIH) gene identified in birds) has emerged as a potent inhibitory regulator of the reproductive axis and, significantly, its expression is localized to cell bodies of the DMH in rodents. In the present study, the authors explored the relationship between RFRP expression, photoperiod exposure, and reproductive condition/hormonal status. In male hamsters that respond to short days with reproductive inhibition, RFRP-ir and mRNA expression are markedly reduced relative to long-day animals. Replacement of testosterone in short-day animals did not affect this response, suggesting that alterations in RFRP expression are not a result of changing sex steroid concentrations. A subset of the hamster population that ignores day length cues and remains reproductively competent in short days (nonresponders) exhibits RFRP-ir expression comparable to long-day hamsters. Analysis of cell body and fiber density suggests a potential interplay between peptide production and release rate in differentially regulating the reproductive axis during early and late stages of reproductive regression. Together, the present findings indicate that photoperiod-induced suppression of reproduction is associated with changes in RFRP and mRNA expression, providing opportunity for further exploration on the role that RFRP plays in this process.

The mammalian endogenous circadian clock, the suprachiasmatic nuclei, receives environmental inputs, namely the light-dark cycle, through photopigments located in the eye and from melanopsin-expressing retinal ganglion cells. The authors investigated the influence of light wavelength and intensity on the synchronization of the rest-activity rhythm of the gray mouse lemur, a nocturnal Malagasy primate. Animals were tested at different irradiance levels (320, 45, 13, and 6 nmol.m^-2.s^{- 1}) under several light wavelengths (from 400 to 610 nm). Several parameters including circadian period, activity, and body temperature waveforms were used to assess synchronization to a 12:12 light-dark cycle in comparison to control treatments (12:12 white light or continuous darkness). Entrainment of the circadian rest-activity cycle increased with light intensity. It was more efficient for mid wavelengths relative to shorter or longer wavelengths but not coincident with melanopsin maximal sensitivity, suggesting other photoreceptors are likely involved in lemurs’ photoentrainment. The authors obtained a novel synchronization pattern characterized by a clear synchronization to lights-on only without phasing to lights-off. Changes in photo-responsiveness at dusk and dawn highlight differential responses of evening and morning oscillators in the circadian clock.

In nocturnal rodents, millisecond light (“flash”) stimuli can induce both a large circadian rhythm phase shift and an associated state change from highly active to quiescence followed by behavioral sleep. Suppression of locomotion (“negative masking”) is an easily measured correlate of the state change. The present mouse studies used both flashes and longer light stimuli (“pulses”) to distinguish initiation from maintenance effects of light on locomotor suppression and to determine whether the locomotor suppression exhibits temporal integration as is thought to be characteristic of phase shift responses to pulse, but not flash, stimuli. In experiment 1, locomotor suppression increased with irradiance (0.01-100 µW/cm ²), in accordance with previous reports. It also increased with stimulus duration (3-3000 sec), but interpretation of this result is complicated by the ability of light to both initiate and maintain locomotor suppression. In experiment 2, an irradiance response curve was determined using a stimulus series of 10 flashes, 2 msec each, with total flash energy varying from 0.0025 to 110.0 J/m². This included a test for temporal integration in which the effects of two equal energy series of flashes that differed in the number of flashes per series (10 vs 100), were compared. The 10 flash series more effectively elicited locomotor suppression than the 100 flash series, a result consistent with prior observations involving flash-induced phase shifts. In experiment 3, exposure of mice to an 11-h light stimulus yielded irradiance-dependent locomotor suppression that was maintained for the entire stimulus duration by a 100-µW/cm² stimulus. Light has the ability to initiate a time-limited (30-40 min) interval of locomotor suppression (initiation effect) that can be extended by additional light (maintenance effect). Temporal integration resembling that seen in phase-shifting responses to light does not exist for either phase shift or locomotor suppression responses to flashes or for locomotor suppression responses to light pulses. The authors present an alternative interpretation of data thought to demonstrate temporal integration in the regulation of phase shift responses to light pulses.

Multisynaptic neural and endocrine pathways from the suprachiasmatic nucleus of the hypothalamus have been hypothesized to communicate circadian and photic information to the adrenal glands. In humans, light exposure has been reported to have no effect, increase, or decrease cortisol levels. These inconsistent findings in humans may be related to differences among studies including the intensity (~500 to 5500 lux), duration (15 min to 4 h), and circadian phase of light exposure. The authors assessed the influence of exposure to bright light on cortisol levels in humans during the rising and descending phases of the circadian rhythm of cortisol, that is, when cortisol levels are high. Twenty healthy men and women were studied using a within-subject research design. Subjects were studied in an environment free of time cues for 9 to 10 days. Subjects received a 6.7-h exposure of bright light (~10,000 lux; equivalent to ambient light intensity just after sunrise or just before sunset) or dim light (~3 lux; equivalent to candlelight) during the biological night and morning. Bright light exposure significantly reduced plasma cortisol levels at both circadian phases studied, whereas dim light exposure had little effect on cortisol levels. The finding of an acute suppressive effect of bright light exposure on cortisol levels supports the existence of a mechanism by which photic information can acutely influence the human adrenal glands.

The physiological mechanisms underlying interindividual differences in chronotype have yet to be established, although evidence suggests both circadian and homeostatic processes are involved. A physiologically based model is developed by combining models of the sleep-wake switch and circadian pacemaker, providing a means of examining how interactions between these systems affect chronotype. Specifically, chronotype is shown to depend on the relative influences of homeostatic and circadian drives, with a stronger homeostatic drive causing morningness. Changes to intrinsic circadian and homeostatic properties, including homeostatic clearance and production rates, and circadian period and amplitude, are also shown to affect chronotype. These results provide a framework for explaining several experimentally observed phenomena, including age-related morningness, adolescent eveningness, and familial advanced and delayed sleep-phase disorders. Additionally, experimental studies have shown that healthy adults on the extremes of the morningness-eveningness spectrum fall into two subtypes: those whose circadian phase markers are unaffected by chronotype, and those whose circadian phase markers track their chronotype. The model demonstrates that this spectrum likely results from interindividual differences in homeostatic kinetics in the first group, and differences in circadian period in the second group. Physiologically based modeling can thus guide diagnosis of sleep pathologies.

The use of luciferase reporter genes together with luminescence detection has enabled high frequency monitoring of molecular circadian clock function in living tissues. With the help of an intensified CCD camera combined with an inverted epifluorescence microscope, the authors have established a new imaging strategy that makes use of transgenic cell type-specific expression of fluorescent proteins to identify cells of interest for subsequent circadian luminescence recording at single-cell resolution.