Biological Sex Influences Daily Locomotor Rhythms in Mice Held Under Different Housing Conditions.

Daily rhythms are programmed by a central circadian clock that is modulated by photoperiod. Here, we recorded locomotor activity rhythms in C57Bl/6 or mPer2Luc mice of both sexes held under different housing conditions. First, we confirm that the structure of locomotor activity rhythms differs between male and female mice in both genetic backgrounds. Male mice exhibit a nightly "siesta," whereas female mice fluctuate between nights with and without a nightly siesta, which corresponds with changes in locomotor activity levels, circadian period, and vaginal cytology. The nightly siesta is modulated by the presence of a running wheel in both sexes but is not required for the infradian patterning of locomotor rhythms in females. Finally, photoperiodic changes in locomotor rhythms differed by sex, and females displayed phase-jumping responses earlier than males under a parametric photoentrainment assay simulating increasing day length. Collectively, these results highlight that sex and sex hormones influence daily locomotor rhythms under a variety of different environmental conditions.

Somatostatin regulates central clock function and circadian responses to light.

Daily and annual changes in light are processed by central clock circuits that control the timing of behavior and physiology. The suprachiasmatic nucleus (SCN) in the anterior hypothalamus processes daily photic inputs and encodes changes in day length (i.e., photoperiod), but the SCN circuits that regulate circadian and photoperiodic responses to light remain unclear. Somatostatin (SST) expression in the hypothalamus is modulated by photoperiod, but the role of SST in SCN responses to light has not been examined. Our results indicate that SST signaling regulates daily rhythms in behavior and SCN function in a manner influenced by sex. First, we use cell-fate mapping to provide evidence that SST in the SCN is regulated by light via de novo Sst activation. Next, we demonstrate that Sst  -/- mice display enhanced circadian responses to light, with increased behavioral plasticity to photoperiod, jetlag, and constant light conditions. Notably, lack of Sst  -/- eliminated sex differences in photic responses due to increased plasticity in males, suggesting that SST interacts with clock circuits that process light differently in each sex. Sst  -/- mice also displayed an increase in the number of retinorecipient neurons in the SCN core, which express a type of SST receptor capable of resetting the molecular clock. Last, we show that lack of SST signaling modulates central clock function by influencing SCN photoperiodic encoding, network after-effects, and intercellular synchrony in a sex-specific manner. Collectively, these results provide insight into peptide signaling mechanisms that regulate central clock function and its response to light.

Sex differences in daily timekeeping and circadian clock circuits.

The circadian system regulates behavior and physiology in many ways important for health. Circadian rhythms are expressed by nearly every cell in the body, and this large system is coordinated by a central clock in the suprachiasmatic nucleus (SCN). Sex differences in daily rhythms are evident in humans and understanding how circadian function is modulated by biological sex is an important goal. This review highlights work examining effects of sex and gonadal hormones on daily rhythms, with a focus on behavior and SCN circuitry in animal models commonly used in pre-clinical studies. Many questions remain in this area of the field, which would benefit from further work investigating this topic.

Photoperiod Manipulation Reveals a Light-Driven Component to Daily Patterns of Ventilation in Male C57Bl/6J Mice.

Obstructive sleep apnea is a common sleep disorder that increases risk for cardiovascular disease and mortality. The severity of sleep-disordered breathing in obstructive sleep apnea patients fluctuates with the seasons, opening the possibility that seasonal changes in light duration, or photoperiod, can influence mechanisms of breathing. Photoperiod can have profound effects on internal timekeeping and can reshape metabolic rhythms in mammals. While the daily rhythm in ventilation is largely shaped by the metabolic rate, less is known about whether ventilatory rhythms are altered in accordance with metabolism under different photoperiods. Here, we investigate the relationship between ventilation and metabolism under different photoperiods using whole-body plethysmography and indirect calorimetry. We find that the daily timing of ventilation is chiefly synchronized to dark onset and that light cues are important for maintaining daily ventilatory rhythms. Moreover, changes in ventilatory patterns are not paralleled by changes in oxygen consumption, energy expenditure, or respiratory exchange rate under different photoperiods. We conclude that ventilatory patterns are not only shaped by the metabolic rate and circadian timing but are also influenced by other light-driven factors. Collectively, these findings have clinical implications for the seasonal variations in sleep-disordered breathing found in individuals with obstructive sleep apnea.

Marking Time: Colorful New Insights into Master Clock Circuits.

A neural clock controls what we do each day, and understanding its circuitry is important for health. In this issue of Neuron, Shan et al. visualize molecular rhythms in subtypes of master clock neurons to test principles of cell identity and network wiring.

Reduced VIP Expression Affects Circadian Clock Function in VIP-IRES-CRE Mice (JAX 010908).

Circadian rhythms are programmed by the suprachiasmatic nucleus (SCN), which relies on neuropeptide signaling to maintain daily timekeeping. Vasoactive intestinal polypeptide (VIP) is critical for SCN function, but the precise role of VIP neurons in SCN circuits is not fully established. To interrogate their contribution to SCN circuits, VIP neurons can be manipulated specifically using the DNA-editing enzyme Cre recombinase. Although the Cre transgene is assumed to be inert by itself, we find that VIP expression is reduced in both heterozygous and homozygous adult VIP-IRES-Cre mice (JAX 010908). Compared with wild-type mice, homozygous VIP-Cre mice display faster reentrainment and shorter free-running period but do not become arrhythmic in constant darkness. Consistent with this phenotype, homozygous VIP-Cre mice display intact SCN PER2::LUC rhythms, albeit with altered period and network organization. We present evidence that the ability to sustain molecular rhythms in the VIP-Cre SCN is not due to residual VIP signaling; rather, arginine vasopressin signaling helps to sustain SCN function at both intracellular and intercellular levels in this model. This work establishes that the VIP-IRES-Cre transgene interferes with VIP expression but that loss of VIP can be mitigated by other neuropeptide signals to help sustain SCN function. Our findings have implications for studies employing this transgenic model and provide novel insight into neuropeptide signals that sustain daily timekeeping in the master clock.

Circuit development in the master clock network of mammals.

Daily rhythms are generated by the circadian timekeeping system, which is orchestrated by the master circadian clock in the suprachiasmatic nucleus (SCN) of mammals. Circadian timekeeping is endogenous and does not require exposure to external cues during development. Nevertheless, the circadian system is not fully formed at birth in many mammalian species and it is important to understand how SCN development can affect the function of the circadian system in adulthood. The purpose of the current review is to discuss the ontogeny of cellular and circuit function in the SCN, with a focus on work performed in model rodent species (i.e., mouse, rat, and hamster). Particular emphasis is placed on the spatial and temporal patterns of SCN development that may contribute to the function of the master clock during adulthood. Additional work aimed at decoding the mechanisms that guide circadian development is expected to provide a solid foundation upon which to better understand the sources and factors contributing to aberrant maturation of clock function.

Physiological, behavioral and environmental factors influence bifurcated circadian entrainment in mice.

Under permissive conditions, mice and hamsters exposed to a polyphasic light regime consisting of two light and two dark phases every 24 h (Light:Dark:Light:Dark; LDLD) can adopt a bifurcated entrainment pattern with roughly equal amounts of running wheel activity in each of the two nights. This rhythm "bifurcation" has significant after-effects on increased circadian adaptability: Mice that have been bifurcated show accelerated rates of re-entrainment after a sudden phase shift and have a markedly expanded range of entrainment. Identifying environmental and physiological factors that facilitate or prevent rhythm bifurcation in LDLD conditions will contribute to an understanding of mechanisms underlying enhanced circadian plasticity. Here we investigate the effects of sex, age, light intensity, access to a running wheel, melatonin, and diet composition on bifurcation behaviors of mice (C57Bl/6 J) exposed to LDLD. Female mice and young mice (<20 weeks) express more symmetrically bifurcated activity compared to male mice and older mice (>30 weeks). Additionally and independently, higher photophase intensities (~500 lx) predict more symmetric entrainment than low levels of light (~50 lx). Without access to a functional running-wheel, mice do not adopt bimodal activity patterns and only transiently maintain them, suggesting that high levels of aerobic activity are necessary for rhythm bifurcation. Neither a lifetime exposure to melatonin administered in the drinking water nor a high fat diet affected bifurcation. Collectively, these results demonstrate that circadian plasticity can be strongly modulated by intrinsic and extrinsic factors. With enhanced mechanistic understanding of this modulation, it may be possible to render human clocks more adaptable and thereby ameliorate negative consequences associated with repeated jet-lag or shift-work.

Photoperiodic Requirements for Induction and Maintenance of Rhythm Bifurcation and Extraordinary Entrainment in Male Mice.

Exposure of mice to a 24 h light:dark:light:dark (LDLD) cycle with dimly illuminated nights induces the circadian timing system to program two intervals of activity and two intervals of rest per 24 h cycle and subsequently allows entrainment to a variety of extraordinary light regimens including 30 h LDLD cycles. Little is known about critical lighting requirements to induce and maintain this non-standard entrainment pattern, termed "bifurcation," and to enhance the range of apparent entrainment. The current study determined the necessary duration of the photophase for animals to bifurcate and assessed whether requirements for maintenance differed from those for induction. An objective index of bifurcated entrainment varied with length of the photophase over 4-10 h durations, with highest values at 8 h. To assess photic requirements for the maintenance of bifurcation, mice from each group were subsequently exposed to the LDLD cycle with 4 h photophases. While insufficient to induce bifurcation, this photoperiod maintained bifurcation in mice transferred from inductive LDLD cycles. Entrainment to 30 h LDLD cycles also varied with photoperiod duration. These studies characterize non-invasive tools that reveal latent flexibility in the circadian control of rest/activity cycles with important translational potential for addressing needs of human shift-workers.