Liquid–liquid phase separation (LLPS) underlies diverse biological processes. Because most LLPS studies were performed in vitro using recombinant proteins or in cells that overexpress protein, the physiological relevance of LLPS for endogenous protein is often unclear. PERIOD, the intrinsically disordered domain-rich proteins, are central mammalian circadian clock components and interact with other clock proteins in the core circadian negative feedback loop. Different core clock proteins were previously shown to form large complexes. Circadian clock studies often rely on experiments that overexpress clock proteins. Here, we show that when Per2 transgene was stably expressed in cells, PER2 protein formed nuclear phosphorylation-dependent slow-moving LLPS condensates that recruited other clock proteins. Super-resolution microscopy of endogenous PER2, however, revealed formation of circadian-controlled, rapidly diffusing nuclear microbodies that were resistant to protein concentration changes, hexanediol treatment, and loss of phosphorylation, indicating that they are distinct from the LLPS condensates caused by protein overexpression. Surprisingly, only a small fraction of endogenous PER2 microbodies transiently interact with endogenous BMAL1 and CRY1, a conclusion that was confirmed in cells and in mice tissues, suggesting an enzyme-like mechanism in the circadian negative feedback process. Together, these results demonstrate that the dynamic interactions of core clock proteins are a key feature of mammalian circadian clock mechanism and the importance of examining endogenous proteins in LLPS and circadian clock studies.

We investigated the consequence of no night environment (constant light, LL) on reproductive performance in zebra finches in the parent (P) and subsequent F1 generation. As a measure of the overall effects on the metabolic reproductive health, we monitored daily activity behaviour, recorded song and cheek patch size in males, and measured body size and hormone levels. As compared to controls under 12 h light: 12 h darkness (12:12 h LD), both P and F1 pairs showed a compromised reproductive success, as evidenced by fewer fledglings and viable offsprings with longer fledging durations, and increased offspring mortality with successive three clutches under LL. The overall negative effect of the no-night environment was increased in the F1generation. As compared to P pairs, F1pairs had more failed nesting and breeding attempts, took longer to initiate reproduction, incubated fewer eggs, produced fewer viable offspring with longer fledging duration, and showed increased offspring mortality. Consistent with negative reproductive effects, P males showed significant changes in the motif duration and other spectral features of song, and both F1 and F2 males copied poorly the song of their parent under LL. Plasma corticosterone and sex hormones (testosterone in males and estradiol in females) levels were significantly lower under LL. Daily plasma melatonin rhythm persisted but with a reduced amplitude under LL. These results demonstrate the importance of night in reproduction in a continuously breeding diurnal species, and give insights into the possible impact on physiology of animals whose surrounding environment is consistently losing the darkness of night.

The last 50 years have witnessed extraordinary discoveries in the field of circadian rhythms. However, there are still several mysteries that remain. One of these chronobiological mysteries is the circadian rhythm that is revealed by administration of stimulant drugs to rodents. Herein we describe the discovery of this circadian rhythm and its underlying oscillator, which is frequently called the methamphetamine-sensitive circadian oscillator, or MASCO. This oscillator is distinct from canonical circadian oscillators because it controls robust activity rhythms independently of the suprachiasmatic nucleus and circadian genes are not essential for its timekeeping. We discuss these fundamental properties of MASCO and integrate studies of strain, sex, and circadian gene mutations on MASCO. The anatomical loci of MASCO are not known, so it has not been possible thus far to discover its novel molecular timekeeping mechanism or its functional significance. However, studies in mutant mice suggest that genetic approaches can be used to identify the neural network involved in the rhythm generation of MASCO. We also discuss parallels between human and rodent studies that support our working hypothesis that a function of MASCO may be to regulate sleep-wake cycles.

In recent times, there has been an unprecedented increase in usage of electrical lightning. This has led to increase in artificial light at night (ALAN), and it has been suggested as a source of environmental pollution. ALAN exposure has been reported to be associated with disruption of daily rhythms and serious health consequences, such as immune, metabolic, and cognitive dysfunctions in both birds and mammals. Given the worldwide pervasiveness of ALAN, this research topic is also important from an ecological perspective. In birds, daily timings and appropriate temporal niches are important for fitness and survival. Daily rhythms in a wide array of functions are regulated by the circadian clock(s) and endogenous oscillators present in the body. There is accumulating evidence that exposure to ALAN disrupts clock-regulated daily rhythms and suppresses melatonin and sleep in birds. Circadian clock, melatonin, and sleep regulate avian cognitive performance. However, there is limited research on this topic, and most of the insights on the adverse effects of ALAN on cognitive functions are from behavioural studies. Nevertheless, these results raise an intriguing question about the molecular underpinning of the ALAN-induced negative consequences on brain functions. Further research should be focused on the molecular links between ALAN and cognitive performance, including the role of melatonin, which could shed light on the mechanism by which ALAN exposures lead to negative consequences.