Circadian nature of the photoperiodic clock in Japanese quail

Summary The photoperiodic clock in quail (Coturnix colurnix japonica) is based upon a rhythm of photoinducibility (Øi) but the extent to which this rhythm is circadian remains unclear. Two types of experiment investigated this situation. In the first, gonadectomized quail were adapted to live in periods of darkness by training them on a schedule containing one short day and 3 days of darkness (SD/DD/DD/DD). They were then exposed to a single pulse of 6 or 10 h of light at different times across 3 days of darkness. The photoperiodic response, measured by the increase in LH secretion, showed clear rhythmicity, demonstrating unequivocally the circadian nature of Øi. The second set of experiments employed Nanda-Hamner cycles and varied the length of the photoperiod from 6 to 11 h. Responsiveness in a 36 h or a 60 h cycle was highly dependent upon the length of the photoperiod, something not predicted from theory. For instance, LD 6:30 was not photoperiodically inductive but LD 10:26 was clearly inductive. Close analysis of patterns of LH secretion indicated an unexpected delay before induction occurred and then a rapid rise to a stable level of induction. When LH was measured in every pulse under LD 10:26 there was no evidence that LH levels alternately increased and decreased. This is not consistent with the simplest interpretation of Nanda-Hamner experiments where alternate pulses of light are thought to entrain the rhythm or induce a photoperiodic response by coinciding with Øi. It is concluded that the quail's photoinducible rhythm is indeed based on a circadian rhythm but one that is only weakly self-sustaining. Possibly as a consequence of this, the rhythm's behaviour under abnormal photoperiodic cycles may be rather different from that found in other species and from other circadian rhythms in quail.Abbreviations Øi photoinducible phase - LH luteinizing hormone     

Circadian clock adjustment to plant iron status depends on chloroplast and phytochrome function

Plant chloroplasts are not only the main cellular location for storage of elemental iron (Fe), but also the main site for Fe, which is incorporated into chlorophyll, haem and the photosynthetic machinery. How plants measure internal Fe levels is unknown. We describe here a new Fe‐dependent response, a change in the period of the circadian clock. In Arabidopsis, the period lengthens when Fe becomes limiting, and gradually shortens as external Fe levels increase. Etiolated seedlings or light‐grown plants treated with plastid translation inhibitors do not respond to changes in Fe supply, pointing to developed chloroplasts as central hubs for circadian Fe sensing. Phytochrome‐deficient mutants maintain a short period even under Fe deficiency, stressing the role of early light signalling in coupling the clock to Fe responses. Further mutant and pharmacological analyses suggest that known players in plastid‐to‐nucleus signalling do not directly participate in Fe sensing. We propose that the sensor governing circadian Fe responses defines a new retrograde pathway that involves a plastid‐encoded protein that depends on phytochromes and the functional state of chloroplasts.     

The circadian clock contributes to diurnal patterns of plant indirect defense in nature

The plant circadian clock regulates the rhythms of plant metabolism. Many herbivore-induced plant volatiles(HIPVs) fluctuate, diurnally, but the role of the circadian clock in the emission of HIPVs and their ecological consequences remains largely unknown.Here, we show that the timing of herbivore attack can alter the outcome of tri-trophic interactions, and this is mediated by the circadian clock, under both field and glasshouse conditions. Although most HIPV emissions did not have a circadian rhythm, the circadian clock modulated HIPV emissions in a time-dependent manner. HIPVs mediate tri-trophic interactions, and the circadian clock may affect these interactions by modulating HIPV emission in nature.     

Guard cells in albino leaf patches do not respond to photosynthetically active radiation, but are sensitive to blue light, CO2 and abscisic acid

Stomatal openings can be stimulated by light through two signalling pathways. The first pathway is blue light specific and involves phototropins, while the second pathway mediates a response to photosynthetically active radiation (PAR). This second pathway was studied with the use of albino Vicia faba plants and variegated leaves of Chlorophytum comosum. Treatment of V. faba with norflurazon (Nf) inhibits the synthesis of carotenoids and leads to albino leaves with guard cells that lack functional green chloroplasts. Guard cells in albino leaf patches of C. comosum, however, do contain photosynthetically active chloroplasts. Stomata in albino leaf patches of both plants did not respond to red light, although blue light could still induce stomatal opening. This shows that the response to PAR is not functioning in albino leaf patches, even though guard cells of C. comosum harbour chloroplasts. Stomata of Nf-treated plants still responded to CO2 and abscisic acid (ABA). The size of Nf-treated guard cells was increased, but impalement studies with double-barrelled microelectrodes revealed no changes in ion-transport properties at the plasma membrane of guard cells. Blue light could hyperpolarize albino guard cells by triggering outward currents with peak values of 37 pA in albino plants and 51 pA in green control cells. Because of the inhibition of carotenoid biosynthesis, Nf-treated V. faba plants contained only 4% of the ABA content found in green control plants. The ABA dose dependence of anion channel activation in guard cells was shifted in these plants, causing a reduced response to 10 microM ABA. These data show that despite the dramatic changes in physiology caused by Nf, the gross responsiveness of guard cells to blue light, CO2 and ABA remains unaltered. Stomata in albino leaf patches, however, do not respond to PAR, but require photosynthetically active mesophyll cells for this response.     

Circadian rhythms in Neurospora crassa: Dynamics of the clock component frequency visualized using a fluorescent reporter

The frequency (frq) gene of Neurospora crassa has long been considered essential to the function of this organism’s circadian rhythm. Increasingly, deciphering the coupling of core oscillator genes such as frq to the output pathways of the circadian rhythm has become a major focus of circadian research. To address this coupling it is critical to have a reporter of circadian activity that can deliver high resolution spatial and temporal information about the dynamics of core oscillatory proteins such as FRQ. However, due to the difficulty of studying the expression of circadian rhythm genes in aerobic N. crassa cultures, little is known about the dynamics of this gene under physiologically realistic conditions. To address these issues we report a fluorescent fusion to the frq gene using a codon optimized version of the mCherry gene. To trace the expression and accumulation of FRQ–mCherryNC (FRQ–mCh) during the circadian rhythm, growing vegetative hyphae were scanned every hour under confocal microscopy (100×). Fluorescence of FRQ–mCh was detected only at the growing edge of the colony, and located in the cytoplasm and nuclei of vegetative hyphae for a distance of approximately 150–200 μm from the apices of leading hyphae. When driven by the frq promoter, apparently there was also a second FRQ entrance into the nucleus during the circadian cycle; however the second entrance had a lower accumulation level than the first entrance. Thus this fluorescent fusion protein has proven useful in tracking the spatial dynamics of the frq protein and has indicated that the dynamics of the FRQ protein’s nuclear trafficking may be more complex than previously realized.     

The adaptive nature of the plant circadian clock in natural environments

The plant circadian clock coordinates developmental, physiological, and metabolic processes with diel changes in light and temperature throughout the year. The balance between the persistence and plasticity of the clock in response to predictable and unpredictable environmental changes may be key to the clock’s adaptive nature across temporal and spatial scales. Studies under controlled conditions have uncovered critical signaling pathways involved in light and temperature perception by the clock; however, they don’t account for the natural lag of temperature behind photoperiod. Studies in natural environments provide key insights into the clock’s adaptive advantage under more complex natural settings. Here, we discuss the role of the circadian clock in light and temperature perception and signaling, how the clock integrates these signals for a coordinated and adaptive response, and the adaptive advantage conferred by the clock across time and space in natural environments.

The circadian clock orchestrates the precise coordination of plant processes across daily and annual cycles of light and temperature, resulting in adaptation to local environments.     

Circadian variations in self-monitoring,a component of executive functions

The objective of this study was to identify circadian rhythms in self-monitoring, a component of executive functions. Participants were 10 undergraduate students, age: 18.5 ± 2.68 years, two male and eight female. They were recorded on a 30-h constant routine protocol; rectal temperature was recorded every minute and performance on a tracking task was assessed every 100 min. Self-monitoring indicators were adjustments of responses to random changes of speed and trajectory of a circle moving on the computer screen. Participants showed better accuracy during the afternoon, with decreases in the morning (06:20 and 08:00 h). These variations showed a phase delay of 2:29 ± 2:19 h with respect to the circadian rhythm of body temperature. In conclusion, there are circadian variations in self-monitoring. The decline in this component of executive functions could cause serious accidents among people working or studying during a morning shift, as well as commuting to and from work or school.     

Cells respond to and bind countin,a component of a multisubunit cell number counting factor

In Dictyostelium discoideum counting factor (CF), a secreted approximately 450-kDa complex of polypeptides, inhibits group and fruiting body size. When the gene encoding countin (a component of CF) was disrupted, cells formed large groups. We find that recombinant countin causes developing cells to form small groups, with an EC(50) of approximately 3 ng/ml, and affects cAMP signal transduction in the same manner as semipurified CF. Recombinant countin increases cell motility, decreases cell-cell adhesion, and regulates gene expression in a manner similar to the effect of CF. However, countin does not decrease adhesion or group size to the extent that semipurified CF does. A 1-min exposure of developing cells to countin causes an increase in F-actin polymerization and myosin phosphorylation and a decrease in myosin polymerization, suggesting that countin activates a rapid signal transduction pathway. (125)I-Labeled countin has countin bioactivity, and binding experiments suggest that vegetative and developing cells have approximately 53 cell-surface sites that bind countin with a K(D) of approximately 1.5 ng/ml or 60 pm. We hypothesize that countin regulates cell development through the same pathway as CF and that other proteins within the complex may modify the activity of countin and/or have independent size-regulating activities.     

Circadian clock functioning is linked to acute stress reactivity in rats

At least two major physiological systems are involved in the adaptation of the organism to environmental challenges: the circadian system and the stress reaction. This study addressed the possibility that interindividual differences in stress sensitivity and in the functioning of the circadian system are related. At 2 months of age, corticosterone secretion in response to a 20-min restraint stress was assessed in 9 Sprague-Dawley rats for which running wheel activity was recorded as a rhythmic behavioral marker of the circadian clock. Two weeks later, the adaptive response of the circadian system to an abrupt shift in the light:dark (LD) cycle was assessed in those rats using a jet-lag paradigm. Finally, after resynchronization to the new LD cycle, rats were transferred to constant darkness to assess the free-running period of their circadian rhythm of running-wheel activity. Results indicate that stress-induced corticosterone secretion was (1) positively correlated with the number of days to resynchronize the circadian activity rhythm to the new LD cycle, and with the value of its free-running period, and (2) negatively correlated with the intensity of daily locomotor activity. Those data, emphasizing the interactions between the stress response of an organism and the functioning of its circadian system, could explain interindividual differences in humans' susceptibility to shift work or other circadian-related disorders.     

Circadian phototransduction: phase resetting and frequency of the circadian clock of Gonyaulax cells in red light

Constant red light (RR) influences the Gonyaulax clock in several ways: (1) Phase resetting by white or blue light pulses is stronger under background RR than in constant white light (WW); (2) frequency of the rhythm is less in RR than in WW; and (3) the amplitude of the spontaneous flashing rhythm is greater in RR than in WW. The phase response curve (PRC) to 4-hr white or blue light pulses is of high amplitude (Type 0) for cells in RR, but is of lower amplitude (Type 1) for cells in WW. In all cases, the PRC is highly asymmetrical: The magnitude of advance phase resetting is far higher than that of delay resetting. Consistent with this PRC, Gonyaulax cells in RR (free-running period greater than 24 hr) will entrain to T cycles of between 21 and 26.5 hr. The bioluminescence rhythms exhibit "masking" by blue light pulses while entrained to these T cycles. The fluence response of phase resetting to light-pulse intensity is not linear or logarithmic--rather, it is discontinuous. This feature is consistent with a limit cycle interpretation of Type 0 resetting of circadian clocks. Light pulses that cause large phase shifts also shorten the subsequent free-running period. The phase angle difference between the clock and the previous LD cycle is within 2 hr of the same phase between 16 degrees C and 25 degrees C, as determined from the light PRCs at various temperatures. Several drugs that inhibit mitochondria and/or electron transport will partially inhibit the phase shift by light.     

Circadian timing of cricket calling song: a clock persisting from nymphs to adults

ABSTRACT. Male Australian field crickets (Teleogryllus commodus , Walker) reared in LD 12:12 h were transferred to LL at different developmental stages and the timing of their circadian calling song rhythm was analysed in regard to the previous zeitgeber. The phase settings for the onset and end of activity were similar in crickets experiencing the LD/LL transition: (i) 3–52 days after the final moult, (ii) within 24 h before the final moult, or (iii) 1–10 days before the final moult. For all groups the results reveal entrainment of the circadian mechanism at the last LD, thus excluding age-related differences. The rhythms of crickets, transferred from LD to LL as larval instars and also exposed to a reduced temperature (5–8^oC) during their last night, were delayed by about 11 h, an effect similar to that in adult crickets after a comparable cold exposure (Loher & Wiedenmann, 1981).
The results are interpreted showing that the circadian control of (the adult's) calling song already functions in the previous (non-singing) larval stages. Since the rhythmicity continued through moults and sexual maturation, it is concluded that the control centres regulating those physiological processes (e.g. pars intercerebralis, corpora allata) are not essential to the basic circadian mechanism.