Evolution and population biology of theperiodgene

The clock gene period (per), originally identified inDrosophila, evolves relatively quickly within the insects; probably for this reason, no convincingperhomologues have been identified outside this class. However antibodies toDrosophilaPER have labelled neural pacemakers in other organisms, including mammals. Conserved regions, such as the PAS dimerization domain, reflect its functional importance, but the long Thr–Gly repeat encoded withinperis not conserved outside theDrosophila. The repeat appears to be a component of the temperature compensation system in the fly. This is reflected in the population structure of natural Thr–Gly length variants ofD. melanogaster. Patterns of nucleotide variation withinDrosophila perhave been used to examine the selective forces that have shaped the evolution of this gene.     

Internal rhythms in humans

Human physiology and behavior are characterized by a daily internal temporal dimension. This so-called circadian rhythmicity is present for almost all variables studied to date, persists in the absence of external cycles, and is synchronized to the external 24-h world by an internally generated circadian rhythm of light sensitivity. The light-sensitive circadian pacemaker, presumably also in humans located in the suprachiasmatic nucleus of the hypothalamus, drives the endogenous circadian component of rhythmicity for a number of variables including plasma melatonin, alertness, sleep propensity and sleep structure. Overt rhythmicity and the consolidation of vigilance states are generated by a fine-tuned interaction of this circadian process with other regulatory processes such as sleep homeostasis.     

Oscillations of apoplasmic K+ and H+ activities in Desmodium motorium (Houtt.) Merril. pulvini in relation to the membrane potential of motor cells and leaflet movements

The lateral leaflets of Desmodium motorium exhibit rhythmic upward and downward movements with a period in the minute range. Apoplasmic K⁺ and H⁺ activities were monitored in situ in the abaxial part of the pulvini with ion-selective microelectrodes. An extracellular electric potential was recorded simultaneously. The apoplasmic H⁺ activity of all pulvini exhibiting a regular rhythm of the extracellular electric potential oscillated with the same period between about 10 and 20 mM. The apoplasmic K⁺ activity was high when the membrane potential of the motor cells was depolarized (about 36 mV) and the cells were shrunken. In contrast, the apoplasmic K⁺ activity was low in the swollen state of the motor cells, when the membrane potential was hyperpolarized (about -136 mV). The volatile anesthetic enflurane suppressed reversibly the movement of the leaflets. The same treatment also arrested spontaneous oscillations in the apoplasmic K⁺ activity in the pulvinus. The apoplasmic K⁺ activity oscillated roughly in phase with the K⁺ activity between pH 6.6 and 6.0. Application of white light disturbed the rhythm and increased the extracellular pH. Our results indicate that the physiological mechanism that drives the lateral leaflet movements of Desmodium motorium is closely related to the osmotic motors mediating the leaf movements of Mimosa, Samanea and Phaseolus.Abbreviations E_m membrane potential - E_ex extracellular electric potential - H_ex extracellular H⁺ activity - K_ex extracellular K⁺ activity - R_ex extracellular electrical resistance B. Antkowiak was supported by the Stiftung Volkswagenwerk.     

The role of temperature in the regulation of the circadian rhythm of CO2 fixation in Bryophyllum fedtschenkoi

Detached leaves of Bryophyllum fedtschenkoi Hamet et Perrier kept in normal air show a single period of net CO₂ fixation on transfer to constant darkness at temperatures in the range 0–25 °C. The duration of this initial fixation period is largely independent of temperature in the range 5–20 °C, but lengthens very markedly at temperatures below 4 °C, and is reduced at temperatures above 25 °C. The onset of net fixation of CO₂ on transfer of leaves to constant darkness is immediate at low temperatures, but is delayed as the temperature is increased. The ambient temperature also determines whether or not a circadian rhythm of CO₂ exchange occurs. The rhythm begins to appear at about 20 °C, is most evident at 30 °C and becomes less distinct at 35 °C. The occurrence of a distinct circadian rhythm in CO₂ output at 30° C in the absence of a detectable rhythm in PEPCase kinase activity shows that the kinase rhythm is not a mandatory requirement for the rhythm of PEPCase activity. However, when it occurs, the kinase rhythm undoubtedly amplifies the PEPCase rhythm.Abbreviation PEPCase phosphoenolpyruvate carboxylase We thank the Agricultural and Food Research Council for financial support for this work.     

SPONTANEOUS AND STIMULATED BIOLUMINESCENCE OF THE DINOFLAGELLATE CERATOCORYS HORRZDA (PERIDINIALES)1

This is the first report of spontaneous bioluminescence in the autotrophic dinoflagellate Ceratocorys horrida von Stein. Bioluminescence was measured, using an automated data acquisition system, in a strain of cultured cells isolated from the Sargasso Sea. Ceratocorys horrida is only the second dinoflagellate species to exhibit rhythmicity in the rate of spontaneous flashing, flash quantum flux (intensity), and level of spontaneous glowing. The rate of spontaneous flashing was maximal during hours 2–4 of the dark phase [i.e. circadian time (CT)16–18 for a 14:10 h LD cycle (LD14:10)], with approximately 2% of the population flashing-min^?1, a rate approximately one order of magnitude greater than that of the dinoflagellate Gonyaulax polyedra. Flash quantum flux was also maximal during this period. Spontaneous flashes were 134 ms in duration with a maximum flux (intensity) of 3.1×10⁹ quanta-s^?1. Light emission presumably originated from blue fluorescent microsources distributed in the cell periphery and not from the spines. Values of both spontaneous flash rate and maximum flux were independent of cell concentration. Isolated cells also produced spontaneous flashes. Spontaneous glowing was dim except for a peak of 6.4× 10⁴quanta-s^?1 cell^?1, which occurred at CT22.9 for LD14:10 and at CT22.8 for LD12:12. The total integrated emission of spontaneous flashing and glowing during the dark phase was 4×10⁹ quantacell^?1, equivalent to the total stimulable luminescence. The rhythms for C. horrida flash and glow behavior were similar to those of Gonyaulax polyedra, although flash rate and quantum flux were greater. Spontaneous bioluminescence in C. horrida may be a circadian rhythm because it persisted for at least three cycles in constant dark conditions. This is also the first detailed study of the stimulated bioluminescence of C. horrida, which also displayed a diurnal rhythm. Cultures exhibited >200 times more mechanically stimulated bioluminescence during the dark phase than during the light phase. Mechanical stimulation during the dark phase resulted in 6.7 flashes. cell^?1; flashes were brighter and longer in duration than spontaneous flashes. Cruise-collected cells exhibited variability in quantum flux with few differences in flash kinetics. The role of dinoflagellate spontaneous bioluminescence in the dynamics of near-surface oceanic communities is unknown, but it may be an important source of natural in situ bioluminescence.     

Effects of UV-B radiation and nitrogen starvation on enzyme activities in isolated plasma membranes of Euglena gracilis

Circadian rhythms are characteristic of many physiological and biochemical processes in the freshwater flagellate Euglena gracilis. Earlier, we found that the rhythms of photosynthesis, phototaxis and cell shape followed the same pattern in control organisms, but were differently affected by stress such as UV-B irradiation and nitrogen deficiency. Here we extend our studies to use isolated plasma membranes to characterize the rhythms of some plasma membrane-bound enzymes. Also, we wanted to see whether stress-induced changes of these rhythms could be detected at the subcellular level and possibly be coupled to the changes seen in photosynthesis, phototaxis and cell shape. The isolation of plasma membranes using aqueous polymer two-phase partitioning was successful, as judged by the large enrichment of the plasma membrane-marker 5′-nucleotidase, and the difference in the polypeptide pattern compared with the microsomal fraction from which it was prepared. Two other enzymes were analyzed, K⁺, Mg²⁺-ATPase, and adenylyl cyclase. The specific activities of all three enzymes were decreased by UV-B radiation by ca 30–50%, compared with the control cultures. On the other hand, nitrogen deficiency not only reduced the activity of the K⁺.Mg²⁺-ATPase but also increased the activities of the 5′-nucleotidase and adenylyl cyclase. The different treatments also resulted in differences in polypeptide pattern, e.g., a polypeptide around 30 kDa seemed to be specific to plasma membranes of nitrogen-deficient cultures and one at 39 kDa for the UV-B radiated ones. All three enzymes showed diurnal rhythms that were affected by UV-B radiation. The peak in the rhythm of the ATPase was shifted by UV-B radiation, the rhythm of the 5′-nucleotidase nearly eliminated. The first peak of adenylyl cyclase activity was delayed, so that it looked more like a broad peak between 2 and 11 h after the onset of light. The rhythm of ATPase activity could be correlated with that of photosynthesis in both control and UV-B irradiated cultures. Also, the rhythms of adenylyl cyclase activity and cell shape changes showed some similarities.     

Circadian and seasonal variation of the body temperature of sheep in a tropical environment

Nychthemeral and annual rhythms of the rectal temperature were determined for Corriedale sheep in a tropical climate. The minimum rectal temperature averaged 39.55° C at 0500 hours in summer, and 38.87° C at 0600 hours in winter. The maximum was 40.03° C in summer (1700 hours) and 39.33° C in winter (1830 hours). Annual cycle of the rectal temperature showed a minimum in July and maximum in December.     

An acoustic telemetry study of seabream (Sparus aurata L.): first results on activity rhythm,effects of environmental variables and space utilization

The movements of seabream (Sparus aurata L) were recorded in two earthen ponds (250 m² and 400 m²) by tracking fish tagged with miniature acoustic transmitters. Five seabreams, 500 g in weight, were tracked for periods of four or five days. Fish positions were recorded continuously using an acoustic telemetry system. Each of the five tagged seabreams were tracked individually. Two fishes were released schooled with numerous other individuals, two others were released in isolation and one was grouped with three other individuals.The schooled seabreams were more active in general and, in the large school they were diurnal; isolated fishes however, were more active at night. Temperature influences significantly and progressively the fish activity. Activity decreases at night and increases during daylight. Oxygen saturation also influences swimming activity with a general positive relationship.The horizontal distribution of this species was not uniform. Its resting area was generally located around some particular landmarks such as inlet or outlet pipes in the ponds. No feverish feeding competition was observed between individuals of the same school. The feeding area was cleaned rapidly, in less than 10 minutes, without subsequent returns from the resting area to the feeding area.

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Résumé Les déplacements de la daurade (Sparus aurata L.) ont été enregistrés dans deux bassins en terre (250 m² et 400 m²) en suivant des poissons marqués avec des émetteurs acoustiques miniaturisés. Cinq daurades d'un poids individuel de 500 g furent suivies durant des périodes de 4 ou 5 jours. Les positions des poissons ont été enregistrées en continu avec un systéme de télémétrie acoustique. Chacun des cinq poissons marqués fut suivi individuellement. Deux poissons furent regroupés avec de nombreux autres individus, deux autres furent laissés seuls et le dernier fut mis avec trois autres individus.Les daurades groupées étaient en général plus actives et, en grand banc, étaient diurnes; les poissons isolés étaient toutefois plus actifs durant la nuit. La température influence significativement et progressivement l'activité des poissons. L'activité décroît durant la nuit et augmente pendant le jour. Le pourcentage de saturation en oxygéne agit aussi, de façon positive, sur l'activité de nage.La distribution horizontale de cette espéce n'est pas uniforme. Son aire de repos est généralement localisée dans quelques zones particuliéres comme les arrivées et les sorties d'eau. Aucune compétition alimentaire fébrile n'a été observée entre les individus d'un même banc. L'aire de nourrissage est rapidement nettoyée, en moins de 10 minutes, sans retours ultérieurs depuis la zone de repos jusqu' à la zone de nourrissage.

     

Effects of light-dark cycles on the circadian conidiation rhythm inNeurospora crassa

The effects of 24 hr light-dark cycles on the circadian conidiation rhythm inNeurospora crassa were compared among will-typefrq ⁺ and clock mutantsfrq ⁺,frq ³,frq ⁷,frq ⁹ andfrq ¹¹. The minimum length of the light period necessary for complete entrainment to the light-dark cycles was almost 2 hr infrq ⁺,frq ³ andfrq ⁷ strains. The minimum duration of the dark period necessary for the appearance of circadian conidiation was almost 4 hr in all of the strains except thefrq ¹¹ strain. The phase of the conidiation rhythm was dependent on the light to dark transition in thefrq ¹ strain in all light-dark cycles examined and in thefrq ⁺ andfrq ³ strains when the light period was shorter than 16 hr. In contrast, the phase of thefrq ⁷ strain was dependent on the light to dark transition when the light period was shorter than 10 hr.     

Variable thermal sensitivity as output of a circadian clock controlling the bimodal rhythm of temperature choice in the ant Camponotus mus

Along a thermal gradient and under a LD 1212 h cycle, nurse workers of the ant Camponotus mus select for the brood two different temperatures daily: 30.8°C at the middle of the light period (circadian phase = 90°), and 27.5°C 8 h later, during the dark period (CP = 210°). Brood-carrying activity proved to be self-sustained, running its two daily bursts free with a similar period of 23.5 h, under both LL and DD. The LD alternation acted as a strong Zeitgeber. A phase-delay of the LD 1212 h cycle reset the overt rhythm at once, being both daily events locked-on to the delayed light: dark transition. However, changes in expression, non-occurrence, or even splitting of the two daily brood-carrying events during resetting depended on the phase of the delayed DL transition. By comparing the occurrence of activity with predictions based on a threshold curve of thermal sensitivity, results indicated that an immediate resetting of the involved pacemaker actually takes place. Nurse workers do not directly control the total time spent by the brood at the selected temperature. Instead, the endogenously-driven thermal sensitivity triggers their thermal-searching behavior at two critical times of the day, when environmental temperature is expected to reach its maximum and minimum.