Seasonal expression patterns of clock-associated genes in the blue mussel Mytilus edulis

Environmental cues allow organisms to synchronise their internal biological rhythms with external environmental cycles. These rhythms are regulated on a molecular level by oscillating interactions between clock genes and their proteins. Light is a particularly relevant environmental cue, provisioning daily information via light/dark cycles as well as seasonal information via day-length (photoperiod). Despite the ecological and commercial importance of bivalves, little is known about the interactions comprising their molecular clock mechanism. This study investigates the link between the annual seasonal progression and reproductive development in the blue mussel (Mytilus edulis), using mRNA expression patterns of clock-associated genes: Clock, Cry1¸ ARNT, Timeout-like, ROR/HR3 and aaNAT, in the gonads of both sexes, sampled over three daily time-points on a tidal beach during the winter and summer solstices. Significant differences in mRNA expression levels, including some seasonal differences at comparable time-points, were detected for all genes with the exception of Timeout-like. These differences occurred seasonally within sex (Clock, Cry1, ROR/HR3), seasonally between sexes (Clock, Cry1, ARNT, ROR/HR3, aaNAT) and daily between sexes (Cry1), although no significant daily differences were detected in summer or winter for either sex for any of the genes. This study reveals that clock-associated genes show seasonal responses in this species of bivalve. Understanding the mechanisms by which environmental cues drive biological rhythms is critical to understanding the seasonal sensitivity of this keystone species to environmental changes.     

Light and dark rations and the photic entrainment of circadian locomotor activity patterns in the South American Silver Catfish (Rhamdia quelen,Quoy & Gaimard, 1824)

The aim of this study was to evaluate the daily rhythm of locomotor activity in Rhamdia quelen (R. quelen). A total of 30 fish were enrolled in the study and were equally divided in 10 groups and maintained in 100 liters tanks. The locomotor activity was measured in fish maintained under the LD 12:12 photoperiod regime; thereafter, the LD cycle was reversed to DL in order to study the resynchronization and to explore the endogenous pacemaker. Subsequently, the fish were subjected to constant conditions of light to test whether or not locomotor rhythms are regulated by the endogenous circadian clock. The effect of increasing light length and intensity was studied on daily rhythm of locomotor activity of fish. Our results showed that the R. quelen is a strictly diurnal species, the rhythm of locomotory activity resynchronized quickly after inverting the LD cycle and persist under free course LL, suggesting a circadian origin. The light showed a significant masking effect often blocking the expression of the biological rhythm. The strictly diurnal behavior is controlled directly by the photoperiod and maintained even under very dim light (30 lux).     

Seasonal changes in food consumption and growth of Arctic charr exposed to either simulated natural or a 12:12 LD photoperiod at constant water temperature

Immature male and female and maturing male Arctic charr Salvelinus alpinus were held at constant temperature (4° C) under either simulated natural photoperiod (nLD) or constant photoperiod (12:12 LD) conditions, and feed intake and growth were monitored at monthly intervals over 13 months. Food was supplied in excess during a 5-h period each day, and daily consumption was determined using X-radiography. Food consumption and growth of both immature and maturing fish showed distinct seasonal cycles irrespective of photoperiod treatment. Feed intake and growth rates were highest in mid-summer and lowest in autumn. The fact that seasonal cycles persisted under constant photoperiod (12:12 LD) and temperature conditions suggests that circannual rhythms of food consumption and growth exist in the absence of seasonal changes in these environmental cues. The data support the view that seasonal changes in food consumption and growth in the Arctic charr are driven by endogenous rhythms. Reductions in feeding and growth in autumn occurred about 1 month earlier in the maturing males than in the immature fish. Males that matured under the 12:12 LD regime displayed a growth cycle that was delayed in comparison with that shown by the maturing males held under nLD.     

Circadian rhythms of locomotor activity and temperature selection in sleepy lizards, <Emphasis Type="Italic">Tiliqua rugosa</Emphasis>

This study examined whether the daily rhythms of locomotor activity and behavioural thermoregulation that have previously been observed in Australian sleepy lizards (Tiliqua rugosa) under field conditions are true circadian rhythms that persist in constant darkness (DD) and whether these rhythms show similar characteristics. Lizards held on laboratory thermal gradients in the Australian spring under the prevailing 12-hour light : dark (LD) cycle for 14 days displayed robust daily rhythms of behavioural thermoregulation and locomotor activity. In the 13-day period of DD that followed LD, most lizards exhibited free-running circadian rhythms of locomotor activity and behavioural thermoregulation. The predominant activity pattern displayed in LD was unimodal and this was retained in DD. While mean levels of skin temperature and locomotor activity were found to decrease from LD to DD, activity duration remained unchanged. The present results demonstrate for the first time that this species’ daily rhythm of locomotor activity is an endogenous circadian rhythm. Our results also demonstrate a close correlation between the circadian activity and thermoregulatory rhythms in this species indicating that the two rhythms are controlled by the same master oscillator(s). Future examination of seasonal aspects of these rhythms, may, however, cause this hypothesis to be modified.     

Novel masking effects of light are revealed in Drosophila by skeleton photoperiods

Here, we show that in a skeleton photoperiod where all midday light is removed from a standard laboratory 12:12 LD photoperiod, a large diurnal peak of activity is revealed that is continuous with the E peak seen in constant dark (DD). We further show that the circadian clock gene tim regulates light-dependent masking of daytime activity, but the clock gene per does not. Finally, relative to wild-type flies, mutants for both of these clock genes showed increased nighttime activity in the skeleton photoperiod but not in the standard photoperiod. This result suggests that nighttime activity is suppressed by the intact circadian clock, and in its absence, by exposure to a standard photoperiod. These results support and extend the literature addressing the complex interactions between masking and clock-controlled components of overt circadian rhythms.     

Photoperiod affects the diurnal rhythm of hippocampal neuronal morphology of siberian hamsters

Individuals of many species can regulate their physiology, morphology, and behavior in response to annual changes of day length (photoperiod). In mammals, the photoperiodic signal is mediated by a change in the duration of melatonin, leading to alterations in gene expressions, neuronal circuits, and hormonal secretion. The hippocampus is one of the most plastic structures in the adult brain and hippocampal neuronal morphology displays photoperiod-induced differences. Because the hippocampus is important for emotional and cognitive behaviors, photoperiod-driven remodeling of hippocampal neurons is implicated in seasonal differences of affect, including seasonal affective disorder (SAD) in humans. Because neuronal architecture is also affected by the day-night cycle in several brain areas, we hypothesized that hippocampal neuronal morphology would display a diurnal rhythm and that day length would influence that rhythm. In the present study, we examined diurnal and seasonal differences in hippocampal neuronal morphology, as well as mRNA expression of the neurotrophic factors (i.e., brain-derived neurotrophic factor [Bdnf], tropomyosin receptor kinase B [trkB; a receptor for BDNF], and vascular endothelial growth factor [Vegf]) and a circadian clock gene, Bmal1, in the hippocampus of Siberian hamsters. Diurnal rhythms in total length of dendrites, the number of primary dendrites, dendritic complexity, and distance of the furthest intersection from the cell body were observed only in long-day animals; however, diurnal rhythms in the number of branch points and mean length of segments were observed only in short-day animals. Spine density of dendrites displayed diurnal rhythmicity with different peak times between the CA1 and DG subregions and between long and short days. These results indicate that photoperiod affects daily morphological changes of hippocampal neurons and the daily rhythm of spine density, suggesting the possibility that photoperiod-induced adjustments of hippocampal neuronal dynamics might underlie seasonal difference of affective responses. Bmal1 mRNA showed a diurnal rhythm and different expression levels between long and short days were observed. However, there were no strong effects of day length on Bdnf. trkb, and Vegf gene expression, suggesting that these genes are not involved in the photoperiodic effects on hippocampal neurons.     

Coexistence of congeneric spiny lobsters on coral reefs: differential use of shelter resources and vulnerability to predators

The notion, previously generated from laboratory experiments, that the local coexistence of spiny lobsters Panulirus guttatus and Panulirus argus on Caribbean reefs may be promoted by a differential use of shelter resources and/or vulnerability to predators was explored in a coral reef in Mexico. Multiple regressions with data collected on 11 occasions on fixed back-reef and fore-reef sites suggested that the densities of the congener, other crevice-dwellers, and predators did not significantly affect the density of P. argus in either reef zone, or of P. guttatus on the back-reef, where coexistence of both lobster species was greatest. In contrast, there was a significant negative relationship between predators and the density of P. guttatus on the fore-reef, where this species was dominant. Congeneric cohabitation in dens was less than expected by chance, but this pattern may reflect a differential use of shelter resources rather than interspecific competition. P. guttatus was more prevalent in dens over the middle and upper third of the reef profile, and P. argus over the lower and middle third of the reef profile. Whether individuals cohabited with conspecifics, congeners, or resided solitarily, P. guttatus was more prevalent at the walls and/or ceiling and P. argus on the floor of dens. This differential use of shelter resources may be related to the differential vulnerability to predators, which may have promoted local coexistence of these congeners in reef habitats.     

Circadian clock of Drosophila montana is adapted to high variation in summer day lengths and temperatures prevailing at high latitudes

Photoperiodic regulation of the circadian rhythms in insect locomotor activity has been studied in several species, but seasonal entrainment of these rhythms is still poorly understood. We have traced the entrainment of activity rhythm of northern Drosophila montana flies in a climate chamber mimicking the photoperiods and day and night temperatures that the flies encounter in northern Finland during the summer. The experiment was started by transferring freshly emerged females into the chamber in early and late summer conditions to obtain both non-diapausing and diapausing females for the studies. The locomotor activity of the females and daily changes in the expression levels of two core circadian clock genes, timeless and period, in their heads were measured at different times of summer. The study revealed several features in fly rhythmicity that are likely to help the flies to cope with high variation in the day length and temperature typical to northern summers. First, both the non-diapausing and the diapausing females showed evening activity, which decreased towards the short day length as observed in the autumn in nature. Second, timeless and period genes showed concordant daily oscillations and seasonal shifts in their expression level in both types of females. Contrary to Drosophila melanogaster, oscillation profiles of these genes were similar to each other in all conditions, including the extremely long days in early summer and the cool temperatures in late summer, and their peak expression levels were not locked to lights-off transition in any photoperiod. Third, the diapausing females were less active than the non-diapausing ones, in spite of their younger age. Overall, the study showed that D. montana clock functions well under long day conditions, and that both the photoperiod and the daily temperature cycles are important zeitgebers for seasonal changes in the circadian rhythm of this species.     

Seasonal pre‐migratory fattening and increased activity in a nomadic and irruptive migrant,the Red Crossbill Loxia curvirostra

It is well documented that irruptive and nomadic migrants move in response to resources that are distributed unpredictably in space, time or both. Increasing evidence, however, suggests that irruptive and nomadic species may use seasonal timing mechanisms to prepare for migrations, despite the more facultative nature of their movements. Here we use data from free‐living and captive Red Crossbills Loxia curvirostra, a typical irruptive nomad, to examine three hypotheses regarding control of facultative migration: (1) the facultative migration hypothesis, which states that both preparation and departure decisions are regulated by resource availability; (2) the seasonal preparation hypothesis, which states that preparation is initiated by seasonal factors (i.e. endogenous rhythms and/or photoperiod) but that departure decisions are dependent on local resource availability; and (3) the seasonal migration hypothesis, which states that both preparation and departure decisions are initiated by seasonal mechanisms and are independent of local food resources. Red Crossbills in North America are thought to make temporally consistent spring migrations in anticipation of conifer cone maturation. In this study, fat deposits of free‐living Red Crossbills peaked in May and June, exceeding even winter deposits. In agreement with the field data, captive birds on a natural photoperiod with constant food and temperature showed a peak in fat deposition and activity levels in June. These findings are consistent with the seasonal preparation and the seasonal migration hypotheses and contribute to a growing literature that suggests that facultative migrants may prepare for movements in similar ways to seasonal migrants.     

Daily and annual variations of the hepatic glucose 6-phosphate dehydrogenase activity and seasonal changes in the body fats of the gilthead seabream Sparus aurata

In mammalians, Glucose 6-phosphate dehydrogenase (G6PDH) is clearly related with the daily cycle lipogenesis/lipolysis and therefore with the photoperiod and activity. In fish the temporally changes on the activity of this enzyme have not been described. In this work, annual and diurnal activity of G6PDH was investigated in the liver of Sparus aurata in different seasonal conditions of temperature and photoperiod. During experiment the fish were kept in cages under natural temperature and photoperiod while consuming a commercial diet. The months studied were chosen to establish if there is any influence on the G6PDH activity, because of the change in temperature and photoperiod (equal photoperiod and different temperature, March and October; different photoperiod and equal temperature, May and November; and different photoperiod and temperature, June and January). To study the enzyme activity along the day, the livers of six fish were removed every 3 hr during 24 hr. Annual activity was determined as the average activity during 1 day. Also the annual changes on fat content in muscle and digestive system were studied. The Vmax (maximum velocity) showed seasonal changes in period and phase of the rhythms and for the Km (Michaelis-Menten constant) a period of 24 hr was found in all sampled months, except March, the phase time moves depending on the month. In addition, the annual variations in muscular fat and G6PDH-specific activity seem to be more related to the reproductive stage than any other variable considered.     

Loss of the circadian rhythms of locomotor activity, food intake, and plasma melatonin concentration induced by constant bright light in the pigeon (Columba livia)

Summary Locomotor activity and feeding activity were measured together with circulating levels of melatonin in pigeons which were exposed to constant bright light (LLbright, 2000 lux) following light-dark (LD) cycles. Although all the pigeons showed daily rhythms of locomotor activity, feeding activity, and melatonin levels under LD cycles, they lost all the rhythms in prolonged LLbright. Acute exposure to bright light (2000 lux) during darkness reduced plasma melatonin levels. The half-time for the suppression in melatonin levels was about 30 min after short-term light exposure. These results support the hypothesis that melatonin may control the circadian rhythms of locomotor activity and feeding activity in the pigeon.Abbreviations LD light-dark - LLdim constant dim light - LLbright constant bright light - DD constant darkness - PX pinealectomy - EX blinding - RIA radioimmunoassay     

Pineal and retinal melatonin is involved in the control of circadian locomotor activity and body temperature rhythms in the pigeon

Summary Although pinealectomy or blinding resulted in loss of the clarity of the free-running rhythm of locomotor activity and body temperature and reduced the peak level of circulating melatonin rhythms to approximately a half in intact pigeons, neither pinealectomy nor blinding abolished any of these rhythms. However, when pinealectomy and blinding were combined, the rhythms of locomotor activity and body temperature disappeared in prolonged constant dim light, and melatonin concentration was reduced to the minimum level of detection. In order to examine the role of melatonin in the pigeon's circadian system, it was administered either daily or continuously to PX + EX-pigeons in LLdim. Daily administration of melatonin restored circadian rhythms of locomotor activity which entrained to melatonin injections, but continuous administration did not induce any remarkable change of locomotor activity. These results suggest that melatonin synthesized in the pineal body and the eye contributes to circulating melatonin and its rhythmicity is important for the control of circadian rhythms of locomotor activity and body temperature in the pigeon.Abbreviations LD Light-dark - LLdim constant dim light - LLbright constant bright light - PX pinealectomy - EX blinding - SCN suprachiasmatic nucleus     

Crepuscular activity of adult Sericesthis geminata (Coleoptera: Scarabaeidae): Influence of circadian rhythmicity and light intensity

Subterranean Sericesthis geminata (Boisduval) beetles emerge from the soil daily during the flight season. Emergence and flight are initiated within 15 min after sunset. The beetles are active above the soil surface as the intensity of illumination decreases from 240 lux to 0.7 lux.

If young adult beetles still in their pupal cells in the soil are placed under constant, low‐intensity illumination, they first emerge at any time of day, showing no cyclical diurnal activity pattern. When such beetles are exposed to diurnal fluctuations in illuminance, the first emergence and flight occur at dusk. These beetles subsequently show a diurnal rhythm in their activity, even when exposed to constant environmental conditions. This endogenous activity cycle has a period of about 22 h, and is synchronised with diurnal fluctuations in light intensity. It is reset in response to a change in the photoperiod.

The data suggest that, after initial emergence from the soil, daily crepuscular activity is initiated by an endogenously controlled activity rhythm which causes the beetles to burrow to the soil surface shortly before dusk. At the soil surface they are exposed to fluctuations in the duration and intensity of illumination which may reset the endogenous rhythm, affecting the time of subsequent emergence.     

Biophysical connectivity explains population genetic structure in a highly dispersive marine species

Connectivity, the exchange of individuals among locations, is a fundamental ecological process that explains how otherwise disparate populations interact. For most marine organisms, dispersal occurs primarily during a pelagic larval phase that connects populations. We paired population structure from comprehensive genetic sampling and biophysical larval transport modeling to describe how spiny lobster (Panulirus argus) population differentiation is related to biological oceanography. A total of 581 lobsters were genotyped with 11 microsatellites from ten locations around the greater Caribbean. The overall F _ST of 0.0016 (P = 0.005) suggested low yet significant levels of structuring among sites. An isolation by geographic distance model did not explain spatial patterns of genetic differentiation in P. argus (P = 0.19; Mantel r = 0.18), whereas a biophysical connectivity model provided a significant explanation of population differentiation (P = 0.04; Mantel r = 0.47). Thus, even for a widely dispersing species, dispersal occurs over a continuum where basin-wide larval retention creates genetic structure. Our study provides a framework for future explorations of wide-scale larval dispersal and marine connectivity by integrating empirical genetic research and probabilistic modeling.

     

Photoperiod and cell mediated immunity of clinically normal dogs

Abstract

Previously, the authors have reported seasonal variations in cell mediated immunity in the dog during the period July, 1977 ‐ October 1978 as measured by whole blood lectin‐induced lymphocyte transformation. Peak activity occurred in the summer, suggesting association with photoperiodicity. Here the authors report on immune response of dogs kept indoors ‐ under controlled physical environment ‐ with a natural (outdoor) photoperiod or under a 12:12 h (LD) regime, and a control group kept in outdoor kennels. Peak immune activity in 1979 occurred in the winter, in both indoor groups as well as the outdoor groups subject to natural photoperiod. Since the indoor dogs were kept at a constant temperature and humidity in clean (filtered) air, photoperiod, temperature, and particulate air contaminants probably are not associated with seasonal variations in immunity. The underlying cause for either the seasonal variations or the shift from peak activity in the summer of 1978 to winter of 1979 is unknown. Dogs under LD = 12:12 light regime had a significantly lowered immunity relative to the dogs with the natural photoperiod.     

How photoperiod influences body temperature selection in Lacerta viridis

Summary European green lizards, Lacerta viridis, show a distinct annual cycle in their day and nighttime selected body temperature (T _b) levels when monitored under natural photoperiod. The amplitude between daily photophase and scotophase temperatures varies throughout the year. Highest body temperatures with smallest day/night variation are selected from May through July. Throughout fall, the difference between day and nighttime selected T _b levels increases. Lizards inevitably enter a state of winter dormancy which terminates daily rhythmicity patterns. Under natural photoperiodic conditions, cessation of dormancy occurs spontaneously by mid-March, regardless whether high temperatures are available or not. Lacerta viridis respond to an artificial long photoperiod (16 h light, 8 h dark) at all times of the year with modifications in both diel patterns and levels of selected T _b to summer-like conditions. When, however, the natural photoperiod at different phases in the annual cycle is held constant for six to eight weeks, T _b selection of Lacerta viridis also remains stable at the level corresponding to the prevailing photoperiod. These results implicate that the photoperiod is a more prominent Zeitgeber for seasonal cueing of temperature selection than has been surmised in the past. Further, we suggest that the large variations recorded in daily T _b cycles do not imply that this lizard is an imprecise thermoregulator, but rather indicates an important integral process necessary for seasonal acclimatization.     

The sensory innervation of the foregut of Panulirus argus (Decapoda Crustacea)† ‡

Ths structure of the stomatogastric neuromuscular system in Panulirus argus, Callinectes sapidus and Homarus americanus has recently been described (Maynard and Dando, 1973). We attempt here to describe the sensory innervation of the foregut in Panulirus argus and, by combining this information with previous published data and less systematic observations on Callinectes and Homarus, to provide in addition a summary of the stomatogastric sensory systems in these types of Decapoda Crustacea (Figure 1, Table I).

Some anatomical problems remain unresolved and there is variation in the structure of the sense organs in different species, but we are able to recognize six major receptor groups in all of the species examined. These are (i) mechanoreceptors which monitor movements of the lower oesophagus and mouth (Receptor reference Nos. 1, 2, 3); (ii) probable chemoreceptors in the higher oesophagus and ventral cardiac sac (Rf. Nos. 6, 9, 11); (iii) cells located in or near the stomatogastric ganglion which monitor movements of the gastric mill (Rf. No. 8); (iv) neurones of the posterior stomach nerve (Rf. No. 15) which monitor movements of the gastric mill; (v) neurones innervating muscles near the cardio‐pyloric valve (Rf. No. 16); (vi) neurones innervating the hepatopancreas duct and the initial part of the intestine (Rf. Nos. 18, 19).

In such a restricted system it should be possible to determine the precise role that the various sensory systems play in the control of the simple movements of the foregut. This research must necessarily involve the investigation, with intracellular techniques, of the central events in the commissural ganglia as these ganglia appear to be the major co‐ordination centres of the stomatogastric nervous system.     

Daily rhythms of clock gene expression and feeding behavior during the larval development in gilthead seabream,Sparus aurata

Light is the main environmental time cue which synchronizes daily rhythms and the molecular clock of vertebrates. Indeed, alterations in photoperiod have profound physiological effects in fish (e.g. reproduction and early development). In order to identify the changes in clock genes expression in gilthead seabream larvae during ontogeny, three different photoperiods were tested: a regular 12L:12D cycle (LD), a continuous light 24L:0D (LL) and a two-phases photoperiod (LL?+?LD) in which the photoperiod changed from LL to LD on day 15 after hatching (dph). Larvae were sampled on 10, 18, 30 and 60 days post-hatch (dph) during a 24?h cycle. In addition to the expression of clock genes (clock, bmal1, cry1 and per3), food intake was measured. Under LD photoperiod, larvae feed intake and clock genes expression showed a rhythmic pattern with a strong light synchronization, with the acrophases occurring at the same hour in all tested ages. Under LL photoperiod, the larvae also showed a rhythmic pattern but the acrophases occurred at different times depending on the age, although at the end of the experiment (60 dph) clock genes expression and feed intake rhythms were similar to those larvae exposed to LD photoperiod. Moreover, the expression levels of bmal1 and cry1 were much lower than in LD photoperiod. Under the LL?+?LD photoperiod, the 10 dph larvae showed the same patterns as LL treatment while 18 and 30 dph larvae showed the same patterns as LD treatment. These results revealed the presence of internal factors driving rhythmic physiological responses during larvae development under constant environmental conditions. The LL?+?LD treatment demonstrates the plasticity of the clock genes expression and the strong effect of light as synchronizer in developing fish larvae.     

Behavioral rhythms of the Japanese newts, Cynops pyrrhogaster, under a semi-natural condition

 Locomotor activity rhythms of the Japanese newt, Cynops pyrrhogaster, were recorded under a semi-natural condition using phototransistor systems. The daily activity rhythm showed a seasonal change: the locomotor activity was mainly diurnal (active during the daytime) from spring to early summer; mainly nocturnal (active during the night-time) from summer to autumn; and showed either a diurnal or nocturnal pattern, depending on the ambient temperature, in winter. To analyze the daily activity in detail, we observed the behavior of a group of newts (three males, three females) throughout 24 h. Four types of behavior (respiration, feeding, mating, and resting on the land) were observed. Each behavior had daily rhythms and showed a seasonal change. The behavior on land showed mainly a nocturnal or bimodal pattern (activity rhythms with two peaks) throughout the year and was more frequently observed in summer. Mating behavior also showed a seasonal change: high activity in spring, with peaks in the early morning and evening, but no activity in summer. Except in winter, feeding and respiratory behavior showed no seasonal changes in either activity period or frequency. Coupling between behavior and the clock seems to be weak in the Japanese newt because of indistinct daily rhythms and frequent phase changes of locomotor activity in water. Physical factors such as humidity and temperature seem to affect strongly the daily activity of the newts. Received: 21 April 1997 / Accepted: 1 September 1997