Synchronization of circannual cycles: a cold spring delays the cycles of thirteen-lined ground squirrels

Summary Ground squirrels, show circannual cycles with periods normally less than a year when kept under laboratory conditions. The way in which environmental factors synchronize these cycles with the geophysical year under natural conditions is not known. We tested the possibility that cold temperatures can cause long-term phase delays in circannual cycles of thirteen-lined ground squirrels.Two groups of animals were kept in the cold (4 °C) for either 8.5 or 13 months, after which they were returned to the warm (21 °C) and kept there until they had completed at least one additional cycle. A third group was kept in the warm for the entire experiment. Most of the males in the cold room groups became arrested in the spring phase of their cycles while they were in cold. When returned to the warm, these males resumed cycling. When animals showed prolonged spring phases, their cycles were phase-delayed and continued to reflect this delay even after they were returned to the warm. Both body weight cycles and molt cycles were delayed. In contrast to the males in the cold room, none of the females in the cold room groups and none of the warm room animals of either sex showed this response.Our results demonstrate that cold temperatures can phase-delay both body weight and molt cycles and that the spring phase is a critical stage in this effect. We suggest that spring temperatures are largely responsible for the seasonal synchronization of circannual cycles in ground squirrels and are, therefore, a possible Zeitgeber for these cycles.     

Circannual cycles in golden-mantled ground squirrels: phase shift produced by low temperatures

Summary Golden-mantled ground squirrels,Spermophilus lateralis, were kept in a warm room (21 °C) until they had begun to lose weight following attaining a peak of body weight in the autumn. An experimental group was then transferred to a cold room (–3 °C) and kept there for about 9 months before being returned to the warm room. A control group was kept in the warm room throughout. The peaks in body weight of animals that had been exposed to the cold occurred 4.5 months later than those of the control animals. This difference was maintained when the animals were studied for a further year. Reproductive condition in the experimental animals, after they had been returned to the warm room, also occurred several months later than that in control animals. An extended sojourn in the cold therefore results in a phase shift in circannual cycles of this species.I thank Kirsteen Lang for much help and Linda Jones and others at the Environmental Sciences Centre, Kananaskis, for their cooperation. Janice Farintosh, Yasmin Shah and John Ting assisted with the experiments. The work was supported by the Natural Sciences and Engineering Research Council of Canada.     

Circannual cycles in golden-mantled ground squirrels: Experiments with food deprivation and effects of temperature on periodicity

Summary Female golden-mantled ground squirrels,Spermophilus lateralis, kept in a cool room (9.5 °C), were food deprived and their hibernation thereby extended by 11 weeks on average compared to undeprived control animals in the same room. The deprived group was then refed. Despite their prolonged hibernation, peak weights and onset of hibernation in the subsequent season were only 2–4 weeks later than in the undeprived controls and this effect was only marginally significant. However, the periods of circannual cycles of both the deprived and undeprived groups in the cool room were significantly longer than those of another group of female ground squirrels kept in a warm room (21 °C). It is argued that warm temperatures are capable of terminating the winter phase of circannual cycles, namely that of hibernation and declining body weight. If animals are kept continuously in a cool environment this phase ends spontaneously. Even many weeks of food deprivation are only capable of extending this phase by a relatively small amount, if at all.The author thanks Kirsteen Lang for invaluable help at every phase of this work, Linda Jones and others at the Environmental Sciences Centre, Kananaskis, for heir cooperation, and Janice Farintosh, John Ting and Yasmin Shah for collecting data. Support came from the Natural Sciences and Engineering Research Council of Canada.     

Extensive use of torpor in 13-lined ground squirrels in the fall prior to cold exposure

Mammalian hibernation is characterized by profound reductions in body temperature (T _b) and metabolic, heart and respiratory rates. These reductions are characteristic of torpor, which is temporally confined to winter. Hibernators including ground squirrels are heterothermic in winter, cycling between multiday periods of torpor with low T _b and brief periods of rewarming. In contrast, ground squirrels remain homeothermic during summer, like non-hibernating mammals. The transition between the homeothermic and heterothermic phases of the circannual rhythm of hibernation is often overlooked in hibernation studies. Here, we examined the use of torpor throughout the fall transition in laboratory-housed 13-lined ground squirrels by recording core body temperature with an implanted data logger. As is typical of laboratory-based hibernation studies, animals were kept in standard housing prior to being moved into a cold, dark room to simulate natural hibernation conditions. Significantly, the vast majority of both male and female ground squirrels expressed torpor in the fall while still housed conventionally and prior to cold exposure. The expression of torpor was not predicted by body weight or age, rather it appears to be preprogrammed in a time-dependent manner that is independent of, yet enhanced by, environmental cues. The timing and duration of these torpor bouts occurring prior to cold exposure were also remarkably sporadic. Thus, it is not possible to know with certainty which animals are torpor-naive before cold exposure in the absence of continuous measurement of body temperature. We conclude that fall animals encompass variable points in the transition between summer and winter phases of the circannual cycle of hibernation, thereby confounding studies in which they are used as non-hibernating controls. Conversely, these fall transition animals offer unique opportunities to define the molecular changes that accompany and enable hibernation.     

Phase resetting and phase singularity of an insect circannual oscillator

In circadian rhythms, the shape of the phase response curves (PRCs) depends on the strength of the resetting stimulus. Weak stimuli produce Type 1 PRCs with small phase shifts and a continuous transition between phase delays and advances, whereas strong stimuli produce Type 0 PRCs with large phase shifts and a distinct break point at the transition between delays and advances. A stimulus of an intermediate strength applied close to the break point in a Type 0 PRC sometimes produces arrhythmicity. A PRC for the circannual rhythm was obtained in pupation of the varied carpet beetle, Anthrenus verbasci, by superimposing a 4-week long-day pulse (a series of long days for 4 weeks) over constant short days. The shape of this PRC closely resembles that of the Type 0 PRC. The present study shows that the PRC to 2-week long-day pulses was Type 1, and that a 4-week long-day pulse administered close to the PRC’s break point induced arrhythmicity in pupation. It is, therefore, suggested that circadian and circannual oscillators share the same mode in phase resetting to the stimuli.     

Annual cycles of body weight in the Namie's frosted bat, Vespertilio superans superans

In order to elucidate the circannual cycles of fat deposition and depletion in hibernating bats, annual rhythm of body weight was examined in Vespertilio superans superans under a variety of environmental conditions. Under near natural conditions, adult females put on weight rapidly from October to November. The laboratory born subadult females kept in constant darkness at warm temperature gained weight at the same time as the adults. The length of weight cycle was about 10 months in adult females kept in 14 L : 10 D at about 24°C. These results suggest that cycles of fat deposition and loss are controlled by a modifiable endogenous circannual rhythm. Body weight rhythm persisted normally under warmer environmental conditions preventing the bats from hibernation. It is considered that the circannual cycles of fat gain and loss are relatively stable and not dependent on the hibernation cycle. Prolonged gestation period and delayed beginning of lactation by cold environment in summer did not affect the body weight rhythm. Temperature may be a more important environmental stimulus (or Zeitgeber) for the circannual rhythm than light and its effects may vary with phases.     

A phase response curve for circannual rhythm in the varied carpet beetle <Emphasis Type="Italic">Anthrenus verbasci</Emphasis>

We know that entrainment, a stable phase relationship with an environmental cycle, must be established for a biological clock to function properly. Phase response curves (PRCs), which are plots of phase shifts that result as a function of the phase of a stimulus, have been created to examine the mode of entrainment. In circadian rhythms, single-light pulse PRCs have been obtained by giving a light pulse to various phases of a free-running rhythm under continuous darkness. This successfully explains the entrainment to light-dark cycles. Some organisms show circannual rhythms. In some of these, changes in photoperiod entrain the circannual rhythms. However, no single-pulse PRCs have been created. Here we show the PRC to a long-day pulse superimposed for 4 weeks over constant short days in the circannual pupation rhythm in the varied carpet beetle Anthrenus verbasci. Because the shape of that PRC closely resembles that of the Type 0 PRC with large phase shifts in circadian rhythms, we suggest that an oscillator having a common feature in the phase response with the circadian clock, produces a circannual rhythm.     

Resetting of the circannual rhythm of the varied carpet beetle Anthrenus verbasci by low‐temperature pulses

The varied carpet beetle Anthrenus verbasci L. has a circannual pupation rhythm and pupates in the spring in the wild. The change in photoperiod acts as a predominant zeitgeber for this rhythm. However, it is unclear whether the change in ambient temperature acts as a zeitgeber. The present study examines the effects of low‐temperature pulses on this circannual rhythm by exposing larvae kept under constant short‐day conditions (LD 12 : 12 h) at 20 °C to a lower temperature of 15, 10 or 5 °C for 8 or 12 weeks at various phases. Larval development and pupation are suppressed during exposure to low temperature, with this pupation being induced in sufficiently grown larvae within 2 months of a return to 20 °C. These results are attributed to the exogenous suppression and stimulation of pupation, rather than being related to the circannual rhythm (i.e. masking of the circannual rhythm by temperature). Furthermore, long‐term observations demonstrate the existence of phase‐dependent phase shifts of circannual rhythm as a result of low‐temperature pulses. Circannual phase response curves to low temperature are constructed on the basis of the phase shifts obtained. A low‐temperature pulse as a winter signal can reset the circannual rhythm of A. verbasci. It is probable that both temperature and photoperiod play a role in the entrainment of this circannual rhythm to a natural year.     

Circannual distribution of glioblastoma and anaplastic astrocytoma diagnoses in men and women

Background: circannual variations of incidence may influence human tumors. For example, seasonal distribution of breast cancers suggests an influence of circannual hormonal rhythms. Periodicity of brain tumors has not yet been investigated. The aim of this study was to assess the seasonal variation in glioblastomas and anaplastic astrocytomas. Methods: a large dataset of high-grade gliomas (n = 697) was evaluated with the month of diagnosis as key criterion for time series and seasonal analysis. Circannual and ultra-annual cosinor models were calculated. Results: women suffering from glioblastoma show a peak incidence in fall. No suitable circannual or ultra-annual cosinor model months could be fitted to the data. In women with anaplastic astrocytoma, a peak incidence was detected in summer and fall with a nadir in spring. Here, a significant cosinor model with a period of 8.5 months could be calculated. Conclusion: our data strongly suggest seasonality of diagnosis of high-grade gliomas in women. As gliomas express hormonal receptors, an underlying endocrinal or cell proliferation rhythms may be a possible explanation.     

Effect of winter high temperatures on reproduction and circannual rhythms in hibernating ground squirrels

We tested whether prevention of hibernation in ground squirrels by midwinter exposure to high ambient temperatures influenced timing of the spring phase of reproductive maturation and the phase and period of subsequent circannual rhythms of reproduction and body mass. Exposing hibernating adult male Spermophilus lateralis to 30 degrees C for 6 weeks beginning December 4 advanced the timing of testicular recrudescence by 4-5 weeks, compared to controls left at 4 degrees C. Males exposed to 30 degrees C for 6 weeks beginning at the average time of spontaneous end of hibernation (January 15) reached reproductive maturation at a time intermediate to those of controls and of the December 4 experimental group. However, neither the date of the subsequent fall's body mass peak, the date of the next year's reproductive maturation, nor the periods of circannual rhythms of body mass and reproduction differed among groups. Premature interruption of hibernation appears to allow early expression of reproduction, but does not affect the underlying timing mechanism.     

Behavioural Rhythms of Land Snails in the Field

Circadian and circannual behavioural rhythms of Helix are described in the field. Laboratory studies demonstrate the existence of an endogenous component for these rhythms. The effects of the environmental factors on the snail activities are multiple: light/dark and cold/warm cycles synchronise the behavioural rhythms; a masking effect is observed when it rains since the animals are active throughout the 24 h day; high temperature and low relative humidity cause a complete cessation of activities. In addition, social interactions between and within species can modulate the shape of the rhythms driven by the biological clock.     

Bimodal annual reproductive pattern in laboratory-reared marbled crayfish

The marbled crayfish, Procambarus fallax f. virginalis, is a new research model and potent invader of aquatic ecosystems that reproduces by obligatory parthenogenesis. My data show that in captivity, it can reproduce throughout the year. However, when kept at constant 20 °C and natural photoperiod there were two prominent spawning maxima, one before the spring equinox and another one before the fall equinox. If temperature fluctuated between ~15 °C in winter and ~25 °C in summer, the first maximum was shifted beyond the spring equinox and the second maximum was shifted closer towards the fall equinox. These results indicate a bimodal annual rhythm for the reproduction in marbled crayfish that can be modulated by temperature. Comparison with P. fallax suggests that this rhythmicity was inherited from its sexually reproducing mother species. Potential consequences of my laboratory findings for wild populations of marbled crayfish in temperate and tropical regions are discussed. Since marbled crayfish can be kept in captivity for up to four years under a broad range of conditions, they offer the possibility to investigate endogenous circannual rhythms and their entrainment by environmental zeitgebers in decapod crustaceans.     

Seasonal change in irradiance: a zeitgeber for circannual rhythms in ground squirrels.

1. Zeitgebers for circannual rhythms have been elusive. Demonstration that an external factor is a zeitgeber requires proof of a phase-shift that endures for several years. 2. The California ground squirrel (Spermophilus beecheyi) is an ideal subject. Many features of behavior have circannual rhythms of which change in mass is the easiest to measure. The squirrels thrive in captivity for up to 10 years. The squirrels were kept in individual cages in an air conditioned room, fed lab chow ad lib, and weighed twice a month. They were exposed to a 6-month phase shift of (a) length of day (b) seasonal change in temperature, (c) both, (d) seasonal cycle of irradiance. 3. The squirrels maintained circannual rhythms for up to 9 years. Entrainment was evident only by squirrels exposed to seasonal change in irradiance.     

Circadian and circannual variations in growth and cold resistance of warm acclimated rats and their relationship with the geomagnetic activity

In winter, summer, spring and fall, groups of 200 g Sprague Dawley male rats were kept for 14 days at constant Ta 28°C±°C and LD 12:12(07:00–19:00 h). They were fed a laboratory chow (P) or a semi-purified diet (H). They were weighed at 0, 7 and 14 days. Cold resistance was determined by the fall in rectal temperature during 4 hours of restrained exposure to +1°C either in the morning (09:00 – 13:00 h) or in the afternoon (14:00 – 18:00 h). Rats fed the H diet could better tolerate cold exposure in the morning than in the afternoon, whereas the chow-fed rats were more vulnerable and became severely hypothermic both in morning and afternoon. The greater morning resistance provided by the H diet was evident in summer and winter but not in spring or fall. With both diets, cold resistance as well as growth decreased during spring and fall. Frozen storage of the diets and the water for use during other seasons showed that the nutrients per se were not a factor in the observed seasonal cycle. Although humidity in the 28°C room varied between a minimum of 26.5% in April to a maximum of 44.3% in August, it was not a statistically significant factor in the growth cycle. Arguments are presented to rule out relative humidity as a significant factor in the seasonal variation in the degree of cold resistance. A significant correlation was found for growth and cold resistance with geomagnetic activity.NRCC # 17310     

Verschiebung der Blütezeit der Herbstzeitlose

Summary Potted Colchium plants were subjected to a temperature of 4°C±1° for 2 1/2 to 27 weeks between April and November (1959) and then planted in the garden.Some plants flowered during the fall of the same year in spite of the cold treatment; some of them even flowered during the cold treatment itself. In other plants, the fall flowering of that year was suppressed to different degrees depending on the treatment.Plants that had been exposed to at least 13 weeks of low temperature after the middle of July had some of their fall buds arrested in the development and only flowered in the following spring. Any earlier or later cold treatment was without effect on the time of flowering.Thus the normally fall flowering Colchium was forced by experimental means to flower in spring. This effect resembles the spontaneous spring flowering that is sometimes observed in nature and which has been subject to controversial interpretations.None of the plants that flowered in the spring following the experimental year continued this behavior during consecutive years. The fall flowering of the year following on the experimental year was also influenced by the time and duration of the cold treatment. This indicates that the effects of the cold treatment reached into the next generation of bulbs.

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Mit Unterstützung durch die Deutsche Forschungsgemeinschaft.     

Spermatozoan numbers and characteristics of associated organs in male white-tailed deer during the breeding season

Short-term changes in testicular spermatozoa, testes weight, testes length, epididymes weight, epididymal spermatozoa, and body weight of adult and yearling male white-tailed deerOdocoileus virginianus (Zimmermann, 1780) were examined over a 9-week period that included the peak of the breeding season in South Carolina. Mean values for all characteristics were significantly greater in adults compared to yearlings. Values for reproductive characteristics tended to be highest during the last week in October or the first week in November and decreased over the course of the investigation. Body weight of adults decreased from late October through early December, whereas body weight of yearlings fluctuated only slightly throughout the 9-week period. Significant correlations were observed among all reproductive characteristics and body weight was highly correlated with all characteristics except testicular spermatozoa. Values for reproductive characteristics differed significantly between the right and left sides of the reproductive tracts. Results demonstrate the occurrence of short-term changes in reproductive capabilities of white-tailed deer that are relevant for the development of management strategies for this species.     

Effects of cold,short day and melatonin on thermogenesis,body weight and reproductive organs in Alaskan red-backed voles

Summary This study examined whether cold, short day or melatonin causes reproductive regression and stimulates nonshivering thermogenesis in a subarctic rodentClethrionomys rutilus. Red-backed voles born and raised at 23°C and 22 h light per day (LD 22: 2) at Fairbanks, Alaska (65°N) were exposed in one of six groups to: 1) long day (LD 22:2), 23°C, injected daily with melatonin or saline 2 h before lights out, 2) long day, 3°C, injected daily with melatonin or saline, 3) short day (LD 8:16), 23°C or 3°C. Voles were tested for nonshivering thermogenesis (NST) prior to and after 8 wk exposure. Body weight, testes weight and female reproductive tract weight were assessed after 8 wk in long day and 12 wk in short day.NST was not altered by short day or melatonin but cold (3°C) caused an increase in NST which was similar in long day and short day.Body weight of males and females was not affected by short day but was decreased by melatonin.Short day did not alter mean testes weight (about 20% voles regressed) but reduced mean female reproductive tract weight (more than 40% voles regressed). Melatonin reduced testes weight and female reproductive tract weight (more than 50% of voles of both sexes regressed).The results suggest that in northern red-backed voles: 1) the pineal does not mediate seasonal changes in thermogenic capacity, 2) the pineal may mediate reduction of body weight and regression of reproductive organs but, in addition to daylength, other cues or factors may be important, 3) populations may exhibit variability in sensitivity of reproduction to photoperiod which could allow for opportunistic breeding.Abbreviations NST nonshivering thermogenesis - NE norepinephrine - RMR resting metabolic rate     

Photoperiod and temperature as inducers of gamogenesis in a dicyclic population of Scapholeberis armata Herrick (Crustacea: Cladocera: Daphniidae)

The induction of gamogenesis in a dicyclic population of Scapholeberis armata from a small pond at 40° N in Pennsylvania, USA, was studied. Animals were collected at the time of gamogenesis in mid-October (fall) 1988 and early June (spring) 1989). 10 clones were established from isolated females for each season and maintained at low density under standard conditions of 20 °C and 13.5L/10.5D, with Chlamydomonas reinhardtii diluted with pond water to 3 · 105 cells ml^–1 as a food source. For each clone, mothers of pairs of test animals were acclimated to 4 sets of conditions: 20 °C/15.5L (spring), 15 °C/11.5L (fall), 20 °C/11.5L (control), and 15 °C/15.5L. Test animals were observed bidaily or daily for up to 37 days. Occurrence of molts and of parthenogenetic broods or ephippia was recorded. At short photoperiod, ephippial females developed, usually after one or more parthenogenetic broods, among both spring and fall animals. The response was stronger (higher % sexual females) among fall clones, and was enhanced in both groups by 15 °C water. No animals responded to long photoperiod in warm or cool water. It is likely that additional factors, such as prior photoinduction, rapid depletion of food supply, and increased inter-animal contact due to high population density are required for the induction of spring gamogenesis. Differences in the responsiveness of spring and fall clones to inducing conditions suggest some differences in genotypes of spring and fall subpopulations in this pond. It is proposed that variable responses among genotypes to factors inducing gamogenesis may be the basis for the differences and clines in the timing of gamogenesis found among ecologically and latitudinally separated populations.     

Temperature-induced phase shifting of circadian rhythms in cotton seedlings as related to variations in chilling resistance

The relationship between the degree of chilling resistance and phase shifting caused by low-temperature pulses was examined in two circadian rhythms in cotton (Gossypium hirsutum L. cv. Deltapine 50) seedlings grown under light-dark cycles of 1212 h at 33° C. The seedlings showed a circadian rhythm of chilling resistance and of cotyledon movement. A pulse of 19° C for 12 h during the chilling-sensitive phase (light period) caused a phase delay of 6 h, while a similar temperature pulse during the chilling-resistant phase (dark period) did not cause any phase shift. Exposure to 19° C, 85% RH (relative humidity) for 12 h during the dark period induced chilling resistance in the following otherwise chilling-sensitive light period. In this light period a 12-h 19° C pulse did not cause a phase shift of chilling resistance. Pulses of low temperatures (5–19° C) were more effective in causing phase delays in the rhythm of cotyledon movement when given during the chilling-sensitive phase than when given during the chilling-resistant phase. A 12-h pulse of 5° C, 100% RH during the light period caused a phase delay of cotyledon movement of 12 h. However, when that pulse had been preceded by a chill-acclimating exposure to 19° C, 85% RH for 12 h during the dark period the phase delay was shortened to 6 h. The correlation between higher degree of chilling resistance and the prevention or shortening of the phase delay caused by low temperatures indicates that the mechanism that increases chilling resistance directly or indirectly confers greater ability for prevention of phase shifting by low temperatures in circadian rhythms.Abbreviations CT circadian time - LDC light-dark cycle of 24 h - RH relative humidity     

Effects of winter conditions on Daphnia dynamics and genetic diversity in a dimictic temperate reservoir

1. Winter conditions shape plankton dynamics and community composition in temperate regions, but their effect on dynamics and genetic composition of cyclical parthenogens like Daphnia is largely unclear. 2. For 5 years, we studied the dynamics, hatching from resting eggs and genetic structure of a D. galeata × longispina hybrid complex in a dimictic, temperate reservoir. Our main hypothesis was that higher spring densities and an earlier population peak will be observed after warmer winters, with a lower genetic diversity because of a lower contribution of resting eggs to population growth. 3. The study period could clearly be categorised into cold‐winter years (n = 3) and warm‐winter years (n = 2). Daphnia densities at the end of spring overturn were ～10‐fold lower after cold winters than after warm ones, but no pattern emerged concerning the timing and the height of the population peak in early summer. 4. Hatching intensity from resting eggs was higher and contributed up to 8.5% to Daphnia abundance in a cold‐winter year compared to a negligible contribution in a warm‐winter year. Consistent with this finding, new multilocus genotypes (MLGs) adding to the overwintering stock after the end of spring overturn and presumably originating from resting eggs increased genetic diversity and attained high frequencies within the population only after a cold winter. New MLGs were recorded also after warm winters, but they never gained dominance and no shift in genetic diversity was observed. However, genetic diversity was not generally reduced after warm winters. 5. Our results confirm earlier findings that winter conditions have only a limited effect on the main growth phase and the peak of Daphnia during late spring and early summer. However, winter conditions determine the contribution of resting eggs to the population development, which may profoundly alter the genetic composition of the population compared to the previous season.