Influence of light/dark, seasonal and lunar cycles on serum melatonin levels and synaptic bodies number of the pineal gland of the rat

Synaptic bodies (SB) are ultrastructural organelles observed in the pinealocytes of mammals. According to its shape, they have been classified into synaptic ribbons (SR), synaptic spherules (SS), and intermediate synaptic bodies (ISB). They have been related to the melatonin regulation and production mechanisms of the pineal gland. Circadian and circannual fluctuations of both melatonin and SB have been reported. The possibility that other external factors, apart from light-dark or seasonal cycles, might influence pineal function has been suggested. We studied the evolution of the number of SB and serum melatonin levels not only during light-dark and seasonal phases but also during lunar cycles. Forty male wistar rats were used. Experiment was first carried out in winter and repeated identically in spring. Each season, one group of animals was killed during the new-moon days and a second group during the full-moon days: half of both groups in the photophase and the other half in the scotophase. The number of SB was measured at electron microscopic level whereas serum melatonin levels were determined by radioimmunoassay techniques. Main results showed that SR number and serum melatonin levels were higher during scotophases, winter and full-moon days. The SS only showed a light predominance during winter, whereas predominance of the ISB was found only during the scotophases. These results support the influence of the photophasic factors on the SR and ISB variations. In the case of the SS the influence of the lunar cycles is always dependent on the other factors. Finally, the serum level of melatonin is clearly influenced by the photophasic rhythms and the seasonal periods but not by the lunar cycles.     

Biological adaptability under seasonal variation of light/dark cycles

3A substantial amount of experimental models designed to understand rhythms entrainment and the effects of different regimens of light exposure on health have been proposed. However, many of them do not relate to what occurs in real life. Our objective was to evaluate the influence of “seasonal-like” variation in light/dark cycles on biological rhythms. Twenty adult male Wistar rats were assigned to three groups: control (CT), kept in 12:12 light/dark (LD) cycle; long photoperiod/short photoperiod (LP/SP), kept in 16.5:7.5 LD cycle for 18 days (phase A), then 17 days of gradual reductions in light time (phase B), then 18 days of shorter exposure (7.5:16.5 LD cycle, phase C); short photoperiod/long photoperiod (SP/LP) group, with same modifications as the LP/SP group, but in reverse order, starting phase A in 7.5:16.5 LD cycle. Activity and temperature were recorded constantly, and melatonin and cortisol concentrations were measured twice. Activity and temperature acrophases of all groups changed according to light. The correlation between activity and temperature was, overall, significantly lower for SP/LP group compared with LP/SP and CT groups. Regarding melatonin concentration, LP/SP group showed significant positive correlation between phase A and C (p = 0.018). Animals changed temperature and activity according to photoperiod and demonstrated better adaptability in transitioning from long to short photoperiod. Since this model imitates seasonal variation in light in a species that is largely used in behavioral experiments, it reveals promising methods to improve the reliability of experimental models and of further environmental health research.     

Effect of light/dark cycles on wastewater treatments by microalgae

Chlorella kessleri was cultivated in artificial wastewater using diurnal illumination of 12 h light/12 h dark (L/D) cycles. The inoculum density was 10⁵ cells/mL and the irradiance in light cycle was 45 μmol m² s⁻¹ at the culture surface. As a control culture, another set of flasks was cultivated under continuous illumination. Regardless of the illumination scheme, the total organic carbon (TOC) and chemical oxygen demand (COD) was reduced below 20% of the initial concentration within a day. However, cell concentration under the L/D lighting scheme was lower than that under the continuous illuminating scheme. Thus the specific removal rate of organic carbon under L/D cycles was higher than that under continuous illumination. This result suggested thatC. kessleri grew chemoorganotrophically in the dark periods. After 3 days, nitrate was reduced to 136.5 and 154.1 mg NO₃ ⁻-N/L from 168.1 mg NO₃ ⁻-N/L under continuous illumination and under diurnal cycles, respectively. These results indicate nitrate removal efficiency under continuous light was better than that under diurnal cycles. High-density algal cultures using optimized photobioreactors with diurnal cycles will save energy and improve organic carbon sources removal.     

Influence of medium frequency light/dark cycles of equal duration on the photosynthesis and respiration of Chlorella pyrenoidosa

Chlorella pyrenoidosa was grown in three continuous cultures each receiving a different light regime during the light period of a diurnal cycle. Hourly samples taken during the light period were subjected to medium frequency light/dark oscillations of equal duration, ranging from 3 to 240 seconds. The oxygen consumption and production of each sample were measured with an oxygen electrode in a small oxygen chamber. Although the light/dark cycles had little overall influence on photosynthetic activity, the microalgae appeared to adapt to the light regime to which they were subjected. Large differences were found between the maximum chlorophyll-specific production rates (P _infmax ^supB ), the chlorophyll-specific production rates (P^B) and the respiration rates between the cultures and treated subsamples. Respiration rates increased during the light period, whilst P^B either increased, or had a mid light period minimum or maximum. The culture which received an hourly light oscillation during the light period had the highest P _infmax ^supB and lowest respiration rates, and it is suggested that these algae react as in nature, whereas either a sinusoidal or a block light pattern is unnatural. The latter light regime is commonly used in laboratory studies.     

Efficiency of light utilization of Chlamydomonas reinhardtii under medium-duration light/dark cycles

The light regime inside a photobioreactor is characterized by a light gradient with full (sun)light at the light-exposed surface and darkness in the interior of the bioreactor. Consequently, depending on the mixing characteristics, algae will be exposed to certain light/dark cycles. In this study the green alga Chlamydomonas reinhardtii was cultivated under five different light regimes: (1) continuous illumination; (2) a square-wave light/dark cycle with a light fraction (epsilon) of 0.5 and a duration (t(c)) of 6.1 s; (3) epsilon=0.5, t(c)=14.5 s; (4) epsilon=0.5, t(c)=24.3 s and (5) epsilon=0.8, t(c)=15.2 s. The biomass yield on light energy, protein per photons, decreased under light/dark cycles (epsilon=0. 5) in comparison to continuous light (CL), from 0.207 (CL) to 0.117-0.153 g mol(-1) (epsilon=0.5). Concomitantly, the maximal specific photosynthetic activity, oxygen production per protein, decreased from 0.94 (CL) to 0.64-0.66 g g(-1) h(-1) (epsilon=0.5). Also the quantum yield of photochemistry, yield of the conversion of light energy into chemical energy, decreased from 0.47 (CL) to 0. 23 (epsilon=0.5, t(c)=24.3 s). Apparently, C. reinhardtii is not able to maintain a high photosynthetic capacity under medium-duration light/dark cycles and since specific light absorption did not change, light utilization efficiency decreased in comparison to continuous illumination.     

Influence of gonadotropic hormones on the ultrastructure of rat pinealocytes

Summary The influence of gonadotropic hormones on the ultrastructure of rat pinealocytes was studied. Human chorionic gonadotropin (HCG) as well as pregnant mare serum gonadotropin (PMSG) caused a marked activation of pinealocytes. It is characterised by a conspicuous proliferation of the granular endoplasmic reticulum and Golgi apparatus as well as an increase in number of dense core vesicles, mitochondria, dense bodies, subsurface cisternae and vesicles in the terminal buds of pinealocyte processes. The changes after HCG administration were more pronounced than after PMSG.Supported by a grant from the Polish Academy of Sciences, No. 10.4.2.01.5.6     

Scale-up aspects of photobioreactors: effects of mixing-induced light/dark cycles

The green micro-algae Chlamydomonas reinhardtiiand Dunaliella tertiolecta were cultivated undermedium-duration square-wave light/dark cycles with acycle time of 15 s. These cycles were used to simulatethe light regime experienced by micro-algae inexternally-illuminated (sunlight) air-lift loopbioreactors with internal draft tube. Biomass yieldin relation to light energy was determined as gprotein per mol of photons (400–700 nm). Between 600and 1200 mol m^-2 s^-1 the yield at a10/5 s light/dark cycle was equal to the yield atcontinuous illumination. Consequently, provided thatthe liquid circulation time is 15 s, a considerabledark zone seems to be allowed in the interior ofair-lift loop photobioreactors (33% v/v) without lossof light utilization efficiency. However, at a 5/10 slight/dark cycle, corresponding to a 67% v/v darkzone, biomass yield decreased. Furthermore, bothalgae, C. reinhardtii and D. tertiolecta,responded similarly to these cycles with respect tobiomass yield. This was interesting because they werereported to exhibit a different photoacclimationstrategy. Finally, it was demonstrated that D.tertiolecta was much more efficient at low (average)photon flux densities (57–370 mol m^-2s^-1) than at high PFDs (> 600 mol m^-2s^-1) and it was shown that D. tertiolectawas cultivated at a sub-optimal temperature (20 ^°C).     

A simple model of the growth of phytoplankton populations in light/dark cycles

Phytoplankton populations have been shown to be entrained byalternating periods of light and darkness in natural watersas well as in laboratory cultures. A simple model for the growthof such populations, as reflected by cell division, is presentedhere. The model takes as its structural unit the single cell,using Spudich and Sager's transition point hypothesis for thecoupling between received light and cell cycle progression.A stochastic component is also included to account for cell-to-cellvariability. The model predicts that the characteristics ofcell division patterns in populations entrained by photocyclesdepends mainly on the position of the transition point withinthe cell cycle, rather than on the characteristics of the photocyclicregime. The model simulates successfully the major featuresof observed division patterns of several phytoplankton species.In addition, the model can be used to predict division patternsin high frequency photocycles and during transients inducedby shifts in light regime. Under these conditions, the simulatedpatterns are also consistent with the hypothesis of a circadianclock controlled cell cycle, except in the case of free runningtransients. ¹Present address: Station Biologique Roscoff, CNRS, Roscoff29211, France     

Effects of light and prey availability on nocturnal, lunar and seasonal activity of tropical nightjars

Nightjars and their allies represent the only major group of visually hunting aerial insectivores with a crepuscular and/or nocturnal lifestyle. Our purpose was to examine how both light regime and prey abundance in the tropics, where periods of twilight are extremely short, but nightjar diversity is high, affect activity across different temporal scales. We studied two nightjar species in West African bush savannah, standard‐winged nightjars Macrodipteryx longipennis Shaw and long‐tailed nightjars Caprimulgus climacurus Vieillot. We measured biomass of potential prey available using a vehicle mounted trap and found that it was highest at dusk and significantly lower at dawn and during the night. Based on direct observations, both nightjars exhibit the most intense foraging behaviour at dusk, less intense foraging at dawn and least at night, as predicted by both prey abundance and conditions for visual prey detection. Nocturnal foraging was positively correlated with lunar light levels and ceased below about 0.03 mW m^?2. Over the course of a lunar cycle, nocturnal light availability varied markedly, while prey abundance remained constant at dusk and at night was slightly higher at full moon. Both species increased twilight foraging activity during new moon periods, compensating for the shorter nocturnal foraging window at that time. Seasonally, the pattern of nocturnal light availability was similar throughout the year, while prey availability peaked shortly after onset of the wet season and then slowly decreased over the following four months. The courtship and breeding phenology of both species was timed to coincide with the peak in aerial insect abundance, suggesting that prey availability rather than direct abiotic factors act as constraints, at least at the seasonal level. Our findings illustrate the peculiar constraints on visually orienting aerial nocturnal insectivores in general and tropical nightjars in particular and highlight the resulting nocturnal, lunar and seasonal allocation of activities.     

The hamster clock phase-response curve from summerlike light:dark cycles and its role in daily and seasonal timekeeping

We address the subject of entrainment of the hamster clock by the day:night cycle in summer when the sun sets after 6 PM and rises before 6 AM (nights < 12 h). Summer day:night cycles were simulated by 6 light:dark (LD) cycles with D < 12 h (summerlike, SLD) ranging from SLD 12.5 h:11.5 h (D, 6:15 PM-5:45 AM) to 18 h:6 h (D, 9 PM-3 AM). These are the near limiting SLDs for constant PM timing (entrainment) of behavioral estrus and wheel running in hamsters. The onset of estrus was observed every 4 d in the same hamsters as a phase marker of their 24 h clock. On the day before an experimental estrus, preceded and followed by control onsets, a dark period was imposed to cover a putative 6 PM-6 AM light-sensitive period (LSP). This was scanned with a light pulse (and periodic 5 sec bell alarms) lasting 5-240 min. Shifts in onset of estrus on the next day were plotted vs. the end of the light pulse for PM times ("dusk") and its onset for AM times ("dawn"). The resulting phase shifts from the six SLDs were similar, permitting their combination into a single phase-response curve (PRC) of 1605 shifts. This SLD composite PRC rose at 10:15 PM, peaked at 2 AM (81 min advanced shift), fell linearly to 5:55 AM, and then abruptly to normal at 6 AM (no shift). Peak shift was unaffected by light pulse duration or intensity, or hamster age. The SLD composite PRC lacked the 6 PM-9 PM curve of delayed shifts present in reported PRCs from LD 12 h:12 h and DD. However, a two-pulse experiment showed that all light from 6 PM to L-off was needed to block (balance) the advancing action of a 5 min morning light pulse, thereby maintaining entrainment. A working hypothesis to explain daily entrainment and seasonal fertility in the golden hamster is illustrated. A nomenclature for labeling the phases of the hamster clock (circadian time) is proposed.     

Nitrate utilization in barley: relations to nitrate supply and light/dark cycles

Uptake and utilization of nitrate were investigated in Hordeum vulgare L. cvs Mette and Golf in the vegetative stage, 2 and 4 weeks after sowing. The plants were subjected to a light/dark cycle of 16/8 h (18/12°C). Results obtained with the two genotypes were essentially similar. In the light, xylem nitrate transport and shoot nitrate reduction approximately equalled the amount of nitrate absorbed by the root. A drastic decline in translocation to the shoot in darkness was entirely attributable to decreased transpiration since no major changes in xylem nitrate concentration were observed. Darkening caused only a slight decrease in nitrate uptake, while root nitrate reduction was enhanced. Nitrate starvation for 2 days did not significantlly affect dry matter increment, but resulted in a drastic drop in previously accumulated nitrate, indicating that the stored nitrate is accessible and can sustain unrestricted growth. Uptake increased upon re-addition of nitrate and after 8 h it was about twice that of non-starved plants. During recovery, restoration of root nitrate pools and root nitrate reduction took precedence over shoot nitrate accumulation and reduction. Net nitrate uptake and removal of nitrate from the root to the transpiration stream seem to be decisive for the rate of root nitrate reduction.     

The control of meiosis progression in the fungus Coprinus cinereus by light/dark cycles

Meiosis progression in Coprinus cinereus is controlled by light/dark cycles. Light is essential to propel basidia into karyogamy and light intensity determines the timing of meiotic events. The higher the light intensities, the faster the fruiting bodies enter karyogamy. The critical period when light has this influence is between 16 and 6 h before karyogamy. The control is highly stage specific. A 3-h dark period is essential for a Java dikaryon and the Japanese A(mut)B(mut) homokaryon to enter meiotic metaphase; without it the fruit body is permanently arrested at diffused diplotene. This arrest is light intensity-dependent (>20 hlx) and temperature-dependent (e.g., 27 degrees C). The placement of the dark period is very stage specific; it has no effect when placed before karyogamy stage. A dikaryon of London origin is light blind and able to complete meiosis under continuous high light regime. Fruiting bodies arrested under a continuous high light can be rescued by a 3-h dark treatment, but there is always an 8-h lag time to enter meiotic metaphase. It is possible that the dark effect signals cellular processes leading to division events. Cytological studies of arrested fruiting bodies showed that chromosomes are normal in meiotic prophase through pachytene and diplotene, but are unable to undergo chromosome condensation. Genetic crosses between a monokaryon of Java stock J6;5.4 and a monokaryon BL55 or H5 of London stock showed that light-blindness is dominant, and is controlled by a single Mendelian gene.     

Ultrastructural and biochemical responses of photoreceptor pinealocytes to light and dark in vivo and in vitro

The teleost pineal organ contains functional photoreceptors that synapse with pinealofugal neurons. This study examined the effects of environmental lighting on protein content and levels of putative amino acid transmitters, as well as structural components associated with protein synthesis and neurotransmission. Goldfish subjected to continual illumination for 3 days tended to have increased pineal levels of free amino acids and protein compared to dark adapted glands. Similar responses to environmental lighting occurred in cultured glands suggesting a functional relationship to photosensory mechanisms. Morphometric ultrastructural analyses of pineal photoreceptors showed an increased size of nucleoli (especially the fibrillar component), Golgi bodies, and synaptic ribbons when glands were subjected to continuous light both in vivo and in vitro. The good agreement between protein levels and nucleolar morphology indicates a general effect of environmental lighting on photoreceptor protein metabolism, which may be related to photoreceptor outer segment renewal. Parallel changes in levels of certain amino acids (e.g., glutamate) and size of synaptic ribbons is consistent with an hypothesized role of amino acids in photoreceptor neurotransmission.     

Effect of light/dark cycles on expression of nitrate assimilatory genes in maize shoots and roots

The level of nitrate reductase (NR) and nitrite reductase (NiR) varied in both shoot and root tissue from nitrate-fed Zea mays L. grown under a 16-hour light/8-hour dark regime over a 10-day period postgermination, with peak activity occurring in days 5 to 6. To study the effect of different light regimes on NR and NiR enzyme activity and mRNA levels, 6-day-old plants were grown in the presence of continuous KNO₃ (10 millimolar). Both shoot NRA and mRNA varied considerably, peaking 4 to 8 hours into the light period. Upon transferring plants to continuous light, the amplitude of the peaks increased, and the peaks moved closer together. In continuous darkness, no NR mRNA or NR enzyme activity could be detected by 8 hours and 12 hours, respectively. In either a light/dark or continuous light regime, root NRA and mRNA did not vary substantially. However, when plants were placed in continuous darkness, both declined steadily in the roots, although some remained after 48 hours. Although there was no obvious cycling of NiR enzyme activity in shoot tissue, changes in mRNA mimicked those seen for NR mRNA. The expression of NR and NiR genes is affected by the light regime adopted, but light does not have a direct effect on the expression of these genes.     

Correlations of serotonin and its metabolites in individual rat pineal glands over light:dark cycles and after acute light exposure

Profiles of pineal indolealkylamines were estimated by high performance liquid chromatography and were correlated in individual glands of male rats sacrificed over several light:dark cycles and after acute exposure to light at night. A significant and positive correlation of 5HIAA vs 5HT in individual glands over both normal and experimental lighting conditions suggested that oxidative deamination is not a major factor in photic regulation of pineal 5HT levels and that the formation of 5HIAA is dependent on substrate availability. Regression analysis of other indole constituents revealed that there was a positive and significant correlation between 5HT vs N-acetylserotonin, but not between 5HT vs melatonin and N-acetylserotonin vs melatonin in individual glands during the dark phase of a light:dark cycle. We propose that this effect may be related to a pulsatile release of melatonin into the blood stream and is the result of sampling glands at different stages in the storage/release of melatonin.     

Entrainment of 2 subjective nights by daily light:dark:light:dark cycles in 3 rodent species

Recent work with exotic 24-h light:dark:light:dark (LDLD) cycles indicates surprising flexibility in the entrainment patterns of Syrian hamsters. Following exposure to an LDLD cycle, hamsters may adopt a form of rhythm splitting in which markers of subjective night (e.g., activity, melatonin) are expressed in each of the twice daily scotophases. This pattern contrasts markedly with that of conventionally entrained hamsters in which markers of subjective night are expressed once daily in only 1 of the 2 dark periods. The "split" entrainment pattern was examined further here in Syrian and Siberian hamsters and in mice exposed to LDLD 7:5:7:5, a condition that reliably induces split activity rhythms in all 3 species. The phase angle of entrainment and activity duration were generally similar comparing the 2 daily activity bouts in each species. The stability of this split entrainment state was assessed by deletions of photophases on individual days, by exposure to skeleton photoperiods, and by transfer to constant darkness. As in Syrian hamsters, the one-time substitution of darkness for one 7-h photophase did not grossly alter activity patterns of Siberian hamsters but acutely disrupted the split rhythms of mice. Skeleton light pulses of progressively shorter duration did not significantly alter split entrainment patterns of either Syrian or Siberian hamsters. Both species continued to exhibit stable entrainment with activity expressed in alternate scotophases of an LD 1:5 cycle presented 4 times daily. In contrast, the split activity rhythms of mice were not maintained under skeleton pulses. In constant darkness, rhythms of Siberian hamsters remained distinctly split for a minimum of 2 cycles. Split entrainment to these novel LDLD and 4-pulse skeleton lighting regimes demonstrates a marked degree of plasticity common to the circadian systems of several rodent species and identifies novel entrainment patterns that may be reliably elicited with simple environmental manipulations. Inter- and intraspecific differences in the stability of split activity rhythms likely reflect differences in coupling interactions between the component circadian oscillators, which, adopting separate phase relations to these novel LD cycles, yield a split entrainment pattern.     

Chronobiotic effect of melatonin following phase shift of light/dark cycles in the field mouseMus booduga

The objective of this study was to assess whether melatonin accelerates the re-entrainment of locomotor activity after 6 h of advance and delay phase shifts following exposure to LD 12:12 cycle (simulating jet-lag/shift work). An experimental group of adult male field mice Mus booduga were subjected to melatonin (1 mg/kg) through i.p. and the control group were treated with 50 % DMSO. The injections were administered on three consecutive days following 6h of phase advance and delay, at the expected time of “lights off”. The results show that melatonin accelerates the re-entrainment after phase advance (29%) when compared with control mice. In the 6 h phase delay study, the experimental mice (melatonin administered) take more cycles for re-entrainment (51%) than the control. Further, the results suggest that though melatonin may be useful for the treatment of jet-lag caused by eastward flight (phase advance) it may not be useful for westward flight (phase delay) jet-lag