Daily variations in the sensitivity of soybean seedlings to low temperature

Every 4 hr during a time span of 32 hr and of 76 hr a different group of soybean seedlings [Glycine max (L.) Merr., cv. Corsoy] maintained under a regime of 16 hr of light followed by 8 hr of darkness was exposed to -10 degrees C for 4 min. Extent of sensitivity to low temperature was evaluated approximately 10 days after exposure to freezing temperature by determining the weight of the plants and chlorophyll content of the cotyledons. The amount of sensitivity to low temperature was related to the time of exposure and displayed a significant 24-hr oscillation. Plants appeared to be least sensitive to cold injury during the late portions of the light span. Plants subjected to water stress were less sensitive to cold and no significant 24-hr oscillation in response to low temperature could be detected by the cosinor method of analysis.     

Environmental factors controlling the time of flower-opening in Pharbitis nil

Pharbitis nil, strain Violet, subjected to various photoperiods(24-hr cycle at 24?C) bloomed about 10 hr after light-off whenthe light period was 10 hr or longer, and about 20 hr afterlight-on when the light period was shorter. The higher the temperature(20–30?C) during the dark period, the later the time offlower-opening, with the temperature during the last half ofthe dark period having a stronger effect than that during thefirst half. In continuous dark or light, flower buds of Pharbitis openedabout every 24 hr at all temperatures tested between 20 and28?C, which suggests the participation of a circadian rhythmin determining the time of flower-opening. A light pulse given6–12 or 28–36 hr after the onset of the dark periodgreatly advanced the phase of this rhythm (8–10 hr). Phasedelay of this rhythm could not be obtained by light pulses givenat any time. (Received September 29, 1979; )     

Sequential study on the influence of adriamycin on cell proliferation and ultrastructure of cultured mammary tumor cells

The time-dependent cytocidal and growth inhibitory effects of Adriamycin (ADM) on monolayer cultures of 7,12-dimethylbenz[a]anthracene (DMBA)-induced mammary tumor cells were analyzed. The inhibitory effect on cell proliferation was assessed by colony formation in soft agar. Growth inhibition and [3H]thymidine labeling indices clearly demonstrate a dose-dependent antimitotic and cytotoxic effect of the drug. At low concentrations (10(-9)-10(-8) M), 90-100% of cells survived 24-hr exposure. At a higher concentration (10(-5) M), 75-80% of cells survived after 8-hr exposure; by 72 hr only 20-30% of the cells remained. Autoradiographic examination of the pulse-labeled cultures demonstrated no change in the proportion of cells in S-phase during the first 4 hr of treatment. Subsequently DNA synthesis was completely abolished and remained inhibited for the duration of the experiment (72 hr). Clonogenic assay revealed a complete arrest of growth in cells exposed to 10(-5) M ADM and greater than 60% inhibition of cell proliferation at 10(-7) M. Ultrastructural changes were not observed in cells during the first 4 hr of treatment; however, after 8 hr most surviving cells exhibited alterations in nuclear chromatin. The surviving cells showed mitochondrial degeneration, myelin body formation, and vacuolization of the endoplasmic reticulum. This study shows the potential usefulness of the primary culture system in drug evaluation. In addition, serial observation of the effects of ADM revealed a cell subpopulation of the primary culture with differential sensitivity to the drug.     

Mineralization in rat metaphyseal bone exhibits a circadian stage dependency

Density gradient fractionation analysis of rat metaphyseal bone was used to delineate the biorhythmic changes in bone matrix mineralization. Seventy-two 4-week-old rats were entrained to 12-hr light, 12-hr dark cycles (light, 0800-2000 hr; darkness, 2000-0800 hr) for 4 weeks. All animals were fed ad lib. on Purina laboratory rat chow and tap water. Groups of 10-12 rats were killed by cervical dislocation at 4-hr intervals during a 24-hr period, and the tibias were then biopsied and frozen in liquid N2. Metaphyseal bone was fractionated via bromoformtoluene density gradients into specific gravity fractions ranging from 1.7 to 2.8. Density gradient fractions were analyzed for concentrations of calcium and inorganic phosphorus. Chronograms indicated that the accumulation of both calcium and inorganic phosphorus into the newly forming/least-dense mineral moieties of bone (1.3-1.7 sp grav) showed a single peak in the biorhythm of the rat. A statistically significant circadian rhythm of mineralization was detected for calcium (P less than 0.001) and inorganic phosphorus (P less than 0.039), with peaks during the environmental dark span. These results suggest that the physiological phasing of bone mineralization in the light-dark synchronized rat, is similar to that previously noted for cartilage mineralization and is antiphasal to the midday peak in bone collagen synthesis.     

Effects of Low Temperature, Light and O2 on Chilling-sensitive and -resistant Strains in Chlorella ellipsoidea

A low-temperature sensitive strain, Chlorella ellipsoidea Gerneck(IAM C-102), lost its chilling sensitivity during preservation.Cells of the original strain (low-temperature sensitive) andthe variant (low-temperature resistant) were both synchronouslygrown under a 14-hr light-10-hr dark regime. In the originalstrain, cells at the D-L stage (transient phase) were most sensitiveto a low temperature, whereas the variant cells were not damagedat any stage. During low-temperature treatment, the viability of D-L cellsin the sensitive strain decreased after a lag period of 1 hr.The O₂-uptake activity (respiration) showed the same behavioras the viability, whereas the O₂-evolution activity (photosynthesis)decreased from the start of chilling. In the resistant strain,only O₂ evolution decreased. The decreased activity was restoredwhen the chilled cells were incubated at 25°C. This restorationwas inhibited by oligomycin. Lowering the light intensity or eliminating O2 diminished thechilling injury of the sensitive strain. The results indicatethat the chilling injury of Chlorella results from the combinedeffects of low temperature, light and O₂. (Received September 26, 1980; Accepted March 23, 1981)     

Reorganization and stabilization of acetylcholine receptor aggregates on rat myotubes

Aggregation of acetylcholine receptors (AChRs) is an important early feature of the postsynaptic development of the vertebrae neuromuscular junction. At later stages of differentiation, aggregates are remodeled and stabilized. Aggregation of AChRs can be induced on rat myotubes in culture within 4 hr by treatment with embryonic pig brain extract (EBX). In this study, further sequential changes in the distribution of AChRs were followed by video-intensified fluorescence microscopy. These studies have revealed that groups of AChR aggregates that have formed after 4 hr in EBX are reorganized during the exposure to EBX for 20 additional hr to form a smaller number of larger, oval-shaped aggregates. We have named these two types of aggregates "4-hr aggregates" and "24-hr aggregates". This reorganization occurs by the expansion and merging of individual aggregates within a group, and by the incorporation of newly inserted AChRs. The 24-hr aggregates are an average of 15 times greater in area than 4-hr aggregates, and contain regions with an apparent AChR site density (fluorescence intensity) that is more than twice that of 4-hr aggregates. Electron microscopy of mapped 24-hr aggregates revealed that folded plasma membrane is associated with these regions, probably accounting for the elevated fluorescence. The 24-hr aggregates are more stable than 4-hr aggregates, as determined by their significantly slower disassembly after removal of EBX, elevation of temperature (38 degrees C), reduction of extracellular calcium levels (0.1 mM), or the addition of sodium azide (7 mM). This was determined by following disassembly both statistically (using fixed cultures) and by direct observations of living myotubes. These findings were confirmed by measuring the sequential changes in relative AChR site density over time in individual living myotubes. Thus, 24-hr aggregates form by the reorganization of 4-hr aggregates; exhibit a more regular, compact shape; and are more stable than 4-hr aggregates. These changes in AChR organization and aggregate stability resemble the changes occurring after the initial formation of junctional AChR aggregates during embryonic development, demonstrating additional similarities between this model system and the developing neuromuscular junction.     

Responses of antioxidant enzymes to cold and high light are not correlated to freezing tolerance in natural accessions of Arabidopsis thaliana

Low temperatures and high light cause imbalances in primary and secondary reactions of photosynthesis, and thus can result in oxidative stress. Plants employ a range of low‐molecular weight antioxidants and antioxidant enzymes to prevent oxidative damage, and antioxidant defence is considered an important component of stress tolerance. To figure out whether oxidative stress and antioxidant defence are key factors defining the different cold acclimation capacities of natural accessions of the model plant Arabidopsis thaliana, we investigated hydrogen peroxide (H₂O₂) production, antioxidant enzyme activity and lipid peroxidation during a time course of cold treatment and exposure to high light in four differentially cold‐tolerant natural accessions of Arabidopsis (C24, Nd, Rsch, Te) that span the European distribution range of the species. All accessions except Rsch (from Russia) had elevated H₂O₂ in the cold, indicating that production of reactive oxygen species is part of the cold response in Arabidopsis. Glutathione reductase activity increased in all but Rsch, while ascorbate peroxidase and superoxide dismutase were unchanged and catalase decreased in all but Rsch. Under high light, the Scandinavian accession Te had elevated levels of H₂O₂. Te appeared most sensitive to oxidative stress, having higher malondialdehyde (MDA) levels in the cold and under high light, while only high light caused elevated MDA in the other accessions. Although the most freezing‐tolerant, Te had the highest sensitivity to oxidative stress. No correlation was found between freezing tolerance and activity of antioxidant enzymes in the four accessions investigated, arguing against a key role for antioxidant defence in the differential cold acclimation capacities of Arabidopsis accessions.     

Circadian Rhythm of Blood Ethanol Clearance Rates in Rats: Response to Reversal of the L/D Regimen and to Continuous Darkness and Continuous Illumination

Phase relationships of the circadian rhythms of blood ethanol clearance (metabolic) rates and body temperature were studied in rats successively exposed to 4 illumination regimens: LD (light from 0800-2000 hr), DL (light from 2000-0800 hr), constant darkness (DD) and, lastly, constant light (LL). After a 4-wk standardization to each regimen, body temperatures were taken at 9 × 4-hr intervals to establish baseline circadian profiles. One week later, groups (N = 8) received 1.5 g/kg ethanol (i.p.) at 6 equally spaced timepoints during a 24-hr span, when temperatures were again measured. Ethanol clearance rates were estimated from decreasing blood ethanol levels sampled every 20 min from 60-200 min after dosing, and the resultant elimination curves were subjected to cosinor analysis. These studies show for the first time that the high amplitude circadian rhythm in ethanol metabolism persists under constant conditions of illumination (DD and LL), demonstrating that it may well be a truly internal circadian rhythm and not a response to exogenous cues of the light/dark cycle. During both LD and DL, maximal and minimal ethanol clearance rates fell near the end of the dark and light phases, respectively, and followed circadian peak and trough control temperatures by approximately 6 hr. A fixed internal phase relationship between the core body temperature and the circadian rhythm in ethanol metabolism is demonstrated, thus establishing the rhythm in body temperature as a suitable and convenient internal marker rhythm for studies of the metabolism of low-to-moderate ethanol doses. These studies demonstrate that the phase relationships of blood ethanol clearance rate and body temperature can be manipulated by the illumination regimen selected, an observation of both basic and practical importance.     

Effects of continuous illumination on the rhizome of Caulerpa prolifera

Caulerpa plants were grown under a 12-hr light and 12-hr dark(12L-12D) regime for 8 days followed by 8 days of continuouslight (24L-0D). Under both light regimes the elongation of therhizome was by means of "tip growth". However, the rate of rhizomeelongation in 24L-0D regime (10.9 mm/day) was higher than thatof 12L-12D regime (7.9 mm/day). Jn the region of 1.5 mm fromthe tip, the RERE (relative elemental rate of elongation) under24L-0D and 12L-12D regimes were respectively 4% and 3% elongationper hr. Under 12L-12D photoperiod the subapical part of therhizome exhibited distinctive oscillation: bending upward duringthe light period or at the time of rhizoid cluster initiation,and in the dark becoming relatively straight except at the timeof cluster initiation. No such distinctive oscillation was observedin continuous light. ¹ Report of work supported by research grant GB-7885 from theNational Science Foundation, and in part by Research Councilof Rutgers University. (Received June 8, 1971; )     

Homeostatic and circadian control of body temperature in the fat-tailed gerbil

The interplay of homeostasis and circadian rhythmicity in the control of body temperature was studied in the fat-tailed gerbil (Pachyuromys duprasi). In a first study, the body temperature rhythm of 8 gerbils maintained at 24 degrees C under a 14L:10D light-dark cycle was studied by telemetry. Data from 9 other species of small mammals were also obtained for comparison. The gerbils were found to exhibit a robust rhythm of body temperature (the most robust of the 10 species) with a high plateau during the dark phase of the light-dark cycle and a low plateau during the light phase. In a second experiment, 5 gerbils were allowed to select the temperature of their environment by moving along a thermal gradient. The animals consistently selected higher ambient temperatures during the light phase of the light-dark cycle (when their body temperature was at the low plateau). In a third experiment, the metabolic response of 8 gerbils to an acute cold exposure was determined by indirect calorimetry. Greater cold-induced thermogenesis was observed during the light phase. The fact that the animals selected higher ambient temperatures and displayed greater cold-induced thermogenesis when their body temperature was lower contradicts the hypothesis that the body temperature rhythm is caused by a rhythmic oscillation of the thermoregulatory set point.     

Entrainment of A Free-Running Human with Bright Light?

The case of a 40-year-old sighted woman with free-running sleep-wake and melatonin rhythms is presented. The subject was studied for 102 days. During the pre-treatment period, both the sleep-wake and melatonin rhythms had a period of 25.1 hr, similar to the average period of humans living in temporal isolation. Treatment consisted of bright artifical light exposure (2500 lx Vita-Lite) for 2 hr each day upon awakening. Clock time of light exposure was held constant for 6 days and then slowly advanced until the subject was arising at her desired time of day. The subject continued the light treatment at home and was able to live on a 24-hr day for the 30-day follow-up study. While other factors may be operating in this situation, it is possible that the light treatment caused the stabilization of the free-running rhythms, advancement to a normal phase and entrainment to the 24-hr day. We suspect that the tendency to free-run was related to sleep onsets that were abnormally delayed relative to the circadian phase response curve for light. By scheduling a 2-hr pulse of bright light each morning, this tendency to delay would be counteracted by light-induced advances, resulting in normal entrainment.     

Entrainment of daily serum gonadotropin cycles in the goldfish to photoperiod, feeding, and daily thermocycles

Female fish were kept under 16L:8D/20 degrees C in November and April and the onset of light and/or feeding times were shifted by several hours in the experimental groups. Photoperiod and feeding entrained significant fluctuations in serum gonadotropin hormone (GTH) levels when the onset of light and the first daily feeding were 4 hr apart, but not when they were 10 hr apart. Fish were subjected to 16L:8D for 14-16 days in February, and to either a constant warm (20 degrees C) or a diurnal sinusoidal (12-20 degrees C) temperature regime, the warmth being imposed during photophase or scotophase. While relatively high, uniform serum GTH levels were found throughout the 24-hr period in fish subjected to constant warmth, warm temperature during the day promoted fluctuations in serum GTH levels, and warmth during night resulted in relatively low, uniform serum GTH levels.     

Circannual Variation of Cell Proliferation in Lymphoid Organs and Bone Marrow of Dbf1 Male Mice on Three Lighting Regimens

The case of a 40-year-old sighted woman with free-running sleep-wake and melatonin rhythms is presented. The subject was studied for 102 days. During the pre-treatment period, both the sleep-wake and melatonin rhythms had a period of 25.1 hr, similar to the average period of humans living in temporal isolation. Treatment consisted of bright artifical light exposure (2500 lx Vita-Lite) for 2 hr each day upon awakening. Clock time of light exposure was held constant for 6 days and then slowly advanced until the subject was arising at her desired time of day. The subject continued the light treatment at home and was able to live on a 24-hr day for the 30-day follow-up study. While other factors may be operating in this situation, it is possible that the light treatment caused the stabilization of the free-running rhythms, advancement to a normal phase and entrainment to the 24-hr day. We suspect that the tendency to free-run was related to sleep onsets that were abnormally delayed relative to the circadian phase response curve for light. By scheduling a 2-hr pulse of bright light each morning, this tendency to delay would be counteracted by light-induced advances, resulting in normal entrainment.     

LENGTH OF THE LIGHT-DARK CYCLE AND PLANT GROWTH

Tukey , H. B., Jr ., and H. J. Ketellapper . (California Inst. Tech., Pasadena.) Length of the light-dark cycle and plant growth. Amer. Jour. Bot. 50(2): 110–115. Illus. 1963.—It has been shown that the length of the light-dark cycle which causes maximal growth of tomato, pea, peanut, and soybean plants is close to 24 hr for cycles consisting of equal periods of light and darkness. The exact optimum for tomato plants was determined by temperature; the optimal cycle length was 20 hr at 30 C and 27–30 hr at 14 C. Such an interaction between temperature and cycle length was not found in pea plants, because peas were less sensitive to cycle length than peanuts, tomatoes, and soybeans and did not respond to changes in cycle length of 2–3 hr. The response to cycle length was not influenced by the conditions in which the seedlings had been raised prior to the treatment. Seedlings raised in a 16-hr light, 8-hr dark regime responded in the same manner as those raised in continuous light. The response to cycle lengths of 18, 24, 36, and 48 hr was not changed qualitatively by the temperature during the growth determination. Small changes in cycle length had no characteristic effects on the rates of photosynthesis, respiration or stem elongation. Stem elongation showed a rapid and initial increase in rate when the light was turned off. It was concluded that plants possess an endogenous time-measuring device with a period of 24 hr. For maximal growth to occur the external periodicity must be synchronized with the endogenous period of the plant. Efforts to obtain direct evidence for this hypothesis were not successful since no overt rhythms could be found in tomato plants.     

Hypothermia: teratogenic and protective effects on the development of mouse embryos in vitro

Hypothermia often occurs in association with clinical conditions involving severe hypoglycemia, but its effect on embryonic development has not been well evaluated. Thus, the whole embryo culture method was used to expose day 9 (neurulating) and day 10 (early limb bud stage) mouse embryos to physiologic levels of hypothermia (35 degrees C and 32 degrees C) for 4 and 24 hr. Embryos were evaluated after 24 hours for growth and malformations and compared with controls grown at 37 degrees C. Lactate production was measured in embryos cultured for 4 hr at 32 degrees C and compared with those cultured at 37 degrees C. A 4-hr exposure to hypothermia produced little effect morphologically but reduced the rate of lactate production at both embryonic stages. A 24-hr exposure to hypothermia at 35 degrees C or 32 degrees C produced growth retardation and dysmorphogenesis in embryos undergoing neurulation. Early limb bud stage embryos were less sensitive to this treatment, with growth retardation produced only at the lower temperature. Since hypothermia is commonly associated with severe hypoglycemia in cases of diabetic insulin overdose, day 9 (neurulating) mouse embryos were exposed concurrently to short periods of hypothermia and hypoglycemia and compared with embryos cultured in hypoglycemic medium at normal temperature. The results demonstrate that hypothermia partially protects embryos against the dysmorphogenic effects of hypoglycemia. A balance of metabolic rate and available substrate is discussed as a possible mechanism for this protective effect.     

Nyctohemeral rhythm in melatonin response to isoproterenol in vitro: comparison of rats and Syrian hamsters

The in vitro response of melatonin synthesis was assessed by incubation of individual whole pineal glands for 4 hr without or with different concentrations of isoproterenol in the medium. Pineals were taken either at the end of the 14-hr light phase (day), or at 6 1/2 hr into the 10-hr dark phase (night) after a 30-min exposure of the animals to light just before sacrifice at night. The response was greater in pineals taken at night than in those taken at the end of the light phase in rats, but was absent at the end of the light phase in Syrian hamsters. Hamster pineals taken at night responded, though higher isoproterenol concentrations were required than in the rats. Unresponsiveness of hamster pineals during the day may explain previous failure of isoproterenol administration to stimulate pineal MEL content in this species.     

Atypical 24-hour rhythms of serotonin N-acetyltransferase activity in the rat pineal gland

Previous long-term studies have shown that in the pineal gland of rats melatonin synthesis is subject to infradian rhythms with periods between 4 and 7 days. Since in these studies melatonin-related parameters were measured at one timepoint of a 24-hr cycle only, the aim of the present investigation was to extend these experiments by more frequent sampling, to characterize the infradian rhythmicity in more detail. Male Sprague-Dawley rats kept under a light schedule of LD 12:12 (lights on at 0700) were killed at 6-hr intervals on 8 consecutive days. After decapitation the pineal gland was rapidly dissected out, followed by measurements of one of the melatonin-forming enzymes, serotonin N-acetyltransferase (NAT) activity. It was found that pineal NAT activity exhibited the well known day/night rhythm, i.e. low activity during daytime and strikingly enhanced activity at night, during the first 4 days of the experiment. On the fifth night (from Saturday to Sunday) an unusually high NAT peak occurred at 2400 hr, followed by two atypical 24-hr cycles. In the first cycle the midnight and 0600 hr values were equal and in the second cycle the 0600 hr value was significantly higher than the midnight value. To investigate whether the unusually high NAT peak was a single event or not, four additional short-term experiments were carried out at 2400 hr on 4 consecutive weekends, from Friday to Monday. In each of the four 4-day experiments a distinctly higher peak of NAT activity was found on Saturday, but with time the peaks became less prominent.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of constant darkness and constant light on circadian organization and reproductive responses in the ram

The relationship between circadian rhythms in the blood plasma concentrations of melatonin and rhythms in locomotor activity was studied in adult male sheep (Soay rams) exposed to 16-week periods of short days (8 hr of light and 16 hr of darkness; LD 8:16) or long days (LD 16:8) followed by 16-week periods of constant darkness (dim red light; DD) or constant light (LL). Under both LD 8:16 and LD 16:8, there was a clearly defined 24-hr rhythm in plasma concentrations of melatonin, with high levels throughout the dark phase. Periodogram analysis revealed a 24-hr rhythm in locomotor activity under LD 8:16 and LD 16:8. The main bouts of activity occurred during the light phase. A change from LD 8:16 to LD 16:8 resulted in a decrease in the duration of elevated melatonin secretion (melatonin peak) and an increase in the duration of activity corresponding to the changes in the ratio of light to darkness. In all rams, a significant circadian rhythm of activity persisted over the first 2 weeks following transfer from an entraining photoperiod to DD, with a mean period of 23.77 hr. However, the activity rhythms subsequently became disorganized, as did the 24-hr melatonin rhythms. The introduction of a 1-hr light pulse every 24 hr (LD 1:23) for 2 weeks after 8 weeks under DD reinduced a rhythm in both melatonin secretion and activity: the end of the 1-hr light period acted as the dusk signal, producing a normal temporal association of the two rhythms. Under LL, the 24-hr melatonin rhythms were disrupted, though several rams still showed periods of elevated melatonin secretion. Significant activity rhythms were either absent or a weak component occurred with a period of 24 hr. The introduction of a 1-hr dark period every 24 hr for 2 weeks after 8 weeks under LL (LD 23:1) failed to induce or entrain rhythms in either of the parameters. The occurrence of 24-hr activity rhythm in some rams under LL may indicate nonphotoperiodic entrainment signals in our experimental facility. Reproductive responses to the changes in photoperiod were also monitored. After pretreatment with LD 8:16, the rams were sexually active; exposure to LD 16:8, DD, or LL resulted in a decline in all measures of reproductive function. The decline was slower under DD than LD 16:8 or LL.(ABSTRACT TRUNCATED AT 400 WORDS)     

The Effects of Various Lighting Schedules Upon the Orcadian Rhythms of 23 Liver or Brain Enzymes of C57Bl/6J Mice

The activities of 23 brain or liver enzymes were studied in 5–6 week old C57BL/6JNctr male and female mice that had been fed ad libitum and standardized for 2 weeks to either (1) 12 hr of light (0600-1800) alternating with 12 hr of darkness (1800-0600) (LD 12:12), (2) staggered sequences of 12 hr of light and 12 hr of dark (SLD 12:12) or (3) continuous illumination (LL 12:12) for 2 weeks. Mice in the LD 12:12 and LL 12:12 experiments were killed at 4 hr intervals along a 24-hr span in order to sample at six different circadian stages. Lighting schedules for mice in the SLD 12:12 experiment were organized such that six different circadian stages were sampled when all mice were killed at one time of day.

All 23 enzymes demonstrated a prominent circadian rhythm in at least one of the experiments. Moreover, about two-thirds of the enzymes in LD and SLD 12:12 had a statistically significant fit to a 24-hr cosine curve, while only one-third of the enzymes in LL 12:12 had significant fits to cosine curves. Peak activities of enzymes from mice in LD 12:12 were clustered at the time of transition from light to dark. This was also the trend for the activities of enzymes from mice in SLD 12:12, but resynchronization did not appear completed within the 2-week span. This, along with the observation that mesors (mean 24-hr activity) were reduced and amplitudes altered, indicated that the 2-week standardization period was not sufficient for some enzymes. Times of peak activities, mesors and amplitudes were affected for most enzymes from mice in the LL 12:12 environment. This suggests that individual mice became desynchronized from one another with respect to the original light-dark schedule and that rhythms were altered or lost because individual mice were free running with frequencies different from 24 hr.     

The Effects of Various Lighting Schedules Upon the Orcadian Rhythms of 23 Liver or Brain Enzymes of C57Bl/6J Mice

The activities of 23 brain or liver enzymes were studied in 5-6 week old C57BL/6JNctr male and female mice that had been fed ad libitum and standardized for 2 weeks to either (1) 12 hr of light (0600-1800) alternating with 12 hr of darkness (1800-0600) (LD 12:12), (2) staggered sequences of 12 hr of light and 12 hr of dark (SLD 12:12) or (3) continuous illumination (LL 12:12) for 2 weeks. Mice in the LD 12:12 and LL 12:12 experiments were killed at 4 hr intervals along a 24-hr span in order to sample at six different circadian stages. Lighting schedules for mice in the SLD 12:12 experiment were organized such that six different circadian stages were sampled when all mice were killed at one time of day.

All 23 enzymes demonstrated a prominent circadian rhythm in at least one of the experiments. Moreover, about two-thirds of the enzymes in LD and SLD 12:12 had a statistically significant fit to a 24-hr cosine curve, while only one-third of the enzymes in LL 12:12 had significant fits to cosine curves. Peak activities of enzymes from mice in LD 12:12 were clustered at the time of transition from light to dark. This was also the trend for the activities of enzymes from mice in SLD 12:12, but resynchronization did not appear completed within the 2-week span. This, along with the observation that mesors (mean 24-hr activity) were reduced and amplitudes altered, indicated that the 2-week standardization period was not sufficient for some enzymes. Times of peak activities, mesors and amplitudes were affected for most enzymes from mice in the LL 12:12 environment. This suggests that individual mice became desynchronized from one another with respect to the original light-dark schedule and that rhythms were altered or lost because individual mice were free running with frequencies different from 24 hr.