Two-phase response of rat pineal melatonin to lethal whole-body irradiation with gamma rays.

Male Wistar rats adapted to artificial light:dark (LD) regimen 12:12 h were whole-body irradiated with a single dose of 9.6 Gy of gamma rays and sham/irradiated in the night in darkness. The rats were examined 60 min, 1, 3 and 5 days after exposure between 22:00 and 01:30 h in the darkness. The results obtained indicate a two-phase reaction of pineal melatonin after the lethal irradiation of rats: the decline of melatonin concentration early after the exposure (at 60 min) with unchanged serotonin N-acetyltransferase (NAT) activity followed by an increase of melatonin synthesis, accompanied by an increase of pineal and serum melatonin on day 5 after the exposure. NAT activity was increased on day 3 after the exposure. Serum corticosterone concentrations in irradiated rats were increased 60 min and 3 days after exposure. With respect to the antioxidant, immunomodulating and stress-diminishing properties of melatonin, we consider the increase in melatonin synthesis during later periods after irradiation as part of adaptation of the organism to overcome radiation stress.     

Control of hepatic triacylglycerol synthesis. Diurnal variations in hepatic phosphatidate phosphohydrolase activity and in the concentrations of circulating insulin and corticosterone in rats.

Male rats were kept for 14 days with alternating 12h periods of light and darkness. The hepatic activity of soluble phosphatidate phosphohydrolase and the concentration of serum insulin were maximum at about 2h after dark. The peak concentration of serum corticosterone occurred 2h before the dark period. It is proposed that corticosterone is partly responsible for the increased phosphohydrolase activity, and that this enables the liver to increase its capacity to synthesize triacylglycerols during the period of maximum feeding.     

Effect of social isolation on 24-h pattern of stress hormones and leptin in rats

This work analyzes the effect of social isolation of growing male rats on 24-h changes of plasma prolactin, growth hormone, ACTH and leptin, and on plasma and adrenal corticosterone concentrations. At 35 days of life, rats were either individually caged or kept in groups (6-8 animals per cage) under a 12:12 h light/dark schedule (lights on at 08:00 h). A significant arrest of body weight gain regardless of unchanged daily food intake was found in isolated rats after 2 weeks of isolation. On the 4th week, rats were killed at 6 time intervals during a 24-h cycle, beginning at 09:00 h. In isolated rats the 24-h pattern of all parameters tested became distorted, as assessed by Cosinor analysis. When analyzed as a main factor in a factorial analysis of variance, isolation decreased plasma prolactin and growth hormone, increased plasma leptin and corticosterone while decreased adrenal corticosterone. Plasma corticosterone levels correlated significantly with plasma ACTH and with adrenal corticosterone levels in group-caged rats only. These changes can be attributed to an effect of mild stress on the endogenous clock that modulates the circadian hormone release.     

Circadian changes of gluconeogenic enzymes in irradiated rats

The authors studied the effect of whole body irradiation at different times of day on the circadian rhythms of gluconeogenic enzymes. They found that: 1. liver and kidney enzyme activities were highest in the light part of the day and lowest in the middle of the dark part; 2. 12-h circadian rhythm of glucose-6-phosphatase and fructose-1, 6-bisphosphatase activity in the liver and the renal cortex followed a similar course; 3. a lethal whole body dose of 14.4 Gy X-rays did not affect the circadian oscillation curves of the given enzymes, with the exception of fructose-1, 6-bisphosphatase in the liver of irradiated rats, where the rhythm was lost.     

The effect of various photoperiods on daily oscillations of serum corticosterone and insulin in rats.

The effect of various photoperiods on circadian rhythms of chosen parameters was investigated in laboratory rats. SPF male Wistar rats were adapted for six weeks to artificial light-dark cycles (LD 8:16, 12:12, 16:8). The light was switched on at 07.00 h in all regimens. The rats were killed at 3-hour intervals within 24 h, the serum concentration of corticosterone, insulin, glucose, food and water intake was determined. The external and computative acrophases of corticosterone varied in every photoperiod being dependent on the duration of light, the mesor values decreased in LD 16:8 in comparison with other photoperiods. The external acrophase of insulin was located 4 h after light onset in LD 8:16 and 12:12, in LD 16:8 one hour before light onset. The mesor values were approximately equal in all photoperiods. The circadian rhythms of glucose were similar in all regimens. Circadian variation of food and water consumption culminated at the same time in all regimens, the amount of food consumed in light increased with the light duration. Various photoperiods remarkably influenced circadian oscillations of corticosterone and in part food and water intake which could be considered as photoperiodic traits.     

The relation of hepatic in vitro inactivation of corticosteroids to the circadian rhythm of plasma corticosterone.

Plasma corticosterone levels and the in vitro capacity of the liver to reduce the delta(4)3-ketone group of corticosterone were ascertained at 4 hr intervals in male rats maintained on a normal lighting schedule (12L:12D). The rates of delta(4)3-ketone reduction, as well as wet liver weight, were highest during the early portion (08.00 hr) of the light period when liver protein and plasma corticosterone concentrations were low. Shortly (2000 hr) after the beginning of the dark period plasma corticosterone reached peak levels, while hepatic inactivation of corticosterone was markedly depressed. This inverse relationship suggests that the rhythmicity in the capacity of the liver to inactivate corticosterone may contribute to the circadian periodicity of plasma corticosterone.     

Role of photoperiod in reproductive maturation and peripubertal hormone concentrations in male collared lemmings (Dicrostonyx groenlandicus).

Onset of sexual maturation was determined in weanling male collared lemmings exposed to one of three experimental regimens of different photoperiods before and after weaning. Animals gestated in photoperiods of either 16 h light:8 h dark or 8 h light:16 h dark. Those from 16 h light:8 h dark were transferred at 19 days of age to either 20 h light:4 h dark or 8 h light:16 h dark; those gestated under 8 h light: 16 h dark remained in that photoperiod throughout the experiment. After exposure for 15, 20, 25 or 30 days to the postweaning photoperiod, animals were killed and the following parameters assessed: body weight, testes weight, seminal vesicle weight, the presence or absence of epididymal spermatozoa and serum concentrations of prolactin, testosterone and corticosterone. All parameters except serum testosterone were significantly influenced by photoperiod. Animals housed under 8 h light:16 h dark had significantly greater body weights than those housed under 20 h light:4 h dark, a response that differs from that reported for other arvicoline rodents. The group gestated on 16 h light:8 h dark and transferred on day 19 to 8 h light:16 h dark had lower testes and seminal vesicle weights than the other two groups, and mature spermatozoa in the epididymides appeared 5 days later than in the 20 h light:4 h dark group. Serum prolactin was largely undetectable in animals from both 8 h light:16 h dark groups, but all males housed in 20 h light:4 h dark had 2.0-15.0 ng prolactin ml-1. Concentration of serum corticosterone was higher in animals weaned into long photoperiod, and decreased with age. These data indicate that weanling male D. groenlandicus are reproductively photoresponsive, but use a decrease in photoperiod, not static short-photoperiod exposure, to alter the rate of development. Prolactin was largely undetectable in animals exposed to short photoperiod, indicating that high concentrations of this hormone are not important for maturation. Low prolactin concentrations in animals in short photoperiods may mediate the annual moult to white pelage. The short-photoperiod-mediated decrease in corticosterone may play a role in seasonal changes in body weight and composition.     

Circadian rhythm of ketone bodies in the blood of fasting rats

SPF male Wistar rats were kept under standard conditions with a light: dark schedule of 12:12 h. The total ketone body concentration was determined in the blood, and the non-esterified fatty acid level in the serum, of fed rats and of animals which had fasted 24 and 48 h. The amount of ketone bodies in fed rats rose in the second half of the light period and fell with the onset of the dark period. After a 24 h fast, the amount of ketone bodies in the blood rose, but the basic characteristics of the curve and the rhythm remained the same as in fed animals. After a 48 h fast, the mean ketone body concentration was decoupled, a significant phase shift occurred and the rhythm was lost. No relationship between the oscillations of the total ketone body concentration in the blood and the oscillations of the serum non-esterified fatty acid level was found.     

The light cycle as a modulator of aminopyrine demethylation by liver from normal,orchiectomized or adrenalectomized rats

Abstract

As part of studies on the interactive effects of light and endocrine factors on hepatic mixed‐function oxidases, the aminopyrine demethylase activity of liver microsomes and the serum concentrations of corticosterone and testosterone were measured in adult male rats (sham‐operated, castrated or adrenalectomized) exposed for 21 days to one of the following types of environmental lighting: (1) Normal light cyle (L from 09.00 to 21.00 h); (2) Reverse light cycle; (3) Constant light (LL) or (4) Constant darkness (DD). One half of the animals in each of the 12 groups was killed at 06.00 h and the other at 18.00 h on the last day of treatment. A 3‐way analysis of variance, multiple comparisons and correlations allowed the following conclusions: (1) The type of lighting had the most significant effect on demethylase activity, its highest values corresponding to periods of darkness (particularly in the rats exposed to a reverse light cycle) and its lowest to LL. (2) Castrated rats as a group had the lowest demethylase activity. (3) Time of death was a significant factor, particularly for the rats in a reverse light cycle (higher activity at 18.00 than at 06.00 h). (4) There was a significant and positive correlation between aminopyrine demethylase and the rates of 7α‐hydroxylation of 3β‐hydroxy‐5‐androsten‐17‐one previously reported for the same animals. (5) There was no significant correlation between demethylase activity and serum corticosterone or testosterone.     

Circadian oscillations of thyroid hormones and insulin in the serum of fasting rats

Adult male Wistar rats adapted to a 12:12 h light:dark regimen, fed or after a 24- or 48-h fast, were decapitated at 3-h intervals during a single day. They were deprived of food at day-time intervals ensuring that on decapitation they had fasted for the same length of time, i.e. 24 or 48 h. Thyroid hormones, insulin and glucose concentrations were determined in their serum. Fasting did not significantly affect circadian thyroxine, triiodothyronine and reverse triiodothyronine rhythms compared with the findings in fed animals; 24, but not 48 hours' fasting led to a shift in the acrophase of circadian insulin and glucose oscillations compared with fed rats. The maintenance of original circadian thyroid hormones and insulin rhythm in rats which fasted for short lengths of time testifies to a dependence of the stimulus on the time of day.     

Dark-phase light contamination disrupts circadian rhythms in plasma measures of endocrine physiology and metabolism in rats

Dark-phase light contamination can significantly disrupt chronobiologic rhythms, thereby potentially altering the endocrine physiology and metabolism of experimental animals and influencing the outcome of scientific investigations. We sought to determine whether exposure to low-level light contamination during the dark phase influenced the normally entrained circadian rhythms of various substances in plasma. Male Sprague-Dawley rats (n = 6 per group) were housed in photobiologic light-exposure chambers configured to create 1) a 12:12-h light:dark cycle without dark-phase light contamination (control condition; 123 μW/cm(2), lights on at 0600), 2) experimental exposure to a low level of light during the 12-h dark phase (with 0.02, 0.05, 0.06, or 0.08 μW/cm(2) light at night), or 3) constant bright light (123 μW/cm(2)). Dietary and water intakes were recorded daily. After 2 wk, rats underwent 6 low-volume blood draws at 4-h intervals (beginning at 0400) during both the light and dark phases. Circadian rhythms in dietary and water intake and levels of plasma total fatty acids and lipid fractions remained entrained during exposure to either control conditions or low-intensity light during the dark phase. However, these patterns were disrupted in rats exposed to constant bright light. Circadian patterns of plasma melatonin, glucose, lactic acid, and corticosterone were maintained in all rats except those exposed to constant bright light or the highest level of light during the dark phase. Therefore even minimal light contamination during the dark phase can disrupt normal circadian rhythms of endocrine metabolism and physiology and may alter the outcome of scientific investigations.     

High-performance liquid chromatographic separation of corticosteroids in plasma of rats. Its application to the determination of the circadian rhythm of 18-hydroxycorticosterone related to aldosterone and corticosterone

An efficient separation of corticosteroids in plasma of rats was obtained by reversed-phase high-performance liquid chromatography (HPLC). Plasma corticosteroid assays with HPLC separation were used to determine the circadian rhythm of 18-hydroxycorticosterone (18-OHB) and its possible relationship to aldosterone or corticosterone in conscious rats under standard conditions (regular diet; 12-hour light and 12-hour dark cycle). Significant circadian rhythms of plasma corticosterone, 18-OHB and aldosterone were observed with peak values at 20.00 h and nadir values at 08.00 h. The mean ratio of plasma 18-OHB to aldosterone during 24 h was 2.4. The circadian rhythm of 18-OHB was also correlated with that of plasma aldosterone or corticosterone.     

Circadian rhythm and response to an acute stressor of urinary corticosterone, testosterone, and creatinine in adult male mice

In small laboratory species, steroid measures can be obtained more frequently and less invasively from urine than blood. Insofar as urinary levels reflect systemic levels, they could provide advantages particularly for measurement of glucocorticoids, whose blood levels react rapidly to handling and stress. In Experiment 1, urinary samples were collected from male mice every second hour over a 14:10 h light:dark cycle. Samples were analyzed via enzyme immunoassay for corticosterone, testosterone, and creatinine. Corticosterone had peak concentrations 1 h after light offset and a trough 1 h after light onset. Testosterone showed peak concentrations 5-7 h after light onset and lowest concentrations during the dark phase of the cycle. Creatinine showed some variation over the light-dark cycle, but steroid measures showed similar trends with and without adjustment for creatinine. In Experiment 2, mice were stressed via an injection at times close to the determined peak and trough levels of corticosterone. In urinary samples taken 90 min after injection, corticosterone was significantly higher in injected animals at both times relative to levels in control animals, but testosterone was unaffected by injection stress. In Experiment 3, serum and urine samples were collected from mice every sixth hour across the diurnal cycle. Corticosterone peaked in urine and serum immediately after light offset, and urinary measures predicted those in serum. These data indicate that urinary corticosterone reflects systemic levels in mice, document circadian variation in urinary testosterone, and indicate that circadian variation in creatinine is minimal, but potentially relevant in stressed animals.     

The circadian and circa-annual response of rat serum corticosterone to different stressors

The serum corticosterone (B) response to shortlasting immobilization and the administration of ACTH was studied during the year, at the beginning of the light and dark part of the day, in SPF Wistar rats of both sexes kept under standard conditions with a 12:12 h light and dark regimen. Both sexes reacted to the stressors (mainly immobilization) by a marked increase in B at the beginning of the light period. The maximum reaction, to both immobilization and ACTH, was found in males in the spring; females reacted more evenly throughout the year, with a maximum in the winter. The reactivity of both sexes was the lowest in the summer. Circadian and circa-annual differences in the reactivity of the laboratory rat are genetically (evolutionally) fixed and must be taken into account when evaluation the results of experiments.     

Circaseptan rhythm of corticosterone and lipids in the serum of male rats

After adaptation to standard conditions and to a 12:12 h light: dark regimen, groups of young SPF male Wistar rats were killed at 8 h on 17 consecutive days and their serum corticosterone and lipid concentrations were determined. Using a combination of a Fourier harmonic analysis and an analysis of variance, the circaseptan rhythm of corticosterone, total cholesterol and triacylglycerol oscillations was determined; it is unlikely that the nonesterified fatty acid concentration has a 7-day rhythm. The circaseptan variation of metabolic and hormonal indicators is evidently adaptive in character and its existence ought to be taken particularly into account in the evaluation of long-term experiments.     

Acute effects of corticosterone on tissue protein synthesis and insulin-sensitivity in rats in vivo.

The effect of corticosterone treatment on the sensitivity of muscle protein synthesis to insulin infusion was assessed in post-absorptive young rats. To select the optimal time period for corticosterone treatment, protein synthesis was measured by injection of L-[2,6-3H]phenylalanine (1.5 mmol/kg body weight) 1, 4, 12 or 24 h after injection of corticosterone (5 mg/kg body wt.). Muscle protein synthesis was significantly decreased at 4 h and the effect was maximal by 12 h; liver protein synthesis was elevated at 12 h and 24 h. The dose-response of muscle protein synthesis to a 30 min infusion with 0-150 munits of insulin/h was then compared in rats pretreated with corticosterone (10 mg/100 g body wt.) or vehicle alone. When no insulin was infused, corticosterone inhibited protein synthesis in gastrocnemius muscle. High doses of insulin stimulated protein synthesis, but the inhibition by corticosterone was similar to that in the absence of insulin. At intermediate doses of insulin there was an increased requirement for insulin to elicit an equivalent response in muscle protein synthesis. Plantaris muscle responded in a manner similar to that of gastrocnemius, but neither soleus muscle nor liver responded significantly to insulin. These data suggest that corticosterone has two modes of action; one which is independent from and opposite to that of insulin, and a second which causes insulin-resistance through a decrease in sensitivity rather than a change in responsiveness.     

Circadian rhythm of circulating corticosterone and prolactin levels in spontaneously hypertensive rats

Circulating levels of corticosterone and prolactin were measured by radioimmunoassay at hourly intervals during a 24 h period to establish the diurnal rhythm of these hormones in spontaneously hypertensive rats (SHR). Corticosterone levels exhibited a distinct circadian variation with concentrations reaching a zenith at 2000 h and a nadir at 1200 h in male and female SHR. Corticosterone levels in females were consistently greater than males. Circulating prolactin levels were greater during the light h than dark in the female; the opposite occurred in males. Measurement of pituitary prolactin content tended to be low when serum prolactin levels were high and vice versa. The circadian rhythm of circulating corticosterone and prolactin in the hypertensive SHR was found to be similar to the day-night patterns established for normotensive rats. However, these measurements were made under quiescent conditions. It is suggested that because SHR are hyper-responsive to stress and because corticosterone and prolactin have synchronous effects on stress-induced adrenal steroidogenesis, further investigation of prolactin and corticosterone may reveal a participatory role of these hormones in the pathogenesis of the genetically-programmed hypertension of SHR.     

Malondialdehyde content and circadian variations in brain, kidney, liver, and plasma of mice

In aerobic organisms, the use of oxygen (O(2)) to produce energy is associated with the production of Reactive Oxygen Species (ROS), which reacts with biological molecules to produce oxidized metabolites such as malondialdehyde (MDA). This experiment focused on male Swiss mice 12 weeks of age synchronized for 3 weeks by the 12 h light (rest)/12 h dark (activity) span. Different and comparable groups of animals (n=10) were sacrificed at six different circadian stages: 1, 5, 9, 13, 17, and 21 h after light onset (HALO). The 24 h mean MDA level varied among organs of mice in non-stress conditions and was comparable in brain and liver but lower than in kidney. As the MDA 24 h status constitutes only a part of ROS damages in sites differing by their oxygen use, lipid composition, and detoxification capacity, the temporal patterns of their MDA content were comparatively studied in relationship to the animal rest-activity cycle. The results revealed significant circadian rhythms with the peak time located during the rest span (approximately =5 HALO) for both brain and liver, but during the activity span for the kidney ( approximately =21 HALO) and plasma (approximately =13 HALO). This chronobiological study showed that under physiological conditions, lipid peroxidation depends on several factors. The MDA peak/trough might be used as a tool to detect moments of high/low sensitivity of tissues to ROS attack in rodents.     

Circadian characteristics of corticosterone secretion in red-backed voles (Clethrionomys gapperi).

To provide necessary background for study of stress response in red-backed voles (Clethrionomys gapperi), the circadian and ultradian rhythm in corticosterone release was characterized. Animals were maintained under a 16h light, 8h dark cycle. A total of 55 males and 46 females provided 101 independent blood samples over a 6-month span. Samples were obtained at 1h to 2h intervals during the light and at 2h intervals during the dark. Using edited data (5 values beyond the upper 95% limit were removed), a significant time effect was found by analysis of variance (ANOVA) for both sexes at P < .001. The composite single cosine best describing the circadian wave-form for each sex consisted of three components (24h, 12h, and 6h), each significant at P < .05 (overall model P < .001). The 24h mean (mesor) was about 60% higher in females than males (646 ng/mL vs. 412 ng/mL, P = .01), with amplitudes of 429 and 298 ng/mL being proportional (66% vs. 72%) to the respective mesor. The predictable range of change within a 24h span (determined by the double amplitude of a 24h + 12h + 6h cosine model) was large: It was more than 1600 ng/mL for females and more than 900 ng/ mL for males. Highest values were found during the dark phase, with the 24h acrophase located at 2h into the dark span for both sexes. With the caveat of fewer samples obtained during dark than during light, the actual peak values for females occurred at 2h and for males at 6h into the 8h dark span. These results provide baseline information about the circadian time structure for serum corticosterone in red-backed voles under normal light-dark, low-stress conditions.     

Influence of light regimen and the time of year on circadian oscillations of thyroid hormones in rats.

Male SPF rats (Wistar strain) were adapted in the course of the year to natural light (N) and to a 12:12 h (light:dark) artificial light (A) regimen. At approximately the spring and autumn equinox and the summer and winter solstice, rats were killed at 3-h intervals over a 24 h period and their serum thyroxine (T4), triiodothyronine (T3) and reverse T3 levels were determined. The light regimen and time of year significantly influenced the basic characteristics of the oscillations of the hormones. In the N regimen, T4 levels (T3 levels less) culminated in all seasons in correlation to sunrise. In the A regimen they culminated irregularly after daybreak. In animals with the N regimen, the oscillations of the hormones were rhythmic in all seasons, but in the A regimen in only some seasons. In the N regimen, the mean daily T4 concentration value (the mesor) was the highest in the spring and the lowest in the autumn; in the A regimen the mesors were the same, except for a low mesor in the autumn. In both light regimens, the T3 mesors were the highest in the autumn and low in the winter; the rT3 mesors were a mirror image of the T3 mesors. The annual mean of serum T4 concentrations was lower in the N group than in the A group.