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YVAN TOUITOU
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MICHÈLE FÈVRE
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MICHEL LAGOGUEY
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ALAIN CARAYON
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ANDRÉ BOGDAN
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ALAIN REINBERG
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HERVÉ BECK
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FRANÇOIS CESSELIN
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CATHERINE TOUITOU
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Circadian changes in plasma levels of melatonin, prolactin, LH and FSH were studied in four groups: seven healthy young men, six elderly men, six elderly women and six elderly demented patients (two men and four women). The daily activities of the subjects were synchronous and blood samples were taken every 4 h.

The 24-h mean concentrations of prolactin in plasma were the same in all groups, whereas those of LH and FSH were twice as high in the elderly as in the young men and eight and 23 times higher respectively in the elderly women. The 24-h mean plasma levels of melatonin in the elderly were half those in the young, but were not influenced by the sex or mental condition of the subjects.

A statistically significant circadian rhythm for melatonin was defined in the four groups, for prolactin in all groups except the elderly men and for LH only in the demented patients and in the young men. No circadian rhythm could be detected for FSH in any of the four groups. The acrophases of melatonin and prolactin ranged between 02.30 and 04.00 h, those of LH (when a rhythm was validated) clustered around 01.00 h.

The circadian rhythms of plasma levels of melatonin, prolactin and LH are not modified in old age nor in dementia. A positive correlation has been demonstrated in young men between melatonin and LH and between melatonin and prolactin, but no such correlation could be found in the elderly.

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Yvan Touitou
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José Sulon
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André Bogdan
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Alain Reinberg
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Jean-Claude Sodoyez
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Emilie Demey-Ponsart
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The circannual rhythms of plasma 18-hydroxy-11-deoxycorticosterone (18-OH-DOC), total and free cortisol have been documented on a circadian basis in January, March, June and October in seven young men (24 years old), six elderly men, six elderly women and six elderly demented subjects, both men and women, in their eighties. Blood samples were drawn every 4 h over a 24-h period at each sampling session and urine samples were collected at 4-h intervals only from the young men. A circadian rhythm of 17-hydroxycorticosteroids (17-OH-CS), 17-ketosteroids (17-KS), urinary free cortisol and 18-OH-DOC was defined for each of the four seasons with stable acrophases throughout the year and the same excretory profiles. A circannual rhythm was validated in young men for 17-OH-CS, urinary free cortisol and 18-OH-DOC but not for 17-KS. A circadian rhythm of plasma free cortisol, the active form of the hormone, plasma total cortisol and plasma 18-OH-DOC was validated in all groups and at all the seasons at which samples were taken. The secretory profiles of 18-OH-DOC, free and total cortisol were very similar, with no differences attributable to age, sex or mental condition except for the levels of plasma free cortisol and 18-OH-DOC which were higher and lower respectively in the elderly subjects. Whereas a circannual rhythm of plasma 18-OH-DOC was validated for all groups, a circannual rhythm of both free and total cortisol in the plasma was validated in young men but not in any group of elderly subjects. This loss of the circannual rhythmicity of cortisol in the elderly may reflect the decrease with age of the capacity to adapt to seasonal external factors.

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E. S. Maywood
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M. H. Hastings
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M. Max
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E. Ampleford
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M. Menaker
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A. S. I. Loudon
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ABSTRACT

The aim of this study was to develop a radioimmunoassay for the measurement of endogenous circulating melatonin concentrations in the Syrian hamster, and then to determine the effect of various photic manipulations upon this endocrine signal. In experiment 1, pineal-intact or pinealectomized adult male Syrian hamsters, housed under a long photoperiod (LD; 16 h light:8 h darkness) for 2 weeks, were then either maintained on LD or transferred to a short photoperiod (SD; 8 h light:16 h darkness) for a further 8 weeks. The profile of serum melatonin concentrations was determined from blood samples taken by cardiac puncture at intervals over a 24-h period. Radioimmunoassay revealed that daytime concentrations were at or below the limit of sensitivity of the assay (≥ 50 pmol/l). Under both photoperiods, the concentration of melatonin in the serum of pineal-intact animals rose 4–5 h after the onset of darkness, and the peak amplitude of the melatonin rhythm was not significantly different between the SD- or LD-housed animals (200–250 pmol/l). Premature exposure of animals to light during the dark phase of LD caused a precipitous decline in circulating concentrations to daytime values within 15 min and they remained there for several hours. In animals which experienced an uninterrupted night on either LD or SD, the most striking difference in the rhythm of endogenous melatonin secretion was the duration. Animals housed under LD had high levels until the start of the light period, a peak duration of 3·7 h. In contrast, animals housed under SD exhibited a peak duration of 10 h, levels falling 1·5 h before the start of the light period. The nocturnal increase in serum melatonin concentration was abolished in pinealectomized animals. Serum levels in these animals were not significantly different from those observed in pineal-intact animals during the light phase, being at, or very close to, the limit of sensitivity of the assay.

In experiment 2, animals were housed under LD prior to transfer to continuous darkness for 10 days, during which time their locomotor activity rhythms were recorded. Animals were then chronically cannulated and serial blood samples were removed from the jugular vein at hourly intervals, starting 48 h after surgery. A pronounced circadian rhythm in plasma melatonin concentrations was observed, with levels rising significantly 3 h after the onset of activity and falling 10 h later. During subjective day, levels were at or below the limit of sensitivity of the assay. At the end of the study, pineal melatonin was also measured at intervals across the circadian cycle. This revealed a very close correlation between the plasma and pineal concentrations of melatonin, the duration and phase of both nocturnal peaks being equivalent. These results confirm that the duration of the nocturnal secretion of melatonin varies in proportion to the length of the dark phase, that this rhythm is endogenously driven, and that peak physiological levels of melatonin in this species are of the order of 200 pmol/l.

Journal of Endocrinology (1993) 136, 65–73

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MASAZUMI KAWAKAMI
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JUN ARITA
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Stimulation of the uterine cervix (CS) induced a nocturnal surge of prolactin at 04.00 h and a diurnal surge at 17.00 h in normal ovariectomized rats. However, the CS-induced prolactin surges did not occur in ovariectomized rats which had been treated with 250 μg testosterone propionate during the neonatal period. Chronic bilateral lesions of the suprachiasmatic nucleus (SCN) completely abolished the CS-induced nocturnal and diurnal surges of prolactin release which were observed in sham-lesioned, ovariectomized rats. Furthermore, bilateral lesions of the medial basal part of the suprachiasmatic area (MBSC), lying rostral to the SCN, were also effective in blocking the CS-induced nocturnal and diurnal surges. Lesions which destroyed mainly the optic chiasma and extended partially into the MBSC and SCN did not block the CS-induced prolactin surges.

These results suggest that one reason for the failure of ovary-grafted male rats and neonatally androgenized female rats to maintain pseudopregnancy is the extinction of the circadian rhythm of the two daily prolactin surges, and that the MBSC, in addition to the SCN which is known to be a generator of other circadian rhythms, is involved in generation of the rhythm of prolactin surges.

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Ingram JR
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JN Crockford
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LR Matthews
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Seasonal changes in the activity and responsiveness of the adrenal gland in red deer (Cervus elaphus) stags were quantified by measuring 24 h endogenous cortisol secretory profiles and plasma cortisol responses to either administration of exogenous ACTH or a standardised stressor during November (period of velvet growth), February (pre-rut), April (mid-rut) and July (post-rut) (southern hemisphere) using a remote blood sampling device (DracPac). Ultradian rhythms in the concentration of plasma cortisol were observed resulting from the episodic secretion of cortisol from the adrenal cortex at a mean rate of 0.8 pulses/h. Circadian rhythms in plasma cortisol concentrations were also found in 11 out of the 20 complete 24 h profiles (mean amplitude, 3.8+/-1.4 ng/ml). Seasonal rhythms in mean 24 h plasma cortisol concentrations and cortisol pulse parameters were also observed. Mean 24 h plasma cortisol concentrations were higher in November (12.5+/-1.0 ng/ml) than in February (6.3+/-1.0 ng/ml), April (4.0+/-1.0 ng/ml) or July (4.2+/-1. 0 ng/ml). Cortisol pulse height, nadir and amplitude were all significantly higher in November than at other times of the year (P<0.01). Peak cortisol concentrations following infusion of ACTH(1-24) (0.04 IU kg(-1)) were higher (P<0.05) in November (55.8+/-2.7 ng/ml) and lower (P<0.001) in April (33.7+/-1.8 ng/ml) than those in February and July (48.7+/-2.0 ng/ml and 45.4+/-2.0 ng/ml respectively). The area under the cortisol response curve was significantly smaller (P<0.05) in April (266.6+/-15.3 ng/ml/190 min) than at other times of the year (February, 366.1+/-15.3 ng/ml/190 min; July, 340.7+/-15.3 ng/ml/190 min and November, 387.8+/-21.2 ng/ml/190 min). These data demonstrate that the adrenal gland of the red deer stag exhibits ultradian, circadian and seasonal rhythms in activity, and that its responsiveness to ACTH varies with season. November, a period of reproductive quiescence in the southern hemisphere, with new antler growth and rapid weight gain, is associated with higher mean plasma cortisol concentrations and a greater responsiveness to exogenous ACTH. In contrast, the breeding season is associated with lower adrenal activity and responsiveness.

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A. SZAFARCZYK
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G. IXART
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F. MALAVAL
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J. NOUGUIER-SOULÉ
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I. ASSENMACHER
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SUMMARY

Adrenocorticotrophin (ACTH) and corticosterone in the plasma of adult female rats were measured sequentially at 4 h intervals for 24 h before and after lesions of the suprachiasmatic nuclei or treatment with p-chlorophenylalanine (to inhibit serotonin synthesis). After lesions or p-chlorophenylalanine treatment, the concentrations of ACTH were diminished relative to those in control animals and rhythmic changes could not be detected. However, injection of animals, pretreated with p-chlorophenylalanine, with 5-hydroxytryptophan (60 mg/kg) 8 h before the time when plasma ACTH is maximal in intact animals, stimulated ACTH secretion up to control values. Mean corticosterone concentrations in plasma remained unchanged (after lesions) or increased (after p-chlorophenylalanine). This increase was associated with an increased minimal concentration of corticosterone. After both treatments there was evidence of continued circadian or ultradian rhythms of corticosterone concentration.

Locomotor activity of female rats given identical treatment, but without blood sampling, indicated that nocturnal activity was diminished after lesions whereas diurnal activity was enhanced after p-chlorophenylalanine treatment. Periodicity analysis detected the persistence of free-running circadian, and sometimes ultradian activity, rhythms. Adrenalectomy did not alter further the activity pattern observed in rats with lesions.

These results therefore support the proposition that both the suprachiasmatic nuclei and the serotoninergic system play an irreplaceable role in the mechanism of ACTH secretory rhythms. The suprachiasmatic nuclei are also important for synchronization of locomotor activity and corticosterone rhythms, which may both persist after the suppression of ACTH rhythms.

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Cátia F Gonçalves Wellcome Centre for Cell Matrix Research, Division of Cell Matrix Biology and Regenerative Medicine, School of Biological Sciences, Faculty of Biology, Medicine & Health, Manchester Academic Health Science Centre, University of Manchester, Manchester, UK

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Qing-Jun Meng Wellcome Centre for Cell Matrix Research, Division of Cell Matrix Biology and Regenerative Medicine, School of Biological Sciences, Faculty of Biology, Medicine & Health, Manchester Academic Health Science Centre, University of Manchester, Manchester, UK

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Introduction Circadian (from the Latin circa diem , meaning ‘about a day’) rhythms in behaviour and physiology are a hallmark of life on earth. The 24-h environmental cycles generated by the planet’s rotation around its axis have been wired

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S. Lausson
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N. Segond
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G. Milhaud
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J. F. Staub
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ABSTRACT

The daily changes in rat thyroid calcitonin and its specific mRNA concentrations, and the relationship between their dynamics and the plasma levels of calcitonin, calcium and phosphate over a 24-h period were investigated. The circulating calcitonin concentration rose during the daily dark period when plasma calcium and phosphate levels were minimal, indicating that plasma calcitonin rhythm cannot be generated directly by a linear effect of calcium on hormone secretion. Moreover, we established that the expression of the calcitonin gene also exhibited periodic dynamics observable at the pretranslational level: the gland content of hybridizable specific calcitonin RNA displayed daily rhythms; specific RNA levels peaked during the light period and were minimal during the first part of the dark period. Significant changes in thyroid calcitonin concentrations also occurred over a 24-h period. Statistical analyses which distinguished between variations over the 24-h period and residual variations were performed to test the relationships between the various parameters. The daily rhythms of hybridizable RNA, thyroid calcitonin and plasma minerals appeared to be in phase, while the plasma calcitonin concentration displayed variations out of phase with these rhythms. The implication of the correlations observed on the residual variations is discussed in comparison with the temporal relationship between the daily variations. The results fit the hypothesis that hormone production and secretion are self-oscillating processes. Plasma concentrations of calcium and phosphate might play a role in the synchronization of the calcitonin metabolism periodicity.

Journal of Endocrinology (1989) 122, 527–534

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J. C. BUTTE
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RYOKO KAKIHANA
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E. P. NOBLE
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SUMMARY

In the present study the circadian changes which occur in the levels of corticosterone in the brain and plasma in Sprague–Dawley rats are reported. The levels of corticosterone in the brain were found to have a daily trough and crest with timing similar to that observed for the plasma steroid. In addition, the effect of histamine stress on the corticosterone content of the particulate and the soluble fractions at the trough and crest was examined. The levels of both brain fractions were significantly higher 20 min after histamine injection. The time of day at which the stress was applied was not a significant factor in the magnitude of the stress response.

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Anthony H Tsang Medical Department I, University of Lübeck, Lübeck, Germany

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Mariana Astiz Medical Department I, University of Lübeck, Lübeck, Germany

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Maureen Friedrichs Medical Department I, University of Lübeck, Lübeck, Germany

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Henrik Oster Medical Department I, University of Lübeck, Lübeck, Germany

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physiological target systems according to the time of day. It has long been appreciated that many hormones show circadian rhythms in the circulation ( Pincus et al . 1954 , Moore & Eichler 1972 ). Both central and peripheral tissue clocks impinge on such

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