B. Vivien-Roels

Daily variation in the concentration of melatonin and 5-methoxytryptophol in the human pineal gland: effect of age and Alzheimer's disease

Melatonin and 5-methoxytryptophol (ML) were measured in human pineals (38 controls, 16 subjects with Alzheimers disease). Time of death had a major influence on the indole concentrations with significantly higher melatonin levels occurring at night (22.00-10.00 h) and significantly higher ML levels occurring during the day (10.00-22.00 h). This daily variation disappeared in both the older subjects (55-92 years) and in the Alzheimer patients (55-89 years).

Immunohistochemical evidence for the presence of melatonin in the pineal gland, the retina and the Harderian gland.

SummaryThe presence of melatonin is demonstrated in the pineal gland, the retina and the Harderian gland in some mammalian and non-mammalian vertebrates, using a specific fluorescence labelled antibody technique. Four different potent antibodies against melatonin have been used and compared. In the pineal gland of hamsters, mice, rats and snakes, specific fluorescence, mostly restricted to the cytoplasm of the cells, is detected in pinealocytes. Fluorescence is also detected in the pineal organ of fishes, tortoises and lizards, but it has not been possible, from cryostat sections of fresh tissue, to assert which kind of cell is reacting (photoreceptor cells or interstitial ependymal cells). In the retina, fluorescence is almost exclusively restricted to the outer nuclear layer. In the Harderian gland of mammals and reptiles, fluorescence is localized in the secretory cells of the alveoli and mostly restricted to the cytoplasm surrounding the nucleus. These results are discussed in relation to the concept of melatonin synthesis at extrapineal sites independent of pineal production.

Daily Rhythms of Melatonin Binding Sites in the Rat Pars tuberalis and Suprachiasmatic Nuclei; Evidence for a Regulation of Melatonin Receptors by Melatonin Itself

Using quantitative autoradiography, the density of melatonin binding sites has been measured in the rat pars tuberalis (PT) and suprachiasmatic nuclei (SCN) every 4 h throughout a 24-hour period in animals kept in a light regime of 12L/12D (with lights on at 07.00 h). Slices of PT and SCN were incubated in the presence of 180 and 172 pM, respectively, of 2-125I-melatonin. In both structures investigated, specific 2-125I-melatonin binding sites showed similar rhythms throughout the 24-hour period with a maximum at 16.00 h (PT: 46.9 +/- 2.8 fmol/mg protein, n = 5 and SCN: 5.12 +/- 0.30 fmol/mg protein, n = 5) and a minimum at 4.00 h (PT: 28.5 +/- 4.5 fmol/mg protein, n = 5 and SCN: 3.07 +/- 0.39 fmol/mg protein, n = 5). Similar experiments performed on PT of animals kept in constant light (LL) for 3 days revealed a lack of variations of melatonin binding site density, all the values being significantly higher than those of the respective 12L/12D group (concentration of 2-125I-melatonin used: 180 pM). All these preliminary results were confirmed by saturation studies performed at 16.00 and 4.00 h using quantitative autoradiography and in 12L/12D animals, using radioreceptor binding assays on isolated PT membranes. In 12L/12D animals, the maximum number of melatonin binding sites (Bmax) of both SCN and PT was significantly higher at 16.00 h than at 4.00 h. In all these cases, however, the dissociation constant (Kd) failed to show any significant daily variation.(ABSTRACT TRUNCATED AT 250 WORDS)

Phase-advanced daily rhythms of melatonin, body temperature, and locomotor activity in food-restricted rats fed during daytime.

This study was performed to investigate possible effects of a timed caloric restriction on the light-dark (LD) synchronization of four biological rhythms pair-studied in the same animals. In Experiment 1, food-restricted rats kept under a photoperiod of 12 h light:12 h dark received 50% of previous ad libitum food 2 h after the onset of light. Their daily rhythm of pineal melatonin and rhythms of plasma melatonin and corticosterone were examined and com pared to those of ad libitum control rats after 1 or 2 months of food restriction. A significant phase advance (about 2 h) was found for the pineal melatonin rhythm and for the daily onset of plasma melatonin. Timing of nocturnal peak of circulating corticosterone was unchanged, and a diurnal peak anticipated food presentation by about 2 h. In Experiment 2, effects of a timed caloric restriction under 12L:12D were studied on the expression of daily rhythms of body tem perature and locomotor activity. To discriminate between the effects of timed meal feeding and those of the added caloric restriction, these rhythms were analyzed in food-restricted rats, as in Experiment 1, and were compared to those in sham-restricted rats, concomitantly fed twice more than food-restricted rats (i.e., a timed meal feeding without caloric restriction). Acrophase of the nocturnal peak of body temperature rhythm reached the greatest phase advance (7 h) in food-restricted rats, in which it was close to LD transition. The nocturnal com ponent of locomotor activity rhythm also was markedly phase advanced (6 h) by caloric restriction, as indicated by wheel-running and general activity occur ring from early afternoon to midnight. A smaller 4-h phase advance of the nocturnal peak of body temperature also was observed in sham-restricted rats, although the onset of locomotor activity rhythm apparently was unaffected by meal feeding and the end of activity rhythm was phase advanced by 2 h. These results indicate that timed caloric restriction is a potent phase-shifting agent that interacts with the LD cycle zeitgeber. This nonphotic stimulus phase advances melatonin, corticosterone, body temperature, and activity rhythms to different extents and thus suggests a change in the internal synchronization of the cir cadian system.

Seasonal variation in vasopressin innervation in the brain of the European hamster (Cricetus cricetus)

Using immunocytochemistry, vasopressin innervation was determined in the brain of the European hamster (Cricetus cricetus) during different seasons. It was found that the spring period coincides with a dense vasopressin innervation in many brain regions in the male hamster, and lower vasopressin fibre density in some brain regions in the female hamster. In autumn just before hibernation an almost complete disapperance of vasopressin innervation is noted in those brain regions that are sexually dimorphically innervated in spring. These results suggest that vasopressin activity in certain areas of the brain might be required for some-seasonal functions to find expression.

Daily Variations in Pineal Melatonin Concentrations in Inbred and Outbred Mice

Melatonin was measured using a specific radioimmunoassay in 1 strain of outbred mice (OF1 Swiss) and 4 strains of inbred mice, 2 of them being known to synthesize melatonin (CBA and C3H) and the 2 others being controversial (BALB/c and C57BL/6). In this study, the 5 mouse strains were able to synthesize melatonin, but the basal levels as well as the diurnal variations were very different from one strain to another. CBA and C3H strains showed a clear-cut day-night rhythm of pineal melatonin concentration, with peak levels of 276 ± 22 pg/pineal in CBA and 135 ± 12 pg/pineal in C3H. In BALB/c, the authors confirmed the presence of a very short melatonin peak (15 min) in the middle of the dark period. In C57BL/6 and OF1 Swiss, a very small but significant peak was observed in the middle of the darkness. In the former, another small peak was also observed at light onset. Whether these very small peaks, which may be related to the deficience of N-acetyl transferase activity reported by others, have a physiological meaning remains to be determined.

Effect of environmental temperature and photoperiod on the melatonin levels in the pineal, lateral eye, and plasma of the frog, Rana perezi: Importance of ocular melatonin

Day-night melatonin concentrations were studied in the pineal body, lateral eye, and plasma of the frog Rana perezi in animals maintained in February and July under long (18L:6D) or short (6L:18D) photoperiod and high (25 +/- 1 degree) or low (6 +/- 1 degree) temperature in order to evaluate the influence of these environmental factors. When frogs were kept under short photoperiod and low temperature in February, no melatonin rhythm was observed in the pineal, ocular tissue, and plasma. High temperature at this period of the year induced a day-night rhythm of melatonin levels in the lateral eye and plasma. In July, under long photoperiod and high temperature, animals showed pronounced rhythms of melatonin in the pineal, eye, and plasma. A decrease of environmental temperature in this season abolished the melatonin rhythm. When animals were maintained in August under high (25 +/- 1 degree) temperature and long (18L:6D) or short (6L:18D) photoperiod, the duration of high night time ocular melatonin levels was correlated to the length of the dark phase. In all experiments the high ocular melatonin concentrations and the close parallelism observed between ocular and circulating melatonin profiles suggest that in this species melatonin could be released from the eyes in the general circulation.

Identification of melatonin in the compound eyes of an insect, the locust (Locusta migratoria), by radioimmunoassay and gas chromatography-mass spectrometry

Melatonin, a well-known pineal substance implicated in conveying photoperiodic information in vertebrates, appears also to be present in the compound eyes of an insect, the locust. The identification of melatonin in the eyes of an invertebrate suggests that it may be an evolutionary conservative molecule, principally involved in the time transduction of photoperiodic information in all living organisms.

Overexpression of neuropeptide Y induced by brain-derived neurotrophic factor in the rat hippocampus is long lasting

Brain‐derived neurotrophic factor (BDNF) plays an important role in hippocampal neuroplasticity. In particular, BDNF upregulation in the hippocampus by epileptic seizures suggests its involvement in the neuronal rearrangements accompanying epileptogenesis. We have shown previously that chronic infusion of BDNF in the hippocampus induces a long‐term delay in hippocampal kindling progression. Although BDNF has been shown to enhance the excitability of this structure upon acute application, long‐term transcriptional regulations leading to increased inhibition within the hippocampus may account for its suppressive effects on epileptogenesis. Therefore, the long‐term consequences of a 7‐day chronic intrahippocampal infusion of BDNF (12 μg/day) were investigated up to 2 weeks after the end of the infusion, on the expression of neurotransmitters contained in inhibitory hippocampal interneurons and which display anti‐epileptic properties. Our results show that BDNF does not modify levels of immunostaining for glutamic acid decarboxylase, the rate‐limiting enzyme for γ‐aminobutyric acid (GABA) synthesis, and somatostatin. Conversely, BDNF induces a long‐lasting increase of neuropeptide Y (NPY) in the hippocampus, measured by immunohistochemistry and radioimmunoassay, outlasting the end of the infusion by at least 7 days. The distribution of BDNF‐induced neuropeptide Y immunoreactivity is similar to the pattern observed in animals submitted to hippocampal kindling, with the exception of mossy fibres which only become immunoreactive following seizure activity. The enduring increase of neuropeptide Y expression induced by BDNF in the hippocampus suggests that this neurotrophin can trigger long‐term genomic effects, which may contribute to the neuroplasticity of this structure, in particular during epileptogenesis.

Pineal and circulating melatonin rhythms in the box turtle, Terrapene carolina triunguis: Effect of photoperiod, light pulse, and environmental temperature

Pineal and circulating melatonin concentrations have been measured throughout the 24-hr cycle in the box turtle, Terrapene carolina triunguis, under different conditions of photoperiod and temperature. An obvious effect of photoperiod on the duration of the night rise of pineal and circulating melatonin is observed; the period of elevated melatonin is 4.30 hr in long photoperiod (18L:6D) and 11.00 hr in short photoperiod (8L:16D). A single pulse of 1 hr illumination beginning 1.30 hr after the onset of darkness, in a 16L:8D cycle, has no effect on pineal or circulating melatonin levels. A clear effect of environmental temperature on the amplitude of the day-night rhythm of melatonin production is observed. A possible role of the pineal of poikilotherms in the transduction of several environmental factors, via the daily pattern of melatonin secretion, is hypothesized.