Takuro Endo

Effects of light and sleep stages on heart rate variability in humans

Effects of light intensity and sleep stages on heart rate variability (HRV) were investigated in young healthy subjects. The low‐frequency (LF)/high‐frequency (HF) ratio was significantly increased by exposing either to bright lights of 10 000 lx or to extreme darkness (< 0.01 lx), while HF and LF components of HRV were not changed, when compared with those under dim light (100 lx). However, LF was significantly increased at REM sleep, when compared with that at the pre‐sleep wake. In contrast, HF was increased at all stages of sleep, and the LF/HF ratio was decreased at slow wave sleep during the baseline night.

Melatonin Receptors in the Spinal Cord

The pineal hormone, melatonin, plays an important role in the regulation of diurnal and seasonal rhythms in animals. In addition to the well established actions on the brain, the possibility of a direct melatonin action on the spinal cord has to be considered. In our laboratory, we have obtained data suggesting that melatonin receptors are present in the spinal cords of birds and mammals. Using radioreceptor binding and quantitative autoradiography assays with 2-[125I]iodomelatonin as the specific melatonin agonist, melatonin binding sites have been demonstrated in the rabbit and chicken spinal cords. These sites are saturable, reversible, specific, guanosine nucleotide-sensitive, of picomolar affinity and femtomolar density. The linearity of Scatchard plots of saturation data and the unity of Hill coefficients indicate that a single class of melatonin binding sites is present in the spinal cord membranes studied. The picomolar affinity of these sites is in line with the circulating levels of melatonin in these animals suggesting that these sites are physiologically relevant. Autoradiography studies in the rabbit spinal cord show that melatonin binding sites are localized in the central gray substance (lamina X). In the chicken spinal cord, these binding sites are localized in dorsal gray horns (laminae I-V) and lamina X. As lamina X and laminae I-II have similar functions, melatonin may have comparable roles in the chicken and rabbit spinal cords. Moreover, in the chicken spinal cord, the density of 2-[125I]iodomelatonin binding in the lumbar segment was significantly higher than those of the cervical and thoracic segments. The densities of these binding sites changed with environmental manipulations. When chickens were adapted to a 12L/12D photoperiod and sacrificed at mid-light and mid-dark, there was a significant diurnal variation in the density (maximum number of binding sites; Bmax) of melatonin binding sites in the spinal cord. After constant light treatment or pinealectomy, the Bmax of melatonin receptors in the chicken spinal cord increased significantly in the subjective mid-dark period. Moreover, there was an age-related decrease in the 2-[125I]iodomelatonin binding to the chicken spinal cord. Our results suggest that melatonin receptors in the chicken spinal cord are regulated by environmental lighting and change with development. These receptors may play an important role in the chronobiology of spinal cord function. The biological responses of melatonin on spinal cords have also been demonstrated in vitro. Melatonin decreased the forskolin-stimulated cAMP production in the chicken spinal cord explant. Preincubation with pertussis toxin blocked the melatonin effect. Our results suggest that melatonin receptors in the chicken spinal cord are linked to the adenylate cyclase via a pertussis toxin-sensitive G protein and that melatonin binding sites in spinal cords are melatonin receptors with biological functions. These receptors may be involved in the regulation of spinal cord functions related to sensory transmission, visceral reflexes and autonomic activities.

Light and Plasma Melatonin Rhythm in Humans

Plasma melatonin rhythm in humans was investigated: its stability, relationship to the sleep-wake rhythm, and response to light. The so-called day-to-day variation of reference phases of plasma melatonin rhythm was within 1.4 h when blood was sampled at 1-hour intervals. Therefore, a change in phase beyond this value is regarded as a phase shift of melatonin rhythm in individuals. Plasma melatonin rhythm was spontaneously desynchronized from the sleep-wake rhythm and probably regulated by the common circadian pacemaker which drives the rhythm in rectal temperature. When a bright-light pulse was applied, the melatonin rhythm produced a phase shift, but the amount of phase shift seems to be different for the ascending and descending phases of nocturnal melatonin rise. Finally, a partial entrainment was observed in a subject who developed a non-24-hour sleep-wake syndrome later, in which the plasma melatonin rhythm was free-running whereas the sleep-wake rhythm was apparently entrained by a 24-hour day-night alternation. It is concluded that the plasma melatonin rhythm is the best marker of the human circadian pacemaker so far available.

Melatonin Excretion Rhythms in the Cultured Pineal Organ of the Lamprey, Lampetra japonica

Pineal organ of the lamprey, Lampetra japonica, is essential to keep the circadian locomotor activity rhythm as previously reported. In this paper, we tried to show that an endogenous oscillator is located and is working in the pineal organ. When the pineal organs were excised and cultured in a plastic tube with M199 medium at 20 degrees C, melatonin secretion rhythms were clearly observed under both light-dark and continuous dark conditions. The circadian secretion of melatonin continued for more than five cycles under the continuous dark condition. This indicates that the pineal organ has an endogenous oscillator and that the melatonin secretion rhythm is controlled by this oscillator. These findings suggest the possibility that the locomotor activity rhythm of the lamprey is under the control of the oscillator in the pineal organ.

Amplitude reduction of plasma melatonin rhythm in association with an internal desynchronization in a subject with non‐24‐hour sleep–wake syndrome

The plasma melatonin rhythm was measured longitudinally in a subject with non‐24‐h sleep–wake syndrome, and the amplitude and area under the curve (AUC) of the melatonin rhythm were investigated in relation to the sleep–wake cycle. When the melatonin rhythm and sleep–wake rhythm were internally desynchronized, the amplitude and the AUC were reduced significantly. These parameters were not influenced by external melatonin administration. These results suggest that a causal relationship between the reduction of circadian oscillation and internal desynchronization exists in this subject.

Influence of κ‐rhythm when assessing drowsiness

Abstract κ‐Rhythm appears at the highest amplitude on a bipolar T3–T4 derivation and its frequency range is 7–10 Hz. When a contra‐lateral earlobe is used as a reference, the κ‐rhythm causes serious problems in assessing drowsiness. Assessment of drowsiness using contra‐ and ipsi‐lateral earlobes as references in 129 subjects who showed κ‐rhythm was compared. Drowsiness could be assessed properly in only 26% of subjects using the contra‐lateral earlobe and in 90% of subjects using the ipsi‐lateral earlobe. These results suggest that the ear on the ipsi‐lateral earlobe should be used as a reference in subjects who show κ‐rhythm.