Duration Of Nocturnal Melatonin Secretion Research Articles

Npas4Is Activated by Melatonin, and Drives the Clock GeneCry1in the Ovine Pars Tuberalis

Seasonal mammals integrate changes in the duration of nocturnal melatonin secretion to drive annual physiologic cycles. Melatonin receptors within the proximal pituitary region, the pars tuberalis (PT), are essential in regulating seasonal neuroendocrine responses. In the ovine PT, melatonin is known to influence acute changes in transcriptional dynamics coupled to the onset (dusk) and offset (dawn) of melatonin secretion, leading to a potential interval-timing mechanism capable of decoding changes in day length (photoperiod). Melatonin offset at dawn is linked to cAMP accumulation, which directly induces transcription of the clock gene Per1. The rise of melatonin at dusk induces a separate and distinct cohort, including the clock-regulated genes Cry1 and Nampt, but little is known of the up-stream mechanisms involved. Here, we used next-generation sequencing of the ovine PT transcriptome at melatonin onset and identified Npas4 as a rapidly induced basic helix-loop-helix Per-Arnt-Sim domain transcription factor. In vivo we show nuclear localization of NPAS4 protein in presumptive melatonin target cells of the PT (α-glycoprotein hormone-expressing cells), whereas in situ hybridization studies identified acute and transient expression in the PT of Npas4 in response to melatonin. In vitro, NPAS4 forms functional dimers with basic helix loop helix-PAS domain cofactors aryl hydrocarbon receptor nuclear translocator (ARNT), ARNT2, and ARNTL, transactivating both Cry1 and Nampt ovine promoter reporters. Using a combination of 5′-deletions and site-directed mutagenesis, we show NPAS4-ARNT transactivation to be codependent upon two conserved central midline elements within the Cry1 promoter. Our data thus reveal NPAS4 as a candidate immediate early-response gene in the ovine PT, driving molecular responses to melatonin.

Pig melatonin receptor 1a (MTNR1A) genotype is associated with seasonal variation of sow litter size

The duration of nocturnal melatonin secretion reaches its minimum in summer, a physiological event that is likely related with the diminished sow fertility and delayed puberty typically observed in this season. Melatonin exerts its function by binding two different receptors named as MTNR1A and MTNR1B. Interestingly, the MTNR1A gene is located on a chromosome SSC17 region where QTL for prolificacy traits have been detected in previous studies. In this work, we have found a synonymous T162C polymorphism at exon 2 of the pig MTNR1A gene. An association analysis between this polymorphism and sow prolificacy in an Iberian × Meishan intercross was performed. The utilization of four statistical models of increasing complexity demonstrated that the MTNR1A gene has both additive and dominant effects on total number of born piglets (TNB) and number of piglets born alive (NBA). Additive effects were significant in summer (TNB, P < 0.01; NBA, P < 0.001), whereas dominant effects reached significance both in fall (TNB, P < 0.01; NBA, P < 0.05) and in winter (TNB, P < 0.001; NBA, P < 0.05). The seasonal variation observed for MTNR1A additive and dominant effects might be produced by the influence of photoperiod on the pattern and duration of melatonin secretion. These results illustrate that the complex interaction between genotype and environment can be an important source of phenotypic variation of reproductive traits.

Thematic review series: Adipocyte Biology. Sympathetic and sensory innervation of white adipose tissue

During our study of the reversal of seasonal obesity in Siberian hamsters, we found an interaction between receptors for the pineal hormone melatonin and the sympathetic nervous system (SNS) outflow from brain to white adipose tissue (WAT). This ultimately led us and others to conclude that the SNS innervation of WAT is the primary initiator of lipid mobilization in these as well as other animals, including humans. There is strong neurochemical (norepinephrine turnover), neuroanatomical (viral tract tracing), and functional (sympathetic denervation-induced blockade of lipolysis) evidence for the role of the SNS in lipid mobilization. Recent findings suggest the presence of WAT sensory innervation based on strong neuroanatomical (viral tract tracing, immunohistochemical markers of sensory nerves) and suggestive functional (capsaicin sensory denervation-induced WAT growth) evidence, the latter implying a role in conveying adiposity information to the brain. By contrast, parasympathetic nervous system innervation of WAT is characterized by largely negative neuroanatomical evidence (viral tract tracing, immunohistochemical and biochemical markers of parasympathetic nerves). Functional evidence (intraneural stimulation and in situ microdialysis) for the role of the SNS innervation in lipid mobilization in human WAT is convincing, with some controversy regarding the level of sympathetic nerve activity in human obesity.

Seasonal Changes in Sleep Duration in African American and African College Students Living In Washington, D.C.

Duration of nocturnal melatonin secretion, a marker of “biological night” that relates to sleep duration, is longer in winter than in summer in patients with seasonal affective disorder (SAD), but not in healthy controls. In this study of African and African American college students, we hypothesized that students who met criteria for winter SAD or subsyndromal SAD (S-SAD) would report sleeping longer in winter than in summer. In addition, based on our previous observation that Africans report more “problems” with change in seasons than African Americans, we expected that the seasonal changes in sleep duration would be greater in African students than in African American students. Based on Seasonal Pattern Assessment Questionnaire (SPAQ) responses, African American and African college students in Washington, D.C. (N = 575) were grouped into a winter SAD/S-SAD group or a no winter diagnosis group, and winter and summer sleep length were determined. We conducted a 2 (season) × 2 (sex) × 2 (ethnicity) × 2 (winter diagnosis group) ANCOVA on reported sleep duration, controlling for age. Contrary to our hypothesis, we found that African and African American students with winter SAD/S-SAD report sleeping longer in the summer than in the winter. No differences in seasonality of sleep were found between African and African American students. Students with winter SAD or S-SAD may need to sacrifice sleep duration in the winter, when their academic functioning/efficiency may be impaired by syndromal or subsyndromal depression, in order to meet seasonally increased academic demands.

Interval Timer Control of Puberty in Photoinhibited Siberian Hamsters

Puberty, which is markedly delayed in male Siberian hamsters (Phodopus sungorus) born into short day lengths, is controlled by an interval timer regulated by the duration of nocturnal melatonin secretion. Properties of the interval timer were assessed by perturbing normal patterns of melatonin secretion in males gestated and maintained thereafter in 1 of 2 short day lengths, 10 h light/day (10 L) or 12L. Melatonin secretion of short-day hamsters was suppressed by constant light treatment or modified by daily injection of propranolol to mimic nocturnal melatonin durations typical of long-day hamsters. Constant light treatment during weeks 3 to 5 induced early incomplete gonadal growth in 12L but not 10 L hamsters but did not affect late onset of gonadal development indicative of puberty in either photoperiod. Propranolol treatment during postnatal weeks 3 to 5 induced transient growth of the testes and ultimately delayed the timing of puberty by 3 weeks. Similar treatments between weeks 5 and 7 or on alternate weeks for 24 weeks did not affect the interval timer. The first 2 weeks after weaning may constitute a critical period during which the interval timer is highly responsive to photoperiod. Alternatively, the hamsters' photoperiodic history rather than age or developmental stage may be the critical variable. The interpolation of long-day melatonin signals at the time of weaning does not appear to reset the interval timer to its zero position but may reduce timer responsiveness to long-day melatonin signals several weeks later.

Decoding the nightly melatonin signal through circadian clockwork

Photoperiod regulates the timing of seasonal cycles in reproduction, energy metabolism, moult and other seasonal characteristics, and the effects are transduced through changes in the duration of nocturnal melatonin secretion from the pineal gland. Short daily melatonin signals (4–8 h/day) activate a summer physiology, while long signals (>10 h/day) produce a winter phenotype. Decoding signal duration occurs in specific target cells in the brain and pituitary gland, each governing a different component of the seasonal adaptation. The pars tuberalis (PT) of the pituitary regulates prolactin release and provides a tractable model system to investigate the molecular decoding mechanism. In the PT, melatonin onset at dusk activates cryptochrone ( Cry1) gene expression and melatonin offset at dawn activates period ( Per1) gene expression, thus the Cry/ Per interval varies directly with nightlength, and inverse to daylength. It is proposed that photoperiod-induced changes in this phase-relationship dictates the level of CRY/PER protein heterodimer formation, and in turn, the level of transcriptional drive to the genes that control PT output – up-regulated under long days stimulating prolactin secretion and a summer physiology, and – down-regulated by short days in winter. The melatonin signal is thus decoded through circadian clock genes.

Independence of circadian entrainment state and responses to melatonin in male Siberian hamsters

BackgroundSeasonal fluctuations in physiology and behavior depend on the duration of nocturnal melatonin secretion programmed by the circadian system. A melatonin signal of a given duration, however, can elicit different responses depending on whether an animal was previously exposed to longer or shorter photoperiod signals (i.e., its photoperiodic history). This report examined in male Siberian hamsters which of two aspects of photoperiod history – prior melatonin exposure or entrainment state of the circadian system – is critical for generating contingent responses to a common photoperiodic signal.ResultsIn Experiment #1, daily melatonin infusions of 5 or 10 h duration stimulated or inhibited gonadal growth, respectively, but had no effect on entrainment of the locomotor activity rhythm to long or short daylengths, thereby demonstrating that melatonin history and entrainment status could be experimentally dissociated. These manipulations were repeated in Experiment #2, and animals were subsequently exposed to a 12 week regimen of naturalistic melatonin signals shown in previous experiments to reveal photoperiodic history effects. Gonadal responses differed as a function of prior melatonin exposure but were unaffected by the circadian entrainment state. Experiment #3 demonstrated that a new photoperiodic history could be imparted during four weeks of exposure to long photoperiods. This effect, moreover, was blocked in animals treated concurrently with constant release melatonin capsules that obscured the endogenous melatonin signal: Following removal of the implants, the gonadal response depended not on the immediately antecedent circadian entrainment state, but on the more remote photoperiodic conditions prior to the melatonin implant.ConclusionsThe interpretation of photoperiodic signals as a function of prior conditions depends specifically on the history of melatonin exposure. The photoperiodic regulation of circadian entrainment state contributes minimally to the interpretation of melatonin signals.

Temperature dependence of gonadal regression in Syrian hamsters exposed to short day lengths.

We sought to determine whether ambient temperature (T(a)) affects gonadal function by altering the rate at which circadian rhythms entrain to short day lengths. Syrian hamsters were housed in cages where they received 14 h of light per day ("long days," 14L) at 22 degrees C. Hamsters were then transferred to cages to receive 10 h of light per day ("short days," 10L) and kept at 5, 22, or 28 degrees C or were maintained in 14L at 22 degrees C. Body mass and estimated testis volume as well as duration of nocturnal locomotor activity (alpha), previously established as a reliable indicator of the duration of nocturnal melatonin secretion, were determined over the course of 24 wk. Testicular regression in short days was accelerated by 4 wk at 5 degrees C and delayed by 3 wk at 28 degrees C relative to 22 degrees C. The interval between alpha-expansion and initiation of testicular regression was markedly affected by T(a) with delays of 0, 3, and 6 wk at 5, 22, and 28 degrees C, respectively. All hamsters held at 5 and 22 degrees C underwent testicular regression, but 25% of those maintained at 28 degrees C failed to do so. We suggest that T(a) modulates testicular regression primarily by affecting responsiveness of neuroendocrine target tissues to long melatonin signals.

A Circadian Signal of Change of Season in Patients With Seasonal Affective Disorder

In animals, the circadian pacemaker regulates seasonal changes in behavior by transmitting a signal of day length to other sites in the organism. The signal is expressed reciprocally in the duration of nocturnal melatonin secretion, which is longer in winter than in summer. We investigated whether such a signal could mediate the effects of change of season on patients with seasonal affective disorder. The duration of melatonin secretion in constant dim light was measured in winter and in summer in 55 patients and 55 matched healthy volunteers. Levels of melatonin were measured in plasma samples that were obtained every 30 minutes for 24 hours in each season. Patients and volunteers responded differently to change of season. In patients, the duration of the nocturnal period of active melatonin secretion was longer in winter than in summer (9.0 +/- 1.3 vs 8.4 +/- 1.3 hours; P=.001) but in healthy volunteers there was no change (9.0 +/- 1.6 vs 8.9 +/- 1.2 hours; P=.5). The results show that patients with seasonal affective disorder generate a biological signal of change of season that is absent in healthy volunteers and that is similar to the signal that mammals use to regulate seasonal changes in their behavior. While not proving causality, this finding is consistent with the hypothesis that neural circuits that mediate the effects of seasonal changes in day length on mammalian behavior mediate effects of season and light treatment on seasonal affective disorder.

Low ambient temperature accelerates short-day responses in Siberian hamsters by altering responsiveness to melatonin.

Exposure to low ambient temperatures (Ta) accelerates appearance of the winter phenotype in Siberian hamsters transferred from long to short day lengths. Because melatonin transduces the effects of day length on the neuroendocrine axis, the authors assessed whether low Ta promotes the transition to winterlike traits by accelerating the onset of increased nocturnal melatonin secretion or by enhancing responsiveness to melatonin in short day lengths. Male hamsters were transferred from 16L (16 h light/day) to 8L (8 h light/day) photoperiods and held at 5 degrees C or 22 degrees C. Locomotor activity was recorded continuously, and body mass, testis size, and pelage color were determined biweekly for 8 weeks. The duration of nocturnal locomotion (alpha), a reliable indicator of the duration of nocturnal melatonin secretion, lengthened significantly earlier in hamsters exposed to a Ta of 5 degrees C than 22 degrees C. Cold exposure increased the proportion of hamsters that were photoresponsive: gonadal regression in short days increased from 44% at 22 degrees C to 81% at 5 degrees C (p < 0.05); low Ta did not, however, accelerate testicular regression in animals that were photoresponsive. Nonphotoresponsive animals at 5 degrees C temporarily had longer alphas during the first 4 weeks in short days and significant decreases in body mass and testicular size that were reversed during the ensuing weeks when alpha decreased. In a 2nd experiment, pinealectomized male hamsters infused for 10 h/day with melatonin for 2 weeks had significantly lower body and testes masses when maintained at 5 degrees C but not 22 degrees C. Low-ambient temperature appears to accelerate the appearance of the winter phenotype primarily by increasing target tissue responsiveness to melatonin and to a lesser extent by augmenting the rate at which the duration of nocturnal melatonin secretion increases in short day lengths.

Establishment and persistence of photoperiodic memory in hamsters.

Long summer days unequivocally stimulate, and short winter days inhibit reproduction in Siberian hamsters. By contrast, intermediate-duration day lengths (12.5-14 h long) either accelerate reproductive development or initiate regression of the reproductive apparatus. Which of these outcomes transpires depends on an animal's photoperiodic history, suggesting that hamsters must encode a representation of prior photoperiods. The duration of nocturnal melatonin secretion is the endocrine representation of day length, but nothing is known about how long it takes to establish photoperiodic histories or how long they endure. Hamsters exposed for 2 or more weeks to long summer day lengths acquired a long-day photoperiodic history that determined subsequent reproductive responses to intermediate-duration day lengths and melatonin signals. The memory for long-day lengths persisted in pinealectomized hamsters for 6.5 weeks, faded significantly after 13 weeks, and was functionally absent after 20 weeks. These findings indicate that hamsters are influenced only by relatively recent day lengths and melatonin signals and ignore earlier ones that might cause them to misinterpret the salience of current day lengths.

Photic entrainment of circannual rhythms in golden-mantled ground squirrels: role of the pineal gland.

Entrainment of circannual rhythms of body mass and reproduction was monitored for 3 years in female golden-mantled ground squirrels maintained in a simulated natural photoperiod. Both pinealectomized and pineal-intact squirrels generated circannual rhythms of body mass and estrus, but only the intact animals entrained these rhythms to a period of 365 days. In the second and third years after treatment, the period of the body mass rhythm was significantly shorter than 365 days for pinealectomized squirrels, and variance in tau among these animals was significantly greater than for intact squirrels. A similar pattern was evident in the rhythm of reproduction, which was phase-disrupted in pinealectomized squirrels but entrained in intacts. Seasonal changes in duration of nocturnal melatonin secretion by the pineal appear to be necessary to produce phase-delays required to entrain the circannual clock to a period of 12 months.

Melatonin and seasonal rhythms.

The pineal hormone melatonin plays a ubiquitous role in biology as a chemical mediator of the effects of season on animal physiology and behavior. Seasonal changes in night length (scotoperiod) induce parallel changes in the duration of melatonin secretion (which occurs exclusively at night), so that it is longer in winter and shorter in summer. These changes in duration of nocturnal melatonin secretion, in turn, trigger seasonal changes in behavior. The retinohypothalamic-pineal (RHP) axis's responses to light are highly conserved in humans. Like other animals, humans secrete melatonin exclusively at night, and they interrupt its secretion when they are exposed to light during the nocturnal period of its secretion. In many individuals, the RHP axis also is capable of detecting changes in the length of the night and making proportional adjustments in the duration of nocturnal melatonin secretion, producing the type of melatonin message that animals use to trigger seasonal changes in their behavior. This has been shown both in naturalistic studies in which melatonin profiles were compared in summer and winter and in experimental studies in which melatonin profiles were compared after chronic exposure to long and short artificial "nights." Individuals who live in modern urban environments differ in the degree to which, or even whether, the intrinsic duration of melatonin secretion (the duration measured in constant dim light) responds to seasonal changes in the length of the solar night. Changes in the intrinsic duration of melatonin secretion that are induced by changes in the scotoperiod are highly correlated with changes in the intrinsic timing of the morning offset of secretion and are only weakly correlated with changes in the intrinsic timing of evening onset of secretion. This finding suggests that differences in the way in which individuals are exposed to, or process, morning light may explain differences in their responsiveness to changes in duration of natural and experimental scotoperiods. Although the human RHP axis clearly is capable of detecting changes in the length of the night and in producing the melatonin message that other animals use to trigger seasonal changes in their behavior, it is not yet known whether or how the human reproductive system or other systems respond to this message.

Natural and melatonin-stimulated changes in the circadian rhythm of prolactin secretion in the ewe during seasonal anestrus.

The relations between the circadian rhythms of melatonin and prolactin, and the effect of melatonin infused into the third ventricle or the mediobasal hypothalamus (MBH) on prolactin secretion and dopamine (DA) release were studied in anestrous ewes under increasing daylength conditions. The decreased amplitude and duration of nocturnal melatonin secretion were accompanied with changes in the daily pattern of prolactin secretion. Marked peaks of prolactin occurred after sunrise and melatonin decreases in February, as well as significant evening or nocturnal peaks under long day conditions between April and July. Melatonin infused into the third ventricle evoked an abrupt increase in the concentration of prolactin after 30 min, and the enhanced prolactin level was significantly higher than during the control infusion (range from 204 +/- 75 to 248 +/- 48 ng/ml vs. 128 +/- 68 to 149 +/- 93 ng/ml, mean +/- SD). The concentrations of DA in MBH perfusates decreased during perfusion of melatonin but to a degree similar to that noted in untreated ewes. These data suggest that short-term infusions of melatonin stimulate the secretion of prolactin in the ewe under increasing daylength conditions, and that this effect is not mediated by changes in DA release.

Bimodal patterns of human melatonin secretion consistent with a two-oscillator model of regulation

In many animals, changes in duration of nocturnal melatonin secretion chemically mediate effects of seasonal changes in nightlength on behavior and physiology. According to one model, the changes in duration of secretion result from adjustments in the timing of two circadian oscillators, one entrained to dusk, controlling onset, and another entrained to dawn, controlling offset. Consistent with this model, in six women, we found separate and reproducible evening and morning peaks of melatonin secretion that might represent the separate expressions of rhythms of two oscillators.

Early-morning administration of short-acting beta blockers for treatment of winter depression

Propranolol, 60 mg or less, was administered daily between 5:30 and 6:00 a.m. to 33 patients with winter depression. After open treatment with a mean dose of 33 mg/day, 24 patients (73%) met the remission criteria; 23 completed double-blind continuation or placebo substitution. Subjects who continued to receive propranolol had a mean increase in Hamilton depression score of 3.5, whereas patients switched to placebo had an increase of 11.2; the difference was statistically significant. These findings are consistent with the hypothesis that duration of nocturnal melatonin secretion is the critical seasonal time cue in humans.

The Durations of Human Melatonin Secretion and Sleep Respond to Changes in Daylength (Photoperiod)

Seasonal changes in daylength (photoperiod) modify the duration of nocturnal melatonin (MT) secretion in many vertebrates. In some cases the changes in MT act as chemical signals that trigger photoperiodic induction of breeding and other seasonal phenomena. It is unclear whether, and to what extent changes in daylength modify the duration of human MT secretion. To address this question, I investigated whether the duration of human MT secretion could be altered by artificial photoperiods. I exposed eight healthy volunteers to a conventional "summer" photoperiod of 16 h light and 8 h darkness for 1 week and to a "winter" photoperiod of 10 h light and 14 h darkness for 4 weeks. As occurs in animals, the duration of nocturnal MT secretion in human beings was longer after exposure to the short photoperiod (12.5 +/- 1.8 vs. 10.3 +/- 0.8 h, t = 3.778, P less than 0.01). The duration of the sleep-phase (recorded by electroencephalogram) was also longer (11.0 +/- 0.8 vs. 7.7 +/- 0.2 h, t = 11.754, P less than 0.001). Whether such changes would lead to significant seasonal changes in human physiology and behavior under natural lighting conditions may be worthy of further investigation.

Photoperiod History, Melatonin, and Reproductive Responses of Male Syrian Hamsters

The action of melatonin (MEL) in mediating photoperiodic history (PPH) effects among male Syrian hamsters was investigated. In Exp. 1, pineal intact males in LD 14:10 received daily injections of MEL (15 micrograms) or ethanol:saline vehicle (SAL) 1 h before lights off for 8 wk to generate two groups experiencing identical photoperiods but distinctly different MEL histories. Following the cessation of injections, males were transferred to either LD 12:12 or LD 8:16 for 8 wk to evaluate whether their reproductive response to the new photoperiod would be more influenced by prior PPH or prior MEL history; MEL history was the significant variable. LD 12:12 caused gradual recrudescence in hamsters that were gonadally regressed following MEL injections. In contrast, LD 12:12 caused gonadal regression in hamsters that had large testes following SAL injections. Exp. 2 evaluated whether PPH influences might be mediated by aftereffects on the period (tau) of the circadian pacemaker regulating many behavioral and physiological rhythms. Pineal intact hamsters were exposed to long or short T cycles consisting of an 8 h photoperiod, repeated every 24.67 h (long T) or 23.33 h (short T) to mimic the aftereffects generated by short or long photoperiods. After 5 wk in these T-cycle conditions, all males were transferred to LD 12:12 for 11 wk. The reproductive response to LD 12:12 was modestly influenced by T-cycle history, even though each T-cycle generated different patterns of entrainment to LD 12:12. These findings support the hypothesis that the response of the reproductive system of male hamsters to an intermediate-duration photoperiod depends upon the duration of nocturnal melatonin secretion associated with hamsters' previous PPH.

Pineal melatonin rhythms and the timing of puberty in mammals.

The direction of change in daylength provides the seasonal time cue for the timing of puberty in many mammalian species. The pattern of melatonin secretion from the pineal gland transduces the environmental light-dark cycle into a signal influencing the neuroendocrine control of sexual maturation. The change in duration of nocturnal melatonin secretion is probably the key feature of the melatonin signal which conveys daylength information. This information may also be used by neuroendocrine axes controlling seasonal changes in pelage colour, growth and metabolism. The mechanism of action of melatonin on neuroendocrine pathways is unknown. Although the ability to synthesize and secrete melatonin in a pattern that reflects the duration of the night may not occur until the postnatal period, the rodent and ovine foetus has the ability to respond in utero to photoperiodic cues to which its mother is exposed in late gestation. Transplacental passage of maternal melatonin is likely to be the mechanism by which photoperiodic cues reach the foetus. Species which do not exhibit seasonal patterns of puberty, such as the human, also secrete melatonin in a pattern which reflects the environmental light-dark cycle, but they do not respond reproductively to the seasonal melatonin information.

Seasonal variation in the daily pattern of plasma melatonin in a wild mammal: the mountain hare (Lepus timidus).

Seasonal differences in daily patterns of plasma melatonin concentration were investigated in both free-living and captive mountain hares, in relation to reproductive activity. There was a marked increase in plasma melatonin concentrations at night at all times of the year. The period of elevation of plasma melatonin above the daytime values was longer in winter than in summer, correlating with the longer duration of darkness. The magnitude of the nighttime rise in melatonin concentrations did not differ significantly between seasons. There was no change in the plasma melatonin profile under similar photoperiods before the summer solstice when hares are sexually active or after the summer solstice when gonadal regression occurs, indicating that melatonin is not directly pro- or antigonadal. These field and laboratory observations support the view that the daily rhythm of melatonin secretion plays a role in the transduction of photoperiodic information in the mountain hare. Furthermore, they favor the hypotheses that the circadian melatonin rhythm transduces photoperiod information by means of the duration of nocturnal melatonin secretion or by the coincidence of elevated melatonin with a particular sensitive period, rather than by the amplitude of the nocturnal rise.