Swimming depresses nighttime melatonin content without changing N-acetyltransferase activity in the rat pineal gland

Recently, it was shown that a 1.5-ml subcutaneous saline injection depressed N-acetyltransferase (NAT) activity and melatonin content in the rat pineal gland at night. The present studies were undertaken to determine if another perturbation, swimming, could duplicate this response. Rats swam at 23.10 h (lights out at 20.00 h) for 10 min and were killed 15 and 30 min after the unset of swimming. Pineal NAT activity was found to be unaffected while melatonin content was depressed dramatically. Hydroxyindole-O-methyltransferase (HIOMT) activity as well as the content of serotonin (5HT), 5-hydroxytryptophan (5HTP) and 5-hydroxyindoleacetic acid (5HIAA) were not changed by this treatment. In a second study, pineal melatonin again was depressed without a concomitant drop in NAT activity. Mean serum melatonin at 15 min after onset of swimming was increased although the rise was not statistically significant. In the final study, it was found that NAT activity was slightly increased in intact rats and unchanged in adrenalectomized rats at 7 min after swimming onset. At 15 min both intact and adrenalectomized animals had NAT activity values similar to those of controls. Pineal melatonin content in intact and adrenalectomized rats plummeted to 50% of control values at 7 min and fell further to 25% at 15 min. While the rate of melatonin synthesis was not directly measured, lack of change in the activities of the enzymes involved in melatonin synthesis and the contents of two melatonin precursors suggests that swimming depresses pineal melatonin content by enhancing melatonin efflux from the gland.     

Responsiveness of Pineal N-Acetyltransferase and Melatonin in the Cotton Rat Exposed to Either Artificial or Natural Light at Night

In three separate experiments, the effect of acute exposure to either artificial or natural light during darkness of pineal N-acetyltransferase (NAT) activity and melatonin content was studied in the cotton rat (Sigmodon hispidus). The exposure of animals to an artificial-light irradiance of 160,000 microW/cm2 during darkness for either 1 s, 5 s, or 30 min was followed by a precipitous decline in pineal NAT activity and melatonin content when measured at either 15 or 30 min after light onset. When cotton rats were acutely exposed to light at night for 5 s, irradiances of either 3.2, 32, 320, and 3,200 did not suppress either pineal NAT or melatonin 30 min later; however, if the 5-s exposure had an irradiance of either 32,000 or 160,000 microW/cm2, the pineal enzyme activity and indole content were depressed. Moonlight, which had a maximal irradiance of 0.32 microW/cm2, was unable to suppress pineal NAT activity and melatonin content even when the animals were exposed to the moonlight for 30 min. The treatment of cotton rats with either norepinephrine or its agonist, isoproterenol, before their exposure to light at night retarded slightly the suppressive effect of light on the pineal constituents measured. Also, these drug treatments suppressed the pre-exposure levels of both NAT activity and melatonin content in the cotton rat pineal gland.     

Photoreceptor damage and eye pigmentation: influence on the sensitivity of rat pineal N-acetyltransferase activity and melatonin levels to light at night

The threshold of light irradiance capable of inhibiting nighttime pineal serotonin N-acetyltransferase (NAT) activity and melatonin content, and the importance of intact photoreceptors and eye pigmentation on these changes, were investigated in the rat. Groups of intact albino and black-eyed rats and albino animals with light-induced photoreceptor damage were studied in the dark period before, and after 15 and 30 min of exposure to either 0.0005, 0.175 or 3.33 microW/cm2 irradiance of light. In animals with photoreceptor damage, the sensitivity of the pineal gland to light decreased so that only the highest irradiance tested (3.33 microW/cm2) was capable of totally inhibiting pineal NAT activity and melatonin levels. In one study, pineal NAT and melatonin levels in intact albino rats were inhibited by all three irradiances studied. In a second experiment, albino and black-eyed animals behaved identically, only responding with a depression in pineal NAT and melatonin after exposure to light irradiances of either 0.175 or 3.33 microW/cm2. In conclusion, the lowest irradiance of cool white light capable of inhibiting pineal NAT and melatonin in albino rats is around 0.0005 microW/cm2. At the irradiances studied, photoreceptor damage influences the response of pineal NAT and melatonin to acute light exposure at night. On the other hand, eye pigmentation does not seem to have a major effect on the nighttime inhibition of the pineal by light.     

Vasoactive intestinal peptide stimulates N-acetyltransferase and hydroxyindole-O-methyltransferase activities and melatonin production in cultured rat but not in Syrian hamster pineal glands

The purpose of this study was to compare the responses of the Syrian hamster and rat pineal glands in organ culture to vasoactive intestinal peptide (VIP). The endpoints in these studies were the activities of pineal N-acetyltransferase (NAT) and hydroxyindole-O-methyltransferase (HIOMT), as well as pineal and medium melatonin levels. When rat pineal glands were incubated with either VIP (1 microM) or isoproterenol (1 microM), a beta-adrenergic agonist, a significant increase in NAT and HIOMT activities and melatonin levels were observed within 3 hr. Conversely, during the day, VIP (1 microM) was ineffective in stimulating these parameters in hamster pineal gland after incubation times of either 2, 4, 6, or 8 hr. In another experiment, hamster pineal glands were collected from animals killed in the late dark period (after 30 min light exposure). In these glands, isoproterenol promoted NAT activity and melatonin production; however, VIP was ineffective in stimulating either NAT or HIOMT activities; likewise, VIP had no stimulatory effect on pineal melatonin levels at night. Finally, when hamster pineal glands at night were incubated with either 0, 10 nM, 100 nM, 10 microM, or 100 microM VIP, no changes in any parameter of melatonin synthesis were measured. The results indicate that the hamster pineal gland, unlike that of the rat, may not respond to VIP with an increased melatonin production.     

Depression in Rat Pineal N-Acetyltransferase Activity and Melatonin Content Produced by a Hind Leg Saline Injection is Time and Darkness Dependent

It has recently been shown that a 1.5-ml subcutaneous saline injection into the dorsal aspect of the hind limb induces a dramatic and rapid fall in N-acetyltransferase activity and melatonin content of the rat pineal gland at night. Since many studies have shown the opposite response to stress during the day, the first experiment was undertaken to test whether the timing of the saline injection at night influences the response of the pineal gland. In the present studies, rats were kept under light:dark (LD) cycles of 14:10 with lights out daily at 2000 h. Groups of rats were then given a saline injection at one of the following times: 2315, 0015, 0115, 0215, or 0315. Early in the dark phase (2315 and 0015) the saline injection depressed both the N-acetyltransferase (NAT) activity and the melatonin content of the pineal. As the animals were treated later in the dark period, the response became more blunted and, finally, disappeared. In the second experiment, animals that were kept in light during the usual dark period showed no pineal response when subjected to a hind leg injection of saline at either 2315 or 0315. Additionally, no response was seen in the two pineal parameters when rats had darkness onset delayed by 4 h (to 2400) and were then treated with saline at 0410. The results of these studies indicate that the pineal response to saline injection is time dependent. Also, if the nighttime rise in melatonin is suppressed by light exposure, a saline injection has no further inhibitory effect on pineal NAT activity or melatonin levels.     

Stimulatory effect of isoproterenol but not of dibutyryl cyclic AMP on N-acetyltransferase activity and melatonin content of Syrian hamster pineal gland in organ culture

The purpose of this study was to compare the response of Syrian hamster pineal glands in organ culture either to isoproterenol, a beta-adrenergic agonist, or to dibutyryl cyclic AMP. When pineal glands were collected at night, hamsters were exposed to light for 30 min to depress pineal N-acetyltransferase (NAT) activity and melatonin values to low levels. Pineal glands were removed and placed in organ culture containing either isoproterenol or dibutyryl cyclic AMP and subsequent changes in NAT activity and melatonin levels were measured. At night, isoproterenol (10(-7) or 10(-6) M) induced an increase in the NAT activity and melatonin levels in both pineals and culture media. However, dibutyryl cyclic AMP was either ineffective or minimally effective in stimulating these parameters at either different incubation times (2, 4, and 6 h) or drug concentrations (0.1, 0.5, and 1.0 mM). Conversely, when rat pineal glands were incubated with either isoproterenol (10(-7)) or dibutyryl cyclic AMP (0.5 mM) dramatic rises in NAT activity and melatonin levels were observed. In another experiment, hamster pineal glands were collected from animals killed either late in the light period (19.00 h) or in the latter half of the dark period. Isoproterenol promoted NAT activity and melatonin production only in glands collected in the latter half of the dark phase.     

The effects of near-ultraviolet light on serotonin N-acetyltransferase activity in the chick pineal gland

Effects of near-ultraviolet light (UV-A; 325-390 nm, peak at 365 nm) on the activity of the pineal serotonin N-acetyltransferase (NAT; a key regulatory enzyme in melatonin biosynthesis) were examined in chicks. Acute exposure of dark-adapted animals to UV-A radiation produced a marked decline in NAT activity of the pineal gland. The magnitude of this suppression was dependent upon duration of the light pulse and the age of the animals. The decrease in the nighttime NAT activity evoked by a 5 min pulse of UV-A light applied during the fourth hour of the dark phase of the 12 hr light:12 hr dark cycle (LD) gradually deepened during the first 40 min after the return of animals to constant darkness, then the enzyme activity began to rise, reaching control values by 2 hr. Exposure of chicks to a 5 min pulse of UV-A light during the ninth hour of the dark phase produced a marked decline in pineal NAT activity, which was reversible after 15 min of darkness. Pretreatment of animals with an inhibitor of catecholamine synthesis, alpha-methyl-p-tyrosine (300 mg/kg, i.p.), or with a blocker of alpha2-adrenergic receptors, yohimbine (2 mg/kg, i.p.), antagonized the suppressive effect of UV-A light on nighttime NAT activity of the chick pineal gland. It is concluded that UV-A irradiation, similar to visible light, potently suppresses melatonin biosynthesis in the chick pineal gland, with an alpha2-noradrenergic signal playing the role of an intermediate in this action.     

Type II Thyroxine 5''-Deiodinase Activity in the Rat Brown Adipose Tissue, Pineal Gland, Harderian Gland, and Cerebral Cortex: Effect of Acute Cold Exposure and Lack of Relationship to Pineal Melatonin Synthesis

The effect of acute cold exposure for 6 hours on nocturnal type II thyroxine 5'-deiodinase (5'-D) activity was studied in brown adipose tissue (BAT), Harderian gland, cerebral cortex, and pineal gland of the rat. Moreover, the effect of iopanoic acid (IOP), a potent inhibitor of 5'-D activity, on both pineal N-acetyltransferase (NAT) activity and melatonin content in rats maintained in a cold environment was also examined. Results show that acute cold exposure significantly increases 5'-D activity in BAT but not in either the pineal gland, Harderian gland, or cerebral cortex. In all tissues, the injection of IOP reduced dramatically 5'-D activity, while exposure of the animals to light at night reduced 5'-D activity in pineal gland but not in either the Harderian gland or BAT while light exposure at night increased cerebrocortical 5'-D activity. Cold exposure did not change either pineal NAT activity or the melatonin content of the gland. Finally, when pineal 5'-D activity was inhibited by IOP treatment, neither nocturnal pineal NAT activity nor melatonin content was affected.     

N-acetyltransferase is not the rate-limiting enzyme of melatonin synthesis at night

Abstract:  Circadian melatonin production in the pineal gland and retina is under the control of serotonin N-acetyltransferase (NAT) and hydroxyindole-O-methyltransferase. Because NAT activity varies diurnally, it has been considered both the melatonin rhythm-generating enzyme and the rate-limiting enzyme of melatonin synthesis. In rats with dramatically reduced NAT activity due to a H28Y mutation in NAT, melatonin levels remained the same as in wildtype controls, suggesting that NAT does not determine the rate of melatonin production at night. Using a combination of molecular approaches with a sensitive in vivo measurement of pineal diurnal melatonin production, we demonstrate that (i) N-acetylserotonin (NAS), the enzymatic product of NAT, is present in vast excess in the night pineals compared with melatonin; (ii) the continuous increase in NAT protein levels at late night does not produce a proportional increase in melatonin; and (iii) an increase in NAS in the same animal over several circadian cycles do not result in corresponding increase in melatonin output. These results strongly suggest that NAT is not the rate-limiting enzyme of melatonin formation at night.     

Changes in serotonin levels, N-acetyltransferase activity, hydroxyindole-O-methyltransferase activity, and melatonin levels in the pineal gland of the Richardson's ground squirrel in relation to the light-dark cycle

Pineal serotonin and melatonin levels and the activities of hydroxyindole-O-methyltransferase (HIOMT) and N-acetyltransferase (NAT) were studied over a 24-hour period in the pineal gland of the diurnally active Richardson's ground squirrel (Spermophilus richardsonii). Under alternating light-dark conditions (light:dark hours 14:10), pineal serotonin and melatonin levels exhibited a rhythm with high values occurring either during the day (serotonin) or during the night (melatonin). NAT activity was also markedly increased during darkness. HIOMT activity exhibited no 24-hour variation. Exposure of squirrels to constant light for 7 days exaggerated the serotonin rhythm, but obliterated the cycles of NAT and melatonin. Under constant darkness (for 7 days), the rhythms in serotonin, melatonin and NAT persisted, each having a period of about 24 h. In the second study, ground squirrels were exposed to light-dark cycles of either 8:16, 10:14 or 14:10. Under each of these photoperiodic environments, rhythms in pineal NAT and melatonin were apparent. Increasing the daily dark period from 10 to 14 h caused a prolongation of the elevated NAT and melatonin levels. However, a further prolongation of the daily dark period (to 16 h) did not further increase the duration of the rise in NAT and melatonin. The results show that continual light exposure (irradiance of 200 microW/cm2) for 7 days suppresses the pineal rhythms in both NAT activity and melatonin level in the Richardson's ground squirrel. Conversely, light exposure, rather than depressing the serotonin rhythm, actually exaggerates it. Constant darkness for 7 days has little influence on the 24-hour rhythms of either NAT or melatonin.(ABSTRACT TRUNCATED AT 250 WORDS)     

Phototransduction-related circadian changes in indoleamine metabolism in the chick pineal gland in vivo

Abstract: The purpose of this study was to examine the day/night levels of pineal melatonin and its rate limiting enzyme N-acetyltransferase (NAT) in relationship to the ratio of 11-cis-to all-trans-retinal. Three-week-old chicks were placed in 12:12 light: dark (LD 12:12) cycle for one week, pineals were collected during the light phase at 1500 (i.e., after 10 hr light), during the dark phase at 1900 (i.e., 2 hr after dark), at 2100 (i.e., 4 hr after dark), and at 2300 (i.e., 6 hr after dark) and after light extension to 1900. The results show that light-sensitive 11-cis-retinal in the chick pineal has the same diurnal rhythm as NAT and melatonin; all constituents increased within 2 hr of darkness onset (at 1900) and reached their peak after 4 hr of dark. All values were lowest during the light phase at 1500. Low values for 11-cis-retinal, NAT, and melatonin were also seen in the group of chicks which experienced light extension to 1900. The data indicate that in vivo light plays a major role in triggering rhodopsin-bound 11-cis-retinal production within 2–4 hr after darkness onset; this change likely serves as the signal for the subsequent formation of the hormonal product of the pineal gland, melatonin.     

Effect of Magnesium Ions on Rat Pineal N-Acetyltransferase (EC 2. 3 1.5) Activity

N-Acetyltransferase (NAT) activity at night and following isoproterenol treatment in intact animals was higher in rats pretreated with magnesium than in controls. This effect was probably due to enhancement of agonist binding and/or adenylate cyclase activation caused by magnesium ions. A similar effect was observed in vitro, where magnesium increased norepinephrine-stimulated NAT activity in organ-cultured pineal glands, suggesting that magnesium acts primarily on the pineal gland as opposed to some other peripheral or central site. This enhancement of NAT activity by magnesium would presumably lead to increased melatonin production; and as melatonin has been shown to decrease serum magnesium levels, a negative feedback mechanism may exist.     

Non-suppressibility by room light of pineal N-acetyltransferase activity and melatonin levels in two diurnally active rodents, the Mexican ground squirrel (Spermophilus mexicanus) and the eastern chipmunk (Tamias striatus)

The rhythms in pineal N-acetyltransferase (NAT) activity and melatonin levels were studied in the diurnally active Mexican ground squirrel and Eastern chipmunk. In the ground squirrel, both NAT activity and melatonin levels exhibited a marked nocturnal rise; these increases were not prevented by the exposure of these animals to a light irradiance of 200 microW/cm2 throughout the night. In the Eastern chipmunk, darkness at night was also associated with a marked rise in both the activity of the acetylating enzyme as well as the levels of melatonin. Again, these rhythms were not suppressed by the exposure of these animals to a light irradiance of 200 microW/cm2 for either 1 night or for 7 nights; exposure of chipmunks to light for 7 consecutive days did, however, reduce the rise in melatonin normally associated with darkness. The non-suppressibility of pineal NAT and melatonin by a 200 microW/cm2 light irradiance may relate either to the activity pattern of these animals, i.e., diurnal, or to their previous lighting history.     

Estrogen modulates the nocturnal synthesis of melatonin in peripubertal female rats

Abstract: Our objective was to evaluate the effects of estrogen deficit and of estrogen stimulation on the synthesis of pineal melatonin in female rats during the peripubertal period. The levels of melatonin and N-acetylserotonin (NAS) and the activities of N-acetyltransferase (NAT) and hydroxyindole-O-methyltransferase (HIOMT) were determined in homogenates of pineal glands obtained from peripubertal female Sprague- Dawley rats 4 to 12 weeks of age in the mid-dark during the daily light/dark cycle. Animals were ovariectomized at 4 weeks of age; daily administration of estradiol benzoate (E₂B, 1.0 μg/d, s.c.) was initiated at 4 weeks of age. A peak in the pineal levels of melatonin and NAS and in NAT activity was observed in untreated (control) rats with intact ovaries at 6 weeks. HIOMT activity increased from Week 4 to 6 and remained unchanged thereafter. Ovariectomy at Week 4 led to significant increases in the levels of melatonin and of NAS and NAT in activity at Week 8. NAT activity Week 10 resembled that of control animals, but levels of melatonin and NAS were slightly elevated. Ovariectomy did not affect HIOMT activity. Subcutaneous injection of E₂B significantly decreased the levels of melatonin and NAS and of NAT activity at Week 4, as compared with those in control rats. E₂B suppressed the ovariectomy-induced elevation of levels of melatonin and NAS and of NAT activity, similar to the effect in control animals. E₂B did not affect HIOMT activity. Our results suggest that estrogen modulates the nocturnal synthesis of melatonin in the pineal gland in peripubertal female rats. The effects of estrogen on melatonin synthesis appeared to be mediated by the modulation of NAT activity.     

Light-induced changes in pineal gland N-acetyltransferase activity: developmental aspects

In adult rats, light acting via a retino-pineal gland neural pathway influences pineal gland biochemistry in two ways: (1) it entrains endogenous circadian rhythms in melatonin biosynthesis to the environmental photoperiod and (2) exposure to even very brief periods of light during the nighttime rapidly suppresses the high levels of nocturnal melatonin production. The present studies were undertaken to determine precisely when photic stimulation first influences the enzymic activity of N-acetyltransferase (NAT), the pineal gland enzyme which rate-limits the overall biosynthesis of the hormone melatonin, and to examine some of the cellular mechanisms which might mediate light-induced effects in neonatal animals. Rats of different ages were either killed during the light phase or were exposed to darkness or light for 1 min during the dark phase of the lighting cycle, returned to their litters in darkness for 30 min and then killed. Pineal gland NAT activity in animals nocturnally exposed to 1 min of light was suppressed in animals 6 days of age or older. Nocturnal light exposure did not suppress enzyme activity in 3- to 5-day-old rats, even though these animals displayed clear light:dark differences in pineal gland NAT activity. Nocturnal light exposure also did not suppress nighttime levels of NAT activity in 7-day-old animals who had been bilaterally enucleated, suggesting that this effect is retinally mediated. Pretreatment of 7-day-old animals with the beta-noradrenoceptor agonist drug, isoproterenol, prevented the nocturnal light-induced suppression of NAT activity.(ABSTRACT TRUNCATED AT 250 WORDS)     

Day/night serotonin levels in the pineal gland of male BALB/c mice with melatonin deficiency

Studies from another laboratory have shown that several strains of laboratory mice have a genetic defect for melatonin synthesis. In non-deficient species, melatonin synthesis undergoes a typical, beta-adrenergically regulated day/night rhythm with low melatonin levels during daytime and high levels at night, the precursor serotonin showing an inverse behaviour. This study examines whether a day/night rhythm of pineal serotonin levels exists in melatonin-deficient male BALB/c mice. Mice kept under a lighting schedule of 12 h light (lights on at 07.00 h) and 12 h dark were killed at 13.00 and 01.00 h, respectively. Serotonin amounted to 12-15 ng/pineal and did not show regular day/night differences. Administration of the beta-adrenergic agonist, isoproterenol, which is known to affect melatonin synthesis in a number of species, was without effect on pineal serotonin levels. Melatonin and two of the melatonin-forming enzymes, serotonin N-acetyltransferase (NAT) and hydroxyindole-O-methyltransferase (HIOMT) were below the detection limit in the pineal. It is concluded that in melatonin-deficient BALB/c mice, pineal serotonin synthesis is apparently intact. In BALB/c mice, serotonin synthesis and release do not appear to be directly or indirectly regulated by beta-adrenergic mechanisms.     

Effect of Acute Light Exposure Upon Melatonin Content, NAT Activity, and Nuclear Volume in the Gerbil Pineal Complex

This report describes the response of both the superficial and deep components of the gerbil pineal gland to a 30-min pulse of fluorescent light at the time of the nocturnal peak in pineal activity. When gerbils were maintained in a 14 h light:10 h dark photoperiod, mean melatonin concentrations in the superficial pineal gland ranged between 72 and 108 pg/gland during the day and first 4 h of darkness, rose to a peak of 399 pg/gland 7 h after lights out, then returned to basal levels at the expected time of light onset. Pineal melatonin contents were reduced to daytime values within 30 min after exposure of gerbils to fluorescent light at the time of the nocturnal peak. These changes in superficial pineal melatonin content reflected the circadian pattern in serotonin N-acetyltransferase (NAT) activity, as would be expected if NAT controls melatonin synthesis. In addition to the reductions in NAT and melatonin content induced by nocturnal photic stimulation, there was a 30% reduction in mean pinealocyte nuclear volume. A nocturnal elevation in melatonin content but not in NAT activity occurred in the deep pineal gland. Unlike the superficial pineal gland, the deep pineal gland did not demonstrate a response in melatonin content, NAT activity, or pinealocyte nuclear volumes upon exposure of gerbils to light at the time of the nocturnal peak in superficial pineal activity.     

Electrical stimulation of the hypothalamic paraventricular nuclei inhibits pineal melatonin synthesis in male rats

Recent findings have shown that lesions of the hypothalamic paraventricular nuclei (PVN) disrupt the synthesis of melatonin in the rat pineal gland. In order to further clarify the role of the PVN in the control of pineal function, the effects of electrical stimulation of these nuclei were investigated in acutely blinded adult male Sprague-Dawley rats. Following electrical stimulation, pineal serotonin-N-acetyltransferase (NAT) activity and pineal melatonin content were measured by means of radioenzymatic and radioimmunoassay methods, respectively. Stimulation had no significant effect on pineal melatonin synthesis throughout the early part of the dark phase, but caused a significant reduction in NAT activity during the light phase and the latter part of the dark phase. The pineal melatonin content appeared reduced, but due to large individual variations this reduction was not statistically significant. Stimulation duration experiments reveal that reduction of NAT activity is time dependent, with significant inhibition occurring after 30 min of stimulation. These observations further support the involvement of the PVN in the melatonin rhythm generating pathway and suggest that electrical activation of fibers in the PVN is similar to the effects of light on pineal melatonin synthesis.     

Tryptophan administration inhibits nocturnal N-acetyltransferase activity and melatonin content in the rat pineal gland. Evidence that serotonin modulates melatonin production via a receptor-mediated mechanism.

In the rat pineal gland, the activity of serotonin N-acetyltransferase (NAT) and the concentration of melatonin are normally high at night; conversely, the concentration of serotonin (5-HT), the precursor of melatonin, is low. Since tryptophan administration increases the concentration of pineal 5-HT at night, we examined its effect of melatonin production. Nighttime tryptophan loading led to substantial increases in pineal 5-hydroxytryptophan, 5-hydroxyindole acetic acid (5-HIAA), and 5-HT but a highly significant reduction in NAT activity in comparison to saline-injected controls. In contrast to other measured indoles, melatonin levels also were significantly diminished by tryptophan loading. Nocturnally high pineal norepinephrine levels were unaltered by tryptophan administration. The idea that high concentrations of 5-HT could lead to substrate inhibition of NAT activity was not supported by kinetic analysis of control NAT levels versus tryptophan-inhibited NAT activity under varied substrate concentrations. Hypotheses to explain these results include the possibility that tryptophan inhibition of melatonin synthesis is mediated by the release of 5-HT from the pinealocyte and its subsequent autocrine action on melatonin production.     

Changes in nocturnal pineal melatonin synthesis during the perimenopausal period: Relation to estrogen levels in female rats

To evaluate changes in melatonin synthesis during the perimenopausal period in the female rat and to determine the effects of estrogen on melatonin synthesis, pineal levels of tryptophan, melatonin and norepinephrine and activities of N-acetyltransferase (NAT) and hydroxyindole-O-methyltransferase (HIOMT) were determined. Homogenates for assay were prepared from the pineal glands of female virgin Sprague-Dawley rats between 4 and 24 months of age in the middle of the dark period of a daily light/dark cycle. Serum 17 beta-estradiol (E2) concentrations were also determined. Pineal melatonin levels significantly decreased from month 4 12 and significantly increased from month 12 16, decreasing thereafter. Serum E2 concentrations significantly decreased from month 12-16, and remained low thereafter. No significant changes in tryptophan or norepinephrine were seen. NAT activities paralleled the time course of changes in melatonin. HIOMT activities decreased gradually from month 4 24. Subcutaneous implantation of an E2 capsule between months 12 and 16 resulted in significant decreases in levels of melatonin and NAT activity at month 16. Ovariectomy at month 4 or 12 led to significant increases in the levels of melatonin and NAT activity. These findings represent a temporal increase in pineal melatonin synthesis during the perimenopausal period, and suggest that the increase in melatonin synthesis activity at that time might result from decreasing levels of endogenous estrogen. The effect of estrogen on melatonin synthesis appeared to involve modulation of NAT activity.