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The effect of exogenous melatonin on the adrenocortical axis was studied by determining serum corticosteroid and adrenocorticotrophic hormone (ACTH) levels under basal and stressed conditions in intact, pinealectomized, and adrenalectomized mature male rats. A direct interrelationship between pineal/melatonin and the adrenal cortex, dissociated from the hypothalamic-pituitary axis, was established. Intraperitoneal injection of 100 micrograms melatonin brought about an elevated adrenocortical response recorded in the serum corticosteroid levels 15 hr afterwards. This effect could be demonstrated only at the beginning of the photophase, when physiological levels of corticosteroids and ACTH are at their lowest daily levels. The pattern of this reaction is different from those caused by stressors such as ether or hypoglycemic reagents, and did not occur in pinealectomized animals although the daily rhythm of corticosteroids and ACTH persisted. No effect on ACTH was evident in intact or adrenalectomized rats, where the serum levels remained unchanged. Moreover, the corticotrophin releasing factor (CRF) content of the median eminence was not affected by melatonin.

The distribution of melatonin binding sites in synaptosomal preparations from five brain areas of sham-operated and pinealectomized young male rats (maintained in a 14 h light: 10 h darkness cycle; lights on at 5.00 h) was recorded at 10.00, 18.00 and 24.00 h, 18 days after surgery, using 125I-melatonin as a probe. The densities of 125I-melatonin binding sites in the medulla-pons, hippocampus and hypothalamus of the pinealectomized rats, exhibited clear diurnal variations. However, the densities of binding sites in these brain areas at 18.00 h were lower in the pinealectomized animals than at the other times of day tested, whereas in the sham-operated controls, the binding at 18.00 h was higher than at the other times of day. No diurnal variations were evident in the midbrain and cerebellum of the pinealectomized animals. The apparent affinities of the binding sites toward the ligand in the various brain areas were similar in the pinealectomized and sham-operated animals and did not significantly vary at any of the times recorded. Oral supplementation of melatonin to the rats via drinking water had no effect on the diurnal variations in 125I-melatonin binding in the pinealectomized rat brain. The results indicate that the diurnal variations in 125I-melatonin binding sites in the rat brain are not generated by the pineal but are affected by removal of the gland.

Sham-operated and pinealectomized male rats were maintained at 14 h light:10 h dark cycles (lights-on 5.00 h) and injected daily, for 14 days, with oxazepam or vehicle. 125I-melatonin binding was recorded in synaptosomes prepared at 10.00, 18.00, and 24.00 h from the hypothalamus, hippocampus and medulla-pons of the rats. In the sham-operated, vehicle treated rats, specific 125I-melatonin binding in all brain areas studied was higher at 18.00 h, whereas in the oxazepam-treated animals, binding was higher at 24.00 h than at the other times tested. In the pinealectomized, vehicle-treated rats, the binding recorded at 18.00 h in all three brain areas, was lower than at the other times of day tested. Oxazepam treatment decreased 125I-melatonin binding at 24.00 h in the hippocampus and medulla-pons of the pinealectomized rats and did not significantly affect the binding in the hypothalamus. These results indicate the ability of oxazepam, pinealectomy and their combination, to manipulate the diurnal variations in 125I-melatonin binding sites in the rat brain.

Direct (via the skull) and indirect (via the eyes) light on the rate of adaptation of circadian rhythms of pineal, retina and serum indoleamines and of locomotor activity was examined by observing photoperiod reversal in eye covered and normally sighted chickens. Eye covering did not affect indoleamine levels nor locomotor activity on the regular light:dark (L:D) cycle. However, following photoperiod reversal, the eye covered chickens showed slower rates of adaptation of retinal melatonin and locomotor activity to the new L:D cycle than did the normal sighted chickens. Pineal and serum indole levels were unaffected by eye covering. Our results in birds indicate that 1) light to the eye has a role in governing retinal melatonin and locomotor activity, 2) the pineal is directly photosensitive, and 3) the endogenous rhythm of pineal melatonin may play a role in the entrainment of the locomotor rhythm in the chicken.

Exposure of adult male rats to continuously elevated temperature of 32-34 degrees C caused a significant decrease of HIOMT activity involved in the specific metabolic process of production of melatonin, considered an active pineal hormone. The effect was already evident after 24 h exposure and increased further during the next 48 h. The results obtained substantiate previous data that the pineal gland may be involved in the system regulating adaptation to extreme temperature changes.

Exposure of adult male rats to increased temperature of 33 +/- 1 degrees C for 3 and 10 days brought about decreases in pineal N-acetyltransferase activity. These and previous findings of pineal HIOMT inhibition under similar conditions support the postulation of a possible thermoregulatory role for the pineal gland.

Pinealectomy leads to significantly higher levels of progesterone (on days 19 and 20 of gestation) and oestradiol (on days 21 and 22) in the serum of pregnant rats. These results indicate that during the last phase of pregnancy the pineal gland may be modifying the levels of gonadal hormones, although not affecting the sequence of the physiological events.

After reversing a 12:12-h light-dark regimen of environmental lighting, pineal and retinal melatonin levels of white leghorn chicks recorded at the first mid-darkness were greatly enhanced and reached maximum levels after three more days; those recorded during the light periods indicate gradual decline but were far from the nadir of the original light period, even at the end of the experiment. The first mid-darkness serum melatonin levels recorded after photoperiod reversal were not much different from their original mid-light values. However, on continuing with the reversed regimen, the next mid-darkness levels were sharply increased, and maximum levels were reached after a further 2 days. Under the same experimental conditions light had much more drastic effects, and 6 h on the reversed regimen were sufficient to bring down completely the high value of the original mid-darkness period to the level of the starting mid-light nadir. Retinal N-acetylserotonin (NAS) measured simultaneously had a pattern similar to that of melatonin, but the pineal NAS rhythm did invert completely, albeit gradually. Eye covering did not prevent inversion of pineal and serum melatonin rhythms, which were identical in eye-covered and sighted control chicks on the reversed regimen of light. However, retinal melatonin values of the light periods were significantly less depressed in eye-covered than in sighted control chicks. Moreover, eye covering completely prevented the retinal NAS depression under light but did not affect pineal NAS. During darkness retinal and pineal NAS elevation was sluggish in the eye-covered chicks.

Serum and pituitary LH, prolactin and progesterone changes recorded in parturient rats exposed to continuous darkness throughout pregnancy were not prevented by early ablation of the pineal gland. Similarly, the shorter gestation period observed in rats kept in continuous darkness and the overwhelming tendency of rats in general to deliver during the light period (real or subjective), persisted in the pinealectomized animals. Darkness-induced endocrine changes in rat dams during the perinatal period appear not to be mediated by the pineal gland.

A potentiated production of [14C]melatonin ([14C]Mt) with a simultaneous decline in [14C]serotonin ([14C]5-HT) conversion to its final metabolite [14C]5-hydroxyindole acetic acid ([14C]5-HIAA) was recorded in a rat pineal medium cultured with [14C]tryptophan and either a neuroleptic [haloperidol (HAL) or chloropromazine (CPZ)] or an antidepressant [desipramine (DIP)] drug. A difference between the pharmacological groups of drugs was evident only in [14C]5-HT levels, which were found significantly elevated in the presence of the neuroleptic HAL and reduced in the presence of the tricyclic antidepressant DIP. A parallel trend was observed in norepinephrine (NE)-induced [14C]tryptophan metabolism in the pineal culture medium. All three drugs tested brought about further enhancement of NE-induced synthesis of MT; HAL produced an elevated, but DIP a reduced, level of [14C]5-HT, compared to the NE-induced value.

The effect of TSH, triiodothyronine (T3) and thyroxine (T4) on basic and noradrenaline (NA)-induced indoleamine metabolism was investigated in rat pineal culture based on the observation that, under the experimental system used, melatonin is formed from its amino acid precursor tryptophan. When added to the culture medium, alone or in the presence of NA, TSH in dosages of 5 and 20 mU had no significant effect on pineal tryptophan metabolism.T4 and 2.5 microgram increased the melatonin concentration and, in the presence of NA 10(-4) M, also induced an increase in N-acetylserotonin (NAS). T3, already at a dosage as low as 0.25 microgram enhanced the melatonin concentration, while 2.5 microgram produced significant increases in concentrations of all pineal indoleamines measured. The latter dosage of T3 also enhanced the NA-stimulated NAS. On adding larger quantities of T4 or T3 to the medium none of the changes encountered with the lower dosages could be seen. The results obtained may suggest a direct positive feedback between the thyroid and pineal glands.

The effect of high ambient temperature (34 degrees C) on the function of the female reproductive system, on embryonic development and on outcome of pregnancy, was investigated in heat-exposed sham-operated (Sh) and pinealectomized (Px) golden hamsters maintained under short photoperiod. Plasma prolactin levels were reduced in both heat-exposed groups (ShH and PxH) but pituitary prolactin was increased in the pinealectomized groups irrespective of ambient temperature (21 or 34 degrees C). Pituitary weights and LH contents were not affected in any test group. Heat exposure brought about a reduction in the number of corpora lutea and of pups born, the latter being more drastically reduced in absence of the pineal; the depressant effect of heat on ovarian weight was evident only in the pinealectomized animals. Progesterone levels were not affected in any test group and pregnancy was not prolonged, thus, it would seem that pregnant hamsters adapt themselves well to heat. Moreover, high ambient temperature promoted a rise in pineal. HIOMT activity and boosted cortisol levels in presence of the pineal gland only, which, together with the above findings, shows that the pineal can provide protection for pregnant hamsters against adverse effects of high ambient temperature.

The paper reviews the data accumulated on the interrelationship between the pineal gland and the central nervous system as evidenced by induced neuroanatomical, biochemical and hormonal changes taking place in one system following the activation or inhibition of the other. The possible role of the pineal in modifying behaviour and its involvement in mental impairment is consequently discussed. Further questions dealt with are, whether in addition to the pineal effect via the hypothalamic-pituitary axis on the peripheral non-reproductive glands, thyroid and adrenals, a direct feedback mechanism exists between their hormones and the pineal. The effect of the pineal on the endocrine glands which are known not to be under the direct control of the hypothalamo-pituitary axis, pancreas, thymus and parathyroid, is also discussed.

Exposure of pinealectomized rats to high ambient temperature (35 degrees C; PXH) brought about a diminution in pituitary weight and LH content when compared to their sham-operated peers (35 degrees C) or to pinealectomized controls (22 degrees C). Serum corticosterone level of PXH rats was significantly depressed while heat or pinealectomy alone had no effect. Mean oestrous cycle length was prolonged and blood serum progesterone was increased in the heat-exposed rats. However, the extended oestrous cycles and elevated serum progesterone levels of heat-exposed rats were depressed or abolished by pineal ablation. Thus, the pineal appears to exert a moderating effect on heat-induced endocrine changes in female rats. No changes were noticed in uterine and ovarian weights corrected for body weights either on the day of vaginal opening, at occurrence of the oestrous phase expressed as percentage of total oestrous cycle, or in N-acetyltransferase and hydroxyindole-O-methyltransferase activities.

A thin-layer chromatographic procedure for measuring pineal HIOMT activity is described, based on the methylation of NAS to melatonin. The method enables simple and accurate determination in small aliquots from a single pineal organ homogenate.

Serum LH levels were higher in pinealectomized than in sham-operated control rats during all 4 of the last days of pregnancy studied, although reaching significance only during the final 2 days, 21 and 22. Prolactin (Prl) levels in the serum on the final day of pregnancy, and its contents in the pituitary throughout the entire study period, were significantly lower in pinealectomized rats. Pituitary weights were lower in pinealectomized than in control animals during days 21 and 22 of pregnancy, but no differences were found between the 2 groups in the average number of living foetuses and resorptions. The results presented would seem to indicate that during the last phase of pregnancy the pineal gland plays a role in the modification of gonadotropin synthesis and release. It appears that with LH the release, and with Prl mainly the synthesis, is being affected by the pineal, although the release of Prl may be influenced as well.

Decreased testicular weight with damaged seminiferous tubules were observed in hamsters pinealectomized and exposed continuously to high ambient temperature. In hamsters with intact pineals, exposure to heat resulted in reduced testicular weight without recognizable histological damage. In both groups, serum testosterone levels were elevated, evidently through a compensatory mechanism in the interstitial cells, which did not appear to be damaged. Concomitantly with these findings, reduced serum LH, and pituitary LH and prolactin concentrations were encountered in the heat-exposed pinealectomized hamsters, compared to those of their heat-exposed sham-operated counterparts. These results in hamsters reaffirm a previous postulation of a moderating, protective role for the pineal, against adverse effects of prolonged exposure to heat, in rodents.

The effects of the pineal gland on endocrine function (especially those of the gonads) were investigated in male rats chronically exposed to increased temperature. Weanling male rats were either pinealectomized (Px) or sham-operated (Sh). Following one week of recovery, animals were assigned to either control temperature of 21 +/- 1 degree C (PxC and ShC) or a temperature of 35 +/- 1 degrees C (PxH and ShH). The animals were kept at their respective temperature for at least 30 days. In both groups (PxH and ShH) exposure to increased temperature resulted in a significant reduction in body and hypophysial weights, and in serum LH and testosterone levels as compared with the respective controls (PxC and ShC). Rectal temperature and serum corticosterone were also significantly increased. No changes were found in pineal hydroxyindole-O-methyltransferase activity of sham-operated rats kept in increased temperature (ShH vs. ShC). Pinealectomy alone (PxC vs. ShC) did not alter any of the measured parameters, except for increased pituitary LH content. Increased temperature plus pinealectomy (PxH vs. PxC; and PxH vs. ShH) caused a significant reduction in pituitary LH content and further accentuated (PxH vs. ShH) the diminished serum LH and testosterone levels evoked by exposure to high temperature. The results suggest that in male rats the pineal gland may play a role in moderating the changes in the reproductive processes that are induced by increased temperature.