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A clinical study of the pineal gland activity in oncologic patients.

It has been known for many years that the pineal gland is involved in regulating tumor growth. In order to evaluate the functional activity of the pineal gland in neoplastic diseases, melatonin serum levels and its light/dark rhythm have been determined with the RIA method in patients affected by various forms of tumor. Irrespectively of the type of the tumor and of its localization, existence of two subpopulations has been observed within the oncologic patients, the former with normal levels of melatonin, and the latter with high ones. The light/dark rhythm of melatonin was anomalous in some cases. An evident decrease of serum melatonin values was seen after chemotherapy. It might be interesting to establish whether melatonin levels may conditionate the prognosis of patients with cancer.

Adolescent↗

The effects of constant and diurnal illumination of the pineal gland and the eyes on ocular growth in chicks.

PURPOSE: To investigate the effects of constant or 12-hour cyclic illumination of the pineal gland and the eyes on the growth of the chick eye. METHODS: Chicks (Gallus gallus, Cornell K Strain) were raised either under a 12-hour light-dark cycle of normal light or under constant light, with or without opaque removable hoods that covered the top of the head for 12 hours each day. A second group of chicks was raised under constant light with opaque eye covers that were worn on either both eyes or only the right eye for 12 hours each day. Chicks were placed in the experimental conditions on the third day after hatching and raised for 3 weeks. RESULTS: Pineal gland hoods and eye covers worn 12 hours a day significantly (P < 0.0001) protected the chicks from hyperopia under constant-light conditions. They also reduced the flattening of the cornea caused by constant light. Most striking was the protection afforded the uncovered eye from constant light's effects by the periodic covering of the opposite eye. CONCLUSIONS: A diurnal light-dark rhythm presented to one of three photosensitive organs (the pineal gland and both eyes) can protect the eyes from the effects of constant light. This is most probably due to the maintenance of a melatonin rhythm in the organ receiving the diurnal light rhythm.

Animals↗

Phosphoinositide metabolism of rat pineal gland in a low ionic strength medium.

Pineal glands were incubated in the presence of 32P orthophosphate. When all NaCl in a conventional incubation medium was replaced by isotonic sucrose, i.e. when the ionic strength of the medium was decreased, there was a marked increase in 32P labelling of phosphatidylinositol (PI) and phosphatidic acid (PA). The 32P labelling of phosphatidylcholine (PC) and phosphatidylethanolamine (PE) was not affected. No net synthesis of PI was observed. The increased labelling of PI therefore represents an increase in the turnover of PI. The 32P labelling of PI was observed also in media where NaCl was replaced by fructose or mannitol, but not in media, where NaCl was replaced by choline chloride. The effect depends on the concentration of the HEPES buffer and was not found in the medium with a bicarbonate buffer. 32P labelling of PI was not blocked by alpha 1 adrenergic blockers, phentolamine and prazosin, and did not depend on the presence of Ca2+ in the incubation medium. The effect was blocked by a Ca2+ channel blocker, MnCl2. Only 32P labelling of PI and not that of phosphatidylinositol 4-phosphate (PIP) and phosphatidylinositol 4,5-bisphosphate (PIP2) was increased during prolonged incubation in the sucrose medium. It is suggested that a decrease in the charge distribution across the plasma membrane as a result of the absence of most monovalent cations is responsible for the increased metabolism of phosphatidylinositol.

Animals↗

Expression of class II MHC molecules in the rat pineal gland during development and effects of treatment with carbon tetrachloride.

Cells expressing major histocompatibility complex (MHC) class II (Ia) antigen have been examined during the development of rat pineals and in the pineal gland of adult rats treated with carbon tetrachloride. Cells positive for MHC class II are first detected in the pineal gland of the 7-day-old rat. These positive cells increase in number gradually during development, MHC class II immunoreactivity reaching adult levels at 4 weeks after birth. The MHC class II antigen is intensely labeled on the cell surface, and labeled cells are distributed throughout the organ, several positive cells being gathered into groups. The positive cells are small (7-12 microm in diameter), irregular in shape, and frequently exhibit one or more processes. At the electron-microscopic level, the cytoplasm of positive cells contains few organelles, variously sized empty vacuoles, and a few electron-dense lysosome-like structures. Pinealocytes with synaptic ribbons have been found adjacent to immunoreactive cells. Double-immunoperoxidase staining for MRC OX6, MRC OX42, and ED1 results in OX6(-)/ED1(+)/OX42(+), OX6(-)/ED1(-)/OX42(+), and OX6(+)/ED1(-)/OX42(- )cells. These findings suggest that OX6-positive cells in the pineal can be considered as peripheral dendritic cells. The number of cells expressing MHC class II (Ia) antigen significantly increases in the pineal gland of rats after treatment with carbon tetrachloride (P<0.005). Our results indicate that at least some of the OX6-positive cells migrate into the gland from the circulation under these conditions.

Animals↗

Tryptophan hydroxylase mRNAs analysis by RT-PCR: preliminary report on the effect of noradrenaline in the neonatal rat pineal gland.

The levels of mRNAs coding for tryptophan hydroxylase (TPOH), the first enzyme in melatonin synthesis, have been investigated by quantitative reverse transcription of RNA, followed by polymerase chain reaction (RT-PCR), after stimulation of neonatal pineal organ cultures with Noradrenaline (NA). TPOH mRNAs were specifically amplified from various adult tissues, namely the pineal gland, raphe, retina, and kidney, but not the lung. PCR signals for TPOH were detected in the neonatal pineal gland in the absence of stimulation. Stimulation of neonatal pineal organ culture with 0.1 microM NA resulted in a significant increase (x2.5) in expression of TPOH mRNAs, whereas higher doses (1 and 10 microM) had no effect. All concentrations of NA enhanced melatonin secretion. Our results suggest that the level of TPOH mRNAs can be controlled by NA and that this effect might be implicated in the gene level regulation of the daily enzyme rhythm in the rat pineal gland.

Animals↗

Direct projections to the rat pineal gland via the stria medullaris thalami. An anterograde tracing study by use of horseradish peroxidase.

The possible presence of a direct nervous projection from the paraventricular nucleus (PVN) of the hypothalamus to the pineal gland of the rat was investigated by means of the anterograde neuron-tracing method using horseradish peroxidase. The tracer was injected unilaterally into the PVN and the animals were allowed to survive between 12 and 26 h. Numerous peroxidase-positive fibers were observed, ipsilateral to the injection site, in the stria medullaris thalami and could be followed into the medial habenular nucleus and the habenular commissure. From there, fibers penetrated into the deep pineal gland (lamina intercalaris), and further into the pineal stalk. These data support results of previous investigations describing retrograde labeling of the PVN following intrapineal injections of horseradish peroxidase and are in accordance with recent experiments demonstrating an influence of the PVN on electrical and biochemical activity of the pineal gland.

Afferent Pathways↗

Daily time course of the contents in monoamines and their metabolites in the pineal gland of Syrian hamster.

Twenty-four h changes in endogenous tyrosine hydroxylase (TH) activity, monoamines and their metabolites as well as N-acetyltransferase (NAT) activity and melatonin content were investigated in the Syrian hamster pineal gland. Both NAT activity and melatonin content exhibited an expected rise 8 h (0400 h) after darkness onset. Pineal dopamine (DA) and 3,4-dihydroxyphenyl acetic acid (DOPAC) levels were highest during the dark phase reaching a peak at 0200 h, just prior to the rise in melatonin production. However no significant difference were found in both norepinephrine (NE) and TH activity levels during the 24 h period. However, in the present study, the rat pineal gland exhibited a clear day/night difference in its TH activity. These data suggest a possible role of pineal DA in the induction of melatonin synthesis in addition to its function as a NE precursor in the Syrian hamster pineal gland and also indicate that catecholamines may act on the hamster pineal in a different manner than in the rat.

3,4-Dihydroxyphenylacetic Acid↗

Innervation of the cat pineal gland by neuropeptide Y-immunoreactive nerve fibers: an experimental immunohistochemical study.

An immunohistochemical study of the cat pineal gland was performed using a rabbit polyclonal antibody directed against neuropeptide Y (NPY) and an antibody directed against the C-terminal flanking peptide of neuropeptide Y (CPON). Numerous NPY- and CPON-immunoreactive (IR) nerve fibers were demonstrated throughout the gland and in the pineal capsule. The number of IR nerve fibers in the capsule was high and from this location fibers were observed to penetrate into the gland proper via the pineal connective tissue septa, often following the blood vessels. From the connective tissue septa IR fibers intruded into the parenchyma between the pinealocytes. Many IR nerve fibers were observed in the pineal stalk and in the habenular as well as the posterior commissural areas. The number of NPY/CPON-IR nerve fibers in pineal glands from animals bilaterally ganglionectomized two weeks before sacrifice was low. The source of most of the extrasympathetic NPY/CPONergic nerve fibers is probably the brain from where they enter the pineal via the pineal stalk. However, an origin of some of the fibers from parasympathetic ganglia cannot be excluded due to the presence of a few IR fibers in the pineal capsule of ganglionectomized animals. It is concluded that the cat pineal is richly innervated with NPYergic nerve fibers mostly of sympathetic origin. The posttranslational processing of the NPY promolecule results in the presence of both NPY and CPON in intrapineal nerve fibers.

Animals↗

Effect of isoprenaline on the output of melatonin from fetal and newborn lamb pineal glands in vitro.

We have measured the output of melatonin in vitro from the pineal glands of lambs in late gestation (135-143 days) and the early newborn period (4-8 days). The basal output of melatonin/mg pineal tissue was significantly lower in the fetal group (16.3 +/- 3.9 pg melatonin.(30 min)-1.mg-1) than in the newborn group (148.1 +/- 43.6 pg melatonin.(30 min)-1.mg-1). Addition of the beta-adrenergic agonist, isoprenaline (10(-5) mol/l) significantly stimulated melatonin output from both the fetal (to 24.9 +/- 3.9 pg melatonin.(30 min)-1.mg-1) and newborn pineals (to 247.7 +/- 67.3 pg melatonin.(30 min)-1.mg-1). Pineals from newborn lambs produced 9-10-fold more melatonin than the fetal pineal glands, before, during and after addition of isoprenaline. These results show that functional beta-adrenergic receptors are present before birth on the sheep pinealocyte and that there is a marked increase in the melatonin synthetic capacity of the pineal in the newborn period.

Adrenergic beta-Antagonists↗

Enhanced phosphoinositide hydrolysis in the pineal gland of spontaneously hypertensive rats.

We studied the phosphoinositide (PI) turnover in the pineal gland of spontaneously hypertensive rats (SHR) and compared it to that in Wistar-Kyoto and Wistar rats. Both basal and norepinephrine-stimulated PI turnover were significantly higher in the SHR than in the other strains. Prazosin (1 mumol/L) completely blocked the norepinephrine-induced PI hydrolysis in all groups. However, prazosin failed to lower the enhanced basal PI turnover in SHR. Pineal response to the cholinergic agonist carbachol was similar in all groups. These results suggest enhanced membrane phospholipid turnover in the pineal gland of SHR. Moreover, signaling via the alpha 1-adrenoceptor might be sensitized in the SHR.

Animals↗

Day-night rhythm of acetylcholine in the rat pineal gland.

Using high-performance-liquid-chromatography (HPLC) measurement of acetylcholine, choline acetyltransferase (ChAT) enzyme assay and anti-ChAT immunohistochemistry, we have investigated the expression of the cholinergic system in pineal glands of male rats. Glands procured during the day period (1200 h) contained significant amounts of acetylcholine (0.5 pmol/gland). A similar content was found in pineal glands after a 48 h culture period, i.e. when the intrapineal nerve fibres have degenerated. This strongly indicates that the pinealocytes are the cells which contain acetylcholine. To confirm this conclusion we demonstrate substantial ChAT-like immunoreactivity in pinealocytes. ChAT enzyme activity measured in homogenized glands (day period) was 7 +/- 3 nmol/mg per h. Acetylcholine content as well as ChAT enzyme activity increased about 10-fold in pineal glands during the night period (2400 h). The present study demonstrates for the first time the presence of a day-night rhythm of ChAT and acetylcholine in rat pinealocytes. The function of pineal acetylcholine is not clear, but there are indications that acetylcholine may depress noradrenaline release from intrapineal sympathetic fibres and hence melatonin synthesis.

Acetylcholine↗

Morphometric analysis of the synaptic ribbons and nerve vesicles of the cat pineal gland after electrical stimulation of the superior cervical ganglia.

The nerve vesicles and synaptic ribbons were quantified in the cat pineal gland after electrical stimulation of the pineal sympathetic nerve fibers. It has been shown that the bilateral electrical stimulation of the preganglionic fibers innervating the superior cervical ganglia (SCG) markedly reduces the number of dense cores of small dark vesicles (SDV) and, on the other hand, modifies the number and shape of the large dark vesicles (LDV). An increase in the number of the synaptic ribbons after stimulation of the SCG supports the hypothesis that the numerical reduction of dense cores of nerve vesicles in the cat pineal gland influences the level of synaptic ribbon formation.

Animals↗

Cell proliferation in the developing rat pineal gland. A bromodeoxyuridine immunohistochemical study.

The immunohistochemical detection of bromodeoxyuridine (BrdU) was used to study the cell proliferation in the developing rat pineal gland, from the appearance of pineal primordium in the embryonic day 15 (E15) until 30 days after birth. The results showed three different proliferative phases. From E15 to E21, the pineal gland shows a phase of rapid proliferation. The second phase corresponds to the first postnatal week, in which the number of labeled cells per surface unit decreases suddenly to values between 20% to 10% of those of embryonic period. From the second postnatal week onwards, the number of BrdU-positive cells progressively decreases.

Animals↗

Calcium influx through voltage-sensitive calcium channels regulates in vivo serotonin N-acetyltransferase (NAT) activity in hen retina and pineal gland.

The involvement of transmembrane transport of Ca2+ in regulation of retinal and pineal serotonin N-acetyltransferase (NAT) activity was studied in vivo in hens. Intramuscular administration to hens of organic antagonists of voltage-sensitive calcium channels (VSCC), nimodipine, nifedipine and verapamil, resulted in a marked inhibition of the nocturnal increase of NAT activity in retina and pineal gland. In both tissues the inhibitory effect of nimodipine on the night-time rise of NAT activity was abolished by Bay K 8644, a VSCC agonist. Bay K 8644 administered to hens at the end of the light phase of the light-dark cycle significantly enhanced the dark-dependent increase in retinal and pineal NAT activity. It is suggested that the nocturnal increase of NAT activity in hen retina and pineal gland is similarly regulated in vivo by Ca2+ influx through the VSCC.

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗

Rat pineal gland suppresses the injection stress-reactive ACTH outflow.

The involvement of the pineal gland in the regulation of the initial ACTH reactive outflow to short-term stress inducement was studied. The stress trial, consisting of the subcutaneous saline injection in the animal hind leg, elicited ACTH release, the peak appearing in the 5th min followed by a rapid downturn to the prestress level in the 15th min. The higher ACTH level in the pinealectomized rats was monitored in the 5th min. The results obtained show the implication of the pineal gland in restricting the magnitude of the initial stress-reactive ACTH outflow.

Adrenocorticotropic Hormone↗

Binding sites for melatonin in bovine pineal gland.

High-affinity binding of melatonin to crude membrane preparations of bovine pineal gland was examined by a rapid filtration procedure through Whatman GFB paper. Maximal melatonin binding was attained in 60 min at 37 degrees C, in 2 h at 25 degrees C and in 5 h at 0 degree C; at 25 and 37 degrees C it was 36 and 42% of that at 0 degree C. Specific binding was thermolabile and decreased following incubation with trypsin; it was also pH dependent, the maximum being observed at physiological pH. Melatonin binding was inhibited by the addition of monovalent or divalent ions to the incubation buffer. Subcellular fractionation studies indicated that 39 and 50% of binding was located in the pellets at 900 and 27,000 g whereas 11% was detected in the microsomal pellet. Scatchard analysis revealed a single population of binding sites with Kd = (7.0 +/- 1.5) 10(-7) M (mean +/- SEM, n = 4); binding site concentration ranged from 185 to 356 fmol/mg of protein. When various melatonin analogues were tested for their ability to inhibit (3H)-melatonin binding, the following Ki values (10(-7) M), were obtained: N-acetyl-serotonin 120, serotonin 130, 2-methyl indole 154, 5-hydroxytryptophol 218, 5-methoxytryptamine 266, 5-methoxytryptophol 660, tryptamine 1,740, 5-hydroxyindole acetic acid 3,455, 5-methoxyindole acetic acid 12,690, 5-hydroxytryptophan 13,600, and 6-hydroxymelatonin 55,550. Those results suggest that melatonin receptors may be present in the pineal gland.

Animals↗

[Circadian rhythm of ornithine decarboxylase and its endogenous high molecular weight inhibitor in rat pineal gland].

The biochemical mechanisms involved in circadian variations of the activity of ornithine decarboxylase (EC 4.1.1.17)--the rate-limiting enzyme of polyamine biosynthesis in rat pineal gland were studied. The enzyme was separated from its endogenous high molecular weight inhibitor by gel-filtration of the cytosol fraction from this organ through Sephadex G-100 in the presence of 250 mM NaCl. The inhibitor was similar in its molecular weight (30 000) and activity to ornithine decarboxylase inhibior from rat liver. The amount of the enzyme in the pineal gland undergoes much smaller circadian variations as compared to that of the inhibitor. It is concluded that the circadian variations of the ornithine decarboxylase activity in the pineal gland may be largely due to the changes in the enzyme/inhibitor ratio.

Animals↗

A stereological study of the effects of the pineal gland on the ventral prostate of the rat.

Several procedures known to influence pineal gland secretion were studied for their effects on the ventral prostate of rats. Stereological measurements were made of volume fractions of acini and glandular epithelium, surface fractions of glandular epithelium, and length fractions of acini. From these figures and the weights of the glands, the volumes, surface areas, and lengths of these features were calculated, as well as the mean epithelial heights, mean acinar diameters, and mean distances between glandular acini. None of these measurements differed in sham-operated controls, pinealectomized, or blinded rats. In blind-anosmic rats, however, there were significant decreases in the weights of the prostates and the volumes of the acini, reflecting significant decreases in the mean diameters of acini. None of the other parameters differed from those of controls, except the length fraction of acini. These results differ from those described by others following castration in rats, indicating that the action of the pineal gland on the ventral prostate may be different from that of androgen deprivation, in this species. Caution should therefore be exercised in the interpretation of the results of experiments in which prostate weights are used as indications of the actions of pineal hormones on the neuroendocrine-gonadal axis.

Animals↗