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PubMed1996

Nycthemeral expression of tryptophan hydroxylase mRNAs in the rat pineal gland.

The expression of mRNAs coding for tryptophan hydroxylase (TPOH), the first enzyme involved in melatonin synthesis, has been studied in the rat adult pineal gland at four different circadian time points by in-situ hybridization using an oligonucleotide probe. TPOH mRNAs were detected at all chosen time-points. The lowest level was seen at 13:00 h. Quantification of the hybridization signals demonstrated a significant increase in expression (+16.5%) between 13:00 h and 19:30 h, with the values at 23:30 h and 09:00 h being similar to that of 19:30 h. These results coincide with the nycthemeral rhythm of TPOH activity. Day/night rhythms in the rat pineal gland are controlled by noradrenaline release at lights-off, which results in an increase in cAMP levels and in gene activation. This study suggests that the TPOH gene might be one such gene whose activation is stimulated in this way.

Animals↗
PubMed2006

Relationship between nocturnal serotonin surge and melatonin onset in rodent pineal gland.

BACKGROUND: We have recently reported dynamic circadian rhythms of serotonin (5-HT, 5-hydroxytryptamine) output in the pineal gland of rat, which precedes the onset of N-acetylserotonin (NAS) and melatonin secretion at night. The present study was aimed at investigating in detail the relationship between 5-HT onset (5HT-on) and melatonin onset (MT-on) in multiple strains of rats and comparing them with those of hamsters. METHODS: Animals were maintained in chambers equipped with light (250 lux at cage levels) and ventilation in a temperature-controlled room. Following surgical implantation of a microdialysis probe in the pineal gland, animals were individually housed for on-line pineal microdialysis and for automated HPLC analysis of 5-HT and melatonin. Animals were under a light-dark cycle of 12:12 h for the duration of the experiments. RESULTS: All animals displayed dynamic 5-HT and melatonin rhythms at night. In all cases, 5HT-on (taken at 80% of the daily maximum levels) preceded MT-on (taken at 20% of the daily maximum levels). Within the same animals, 5HT-on as well as MT-on across multiple circadian cycles exhibited minimum variations under entrained conditions. Large inter-individual variations of both 5HT-on and MT-on were found in outbred rats and hamsters under entrained conditions. In comparison, inbred rats displayed very small individual variations of 5HT-on and MT-on. Importantly, we have uncovered a species-specific relationship of 5HT-on and MT-on. 5HT-on of rats, regardless of the strain, preceded MT-on of the same rats by 50 min. In contrast, 5HT-on of hamsters led MT-on by as much as 240 min. Thus, while a constant relationship of 5HT-on and MT-on exists for animals of the same species, the relative timings of 5HT-on and MT-on differ between animals of different species. CONCLUSION: These results suggest that both 5-HT and melatonin could serve as reliable markers of the circadian clock because of their day-to-day precision of onset timings within the same animals or within individuals of the same strain or same species. The results also demonstrate that data for MT-on cannot be compared directly between different species, and that 5HT-on may be a more reliable circadian marker when data from animals of different species are compared.

Journal Article↗
PubMed1991

Expression of S-antigen in retina, pineal gland, lens, and brain is directed by 5'-flanking sequences.

S-antigen (S-Ag) is an abundant protein of the retina and pineal gland that elicits experimental autoimmune uveitis and pinealocytis in several animal species. To study the elements regulating the expression of S-Ag, we generated transgenic mice expressing the chloramphenicol acetyl transferase (CAT) gene under the control of a 1.3-kilobase pair 5'-flanking segment of the mouse S-Ag gene. While all of the transgenic mice expressed CAT activity in the retina, in some animals CAT activity was also detected in the pineal gland, lens, and brain. Immunoblotting, polymerase chain reaction-mediated detection of RNA, and immunocyto-staining of transgenic tissues with antibodies to CAT and S-Ag established that the profile of expression of the transgene corresponded to that of S-Ag; both proteins were detectable in retinal photoreceptor cells, pinealocytes, lens fiber and epithelial cells, the cerebellum, and the cerebral cortex. These results indicate that S-Ag is expressed in a wider spectrum of the cell types than previously recognized and that a 1.3-kilobase pair S-Ag promoter segment contains sufficient information to direct appropriate tissue-specific gene expression in transgenic mice.

Animals↗
PubMed1999

Embryonic development of pineal gland vesicles: a morphological and morphometrical study in chick embryos.

Pineal cell aggregates in 5, 10 and 15 day-old chick embryos have been studied. Cell aggregates were classified into rosettes or vesicles and spheroid and ellipsoid vesicles distinguished. The number of pineal vesicles per unit of surface (vesicle density) was determined in three pineal portions: apical, anterior and posterior. By day 5, only cellular rosettes were found, mainly in the apical portion. After 10 and 15 days, the presence of rosettes was occasional. The posterior wall showed only small spheroid vesicles, while in the apical and anterior areas ellipsoid vesicles were also observed. Moreover, the spheroid/ellipsoid vesicle ratio increased from the 10th to the 15th day of incubation. The vesicle density decreased between the 10th and 15th day because of the increase in both vesicle and pineal size, without changes in the total number of vesicles. The results suggest that changes in vesicle morphology and density could be related to the functional activity of the pineal gland during embryonic development.

Animals↗
PubMed2003

Diurnal variation in p42/44 mitogen-activated protein kinase in the rat pineal gland.

In this study, we investigated whether there was a diurnal difference in mitogen-activated protein kinase (p42/44(MAPK)) phosphorylation in the rat pineal gland. Under a lighting regimen with 12h of darkness, there was a two- to four-fold increase in phosphorylated levels of MAPK kinase 1/2 (MEK1/2) and p42/44(MAPK) 2h after onset of darkness, an increase that was sustained for 8h. The increases in phosphorylated levels of MEK1/2 and p42/44(MAPK) occurred without increases in MEK1/2 and p42/44(MAPK) proteins. When rats were treated with propranolol 1h before onset of darkness or subjected to continuous light exposure during the dark phase, the nocturnal increase in MEK1/2 and p42/44(MAPK) phosphorylation was reduced. Acute light exposure during darkness caused a decline in MEK1/2 and p42/44(MAPK) phosphorylation within 30 min of light exposure. These results indicate the presence of a diurnal difference in MEK1/2 and p42/44(MAPK) phosphorylation in the rat pineal gland that is under adrenergic control.

Animals↗
PubMed1988

Purification and characterization of DSIP-like material from ovine pineal glands: possible peptide-protein interaction.

The nonapeptide delta-sleep-inducing peptide (DSIP) has been isolated from venous blood of rabbits induced to sleep. Numerous reports have described sleep as well as extra-sleep effects. Radiochemical and immunochemical data suggest a relationship of DSIP with the pineal gland supported by interactions of this peptide with pineal functions such as the serotonin N-acetyltransferase activity. In order to demonstrate the natural occurrence of DSIP-like material associated with high Mr proteins in the ovine pineal, organs were water-extracted and fractionated by ultrafiltration and gel filtration. Radioimmunoassay (RIA) for DSIP-like fragments of the fractions revealed considerable amounts of pineal DSIP-like immunoreactivity (DSIP-LI) apparently existing in small as well as large molecular forms. Acidification of large DSIP-LI forms resulted in the elution from Sephadex G-50 of Mr less than or equal to 1,000 DSIP-like material. This free DSIP-LI form coeluted with the synthetic DSIP nonapeptide from microBondapak C18 on high-performance liquid chromatography. The results, therefore, appear to indicate the presence of a (biospecific) noncovalent intermolecular interaction of DSIP (1-9) with proteins (Mr greater than or equal to 10,000) of the ovine pineal gland.

Animals↗
PubMed1996

Presence of nitric oxide synthase in the sheep pineal gland: an experimental immunohistochemical study.

By use of immunohistochemistry, a dense network of nerve fibres immunoreactive to the neuronal form of nitric oxide synthase (NOS, subtype I) was demonstrated in the pineal gland of sheep. The NOS-immunoreactive fibres were located in the pineal capsule and the connective tissue septae of the gland, but fibres were also present intraparenchymally between the pinealocytes. NOS-immunoreactive nerve fibres were still present in the gland 1 month after bilateral removal of the superior cervical ganglia. By use of an antibody directed against endothelial NOS (subtype III), only pineal blood vessels were stained. This staining was still present in the ganglionectomized animals. No difference was found in the staining between the control animals and the ganglionectomized ones. The pinealocytes were not stained, neither by the antibody against neuronal NOS nor by the antibody against endothelial NOS. By use of double immunohistochemical stainings, NOS was in many nerve fibres colocalized with vasoactive intestinal peptide. Western blot analysis of supernatant fractions of sheep pineal homogenates showed the presence of a band corresponding to the neuronal NOS. Thus, the present data show a prominent innervation of the sheep pineal gland with NOS-immunoreactive nerve fibres with their origin outside the sympathetic nervous system, indicating an influence of NO on the pinealocyte metabolism from non-sympathetic nerve fibres in this species. The presence of NOS in both perivascular nerve fibres and the endothelium of the blood vessels of the gland suggests a role of NO in the regulation of the circulation of the sheep pineal gland.

Animals↗
PubMedDate not supplied

Unique pineal gland metastasis of clear cell renal carcinoma: case report and review of the literature.

The metastatic involvement of the pineal gland is an extremely unusual event; it has a 4% incidence in patients with disseminated neoplasias. Most metastatic pineal lesions are asymptomatic. Only in a small number of cases the symptoms produced by metastatic involvement of this organ precede those of the primary tumor or those of another metastatic site. To our knowledge the herein reported case is the first in which the pineal gland was apparently the unique metastatic site of a primitive kidney carcinoma and where the symptoms produced by metastasis in the pineal region were the first sign of the disease.

Aged↗
PubMed1997

Peptidergic cells in the mammalian pineal gland. Morphological indications for a paracrine regulation of the pinealocyte.

Several neuropeptides are present in the mammalian pineal gland. Most of these peptides, e.g. neuropeptide Y, vasoactive intestinal peptide, and peptide histidine isoleucine, are located in nerve fibres innervating the gland. In some mammalian species, neuropeptides are also found in cells scattered in the pineal parenchyma. In the rat, bipolar cells immunoreactive for somatostatin are present, just as cells containing mRNA encoding somatostatin can be detected in the gland by in situ hybridisation. In the pineal gland of the European hamster, many cells are immunoreactive for enkephalin. Ultrastructural cytochemical analysis of these cells reveals a pinealocyte morphology. Processes from the opioidergic pinealocytes terminate in the parenchyma between the non-immunoreactive pinealocytes. Some of the processes contain small clear and large dense core vesicles and end in club shaped swellings which make synapse-like contacts with other pinealocytes. The ultrastructural morphology suggests that the opioidergic cells exert a paracrine regulation on other pinealocytes.

Animals↗
PubMed1975

Ultrastructural study of the pineal gland of the chicken (Gallus gallus).

The authors studied the pineal glands of chickens (Gallus gallus) between the ages of 2 and 5 days with the electron microscope. They described two distinct areas in the parenchyma of the organ: the follicles, formed by two cellular categories, the pineal A and B cells, and the parafollicular zone, which surrounds and separates the follicles from the connective walls. This area is formed, fundamentally, by two types of cells and nerve fibers. These cells occasionally delimit cavities. We propound the hypothesis of the possible transformation of the parafollicular zone into follicles.

Age Factors↗
PubMed1997

Pinealocyte ultrastructure in mutant mice that lack sympathetic innervation to the pineal gland.

Pinealocyte synaptic ribbons (SR) and dense-cored vesicles (DCV) were quantitated in mice that have developmental deficiencies in the sympathetic innervation of the pineal gland due to a null mutation for the p75 low-affinity NGF receptor (NGFR). SR exhibited a significant increase whereas there was a significant decline in the frequency of DCV in mutant mice. These findings support the hypothesis that pinealocyte SR and DCV are regulated by neural mechanisms associated with sympathetic system. Ultrastructural characteristics of pinealocytes in mutants included degenerative changes that culminated in the breakdown of cellular components and the accumulation of flocculent-containing vesicles within the cytoplasm. Ribosomal lamellar complexes were also commonly seen in pinealocytes of mutant rats. Although the mutant pineal gland exhibited signs of metabolic imbalances, the cytoarchitecture of the gland (e.g., vascular compartment) and differentiation of the cells were generally unaffected by developmental deficiencies in the gland's innervation.

Animals↗
PubMed2003

Influences of a light-dark profile and the pineal gland on the hypnotic activity of melatonin in mice and rats.

We have investigated the influences of the light-dark cycle and the pineal gland on the hypnotic activity of melatonin in rats and mice. The results showed that melatonin significantly shortened time to sleep onset and wakefulness time, increased slow wave sleep, paradoxical sleep, and total sleep time in rats during the light phase of a 12-h light:12-h dark cycle, by electroencephalogram recording. However, during the dark phase it had almost no significant sleep-promoting effect except shortened time to sleep onset. Melatonin exhibited more potent sleep-promoting effect in rats exposed to constant light compared with rats exposed to 12:12-h light:dark at 2000 h. Melatonin markedly prolonged sleeping time in the mice exposed to constant illumination. It was found that pinealectomy was an important factor that influenced the hypnotic activity of melatonin. When melatonin was administered to pinealectomized mice, the hypnotic activity of melatonin was more intense compared with sham-operated mice. These results demonstrated that the hypnotic activity of melatonin displayed a light-dependence manner. These results suggested that light exposure and the functional state of the pineal gland could substantially impact the hypnotic activity of melatonin at pharmacological dosage.

Administration, Oral↗
PubMed1990

Synaptic ribbons of the rat pineal gland: responses to in-vivo and in-vitro treatment with inhibitors of protein synthesis.

To elucidate the role of protein synthesis in the nocturnal increase of synaptic ribbons in the rat pineal gland, actinomycin-D, which inhibits transcription, and cycloheximide, an inhibitor of translation, were used. To assure that the drugs were effective and to relate morphological changes to pineal biosynthetic phenomena, the activity of N-acetyltransferase and levels of pineal indoleamine were measured. Results of in-vivo, short-term and long-term treatment with either drug suggest that transcription of proteins related to synaptic ribbon formation occurs during the first half of the light phase, whereas translation takes place during the first few hours of the dark phase. In contrast, proteins involved in enhanced melatonin synthesis are transcribed and translated during the first few hours of the dark phase. In vitro, preincubation with inhibitors of protein synthesis abolished the increase in the numbers of synaptic ribbons after stimulation with dibutyryl-adenosine-cyclic-monophosphate, indicating that the results of the in-vivo experiments are due to an interaction of the drugs with the pineal gland itself. The present study shows that, although in the rat pineal enhanced melatonin synthesis and increased numbers of synaptic ribbons occur at the same time, transcription of proteins involved in both rhythms is temporally separated.

Acetyltransferases↗
PubMed1984

Analysis of melatonin, 5-methoxytryptophol and 5-methoxyindoleacetic acid in the pineal gland and retina of hamster by capillary column gas chromatography-mass spectrometry.

A specific capillary column gas chromatographic-mass spectrometric method was used to determine 5-methoxyindoles in the pineal gland and retina of the golden hamster during a light-dark (14:10) cycle. In the pineal gland, the mean levels of melatonin ranged from 0.15 to 2.4 pmol per gland, with a maximum in the dark. The levels of 5-methoxytryptophol and 5-methoxyindoleacetic acid were in the same range, but peaked during light. In the retina the levels of melatonin were about 100 pmol/g, and seemed not to differ between light and dark. The level of 5-methoxyindoleacetic acid were in the same range during light but were below the detection limit during dark.

Animals↗
PubMed1981

Gap junctions between pinealocytes. A freeze-fracture study of the pineal gland in rats.

The intercellular junctions between the pinealocytes of male rats were investigated by freeze-fracture and conventional electron microscopy. Our findings reveal that the intercellular contacts between pineal cells, formerly described as zonulae adhaerentes or zonulae occludentes, are in fact gap junctions which are difficult to characterize in thin sections due to their peculiar geometrical arrangement, which is in the form of "fenestrated" communicating zonules. The arrangement of these communicating zonules around rudimentary lumina of pineal clusters and rare transitions between tight and gap junctions may point to phylogenetic transformations of occluding into communicating zonules, corresponding with the change of the pineal gland from a sensory to a secretory organ. Alternatively, these tight-to-gap junctional transitions may reflect the periodic (circadian or seasonal) activity of the pineal gland.

Animals↗
PubMed1975

Effect of gonadotropic hormones on the rat pineal gland A fluorescence histochemical and biochemical study.

Changes in the number of autofluorescent and serotonin-containing pinealocytes have been studied after gonadotropic hormone administration using fluorescence histochemistry. Moreover, the 5-HT levels in the pineal gland were determined biochemically under the same experimental conditions. The results suggests that pineal 5-HT levels are not influenced by increased gonadotropin levels, while the amount of autofluorescent pinealocytes increases after gonadotropin injections. It has been postulated that differences in androgenic hormone levels could not cause the changes in the number of autofluorescent pinealocytes. The reaction of the pineal gland to increased gonadotropin levels seemed important in view of the regulatory function of the epiphysis on the hypothalamo-hypophyseo-gonadal axis.

Animals↗
PubMed1997

Immunocytochemical identification of pinopsin in pineal glands of chicken and pigeon.

Pinopsin is a blue-sensitive photoreceptive molecule possibly involved in photic entrainment of the circadian pacemaker in the chicken pineal gland. To characterize pinopsin as a circadian photoreceptor, antibodies were raised against the C-terminal portion of pinopsin. As expected from the divergence of the amino acid sequence of this region, the resultant antibody cross-reacted with neither chicken rhodopsin nor red-sensitive cone pigment (chicken red). In Western blot analysis, the antibody stained a single band of 42-kDa protein in a detergent-extract of chicken pineal membranes, suggesting that pinopsin (calculated molecular weight, 38187) might be glycosylated and/or palmitoylated. Immunocytochemical examination of pineal sections of the chicken and the pigeon with this antibody revealed strong positive images for most of the membrane structures in the lumen of the follicles. This antibody also stained string- and bulb-shaped structures of the chicken parafollicular cells, the morphology of which resembles those of retinal photoreceptor cells. In contrast to the predominant distribution of pinopsin, a monoclonal antibody specific for chicken red stained a smaller number of membrane structures in the lumen of chicken pineal follicles. These results strongly suggest that the chicken pineal gland contains at least two types of photoreceptive molecules, pinopsin (major) and chicken red (minor). We show that the former molecule is localized in parafollicular pinealocytes and in the outer segments of pinealocytes that make contact with the follicular lumen.

Amino Acid Sequence↗
PubMed1982

Darkness-elicited changes in rat reproduction not mediated by pineal gland.

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.

Animals↗