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PubMed2005

The rhythm and blues of gene expression in the rodent pineal gland.

In all vertebrates, melatonin is rhythmically synthesized in the pineal gland and functions as a hormonal message, encoding for the duration of night. In rodents, the nocturnal rise and fall of the arylalkylamine N-ace-tyltransferase (AA-NAT) activity controls the rhythmic synthesis of melatonin. This rhythm is centered around the transcriptional regulation of the AA-NAT by two norepinephrine-inducible transcription factors, the activator CREB (Ca2+/cAMP-response element binding protein) and the inhibitor ICER (inducible cAMP early repressor). CREB is activated by phosphorylation, which is one of the fastest responses in pinealocytes upon adrenergic stimulation, occurring within minutes. ICER in turn accumulates only after several hours, a time gap resulting from the required de novo protein synthesis upon adrenergic stimulation. However, these molecular components of neuroendocrine signaling in the rodent pineal gland are supplemented by the impact of a variety of neurotransmitters and neuromodulators, and by translational and post-translational mechanisms. By molecular crosstalk, those different inputs on pinealocytes seem to fine-tune the shape of the melatonin signal, by interacting at various levels with the NE/cAMP/pCREB/ICER pathway. In addition, these alternate signaling routes may be important in acute "emergency" situations. Together, concerted signaling events in the rodent pineal gland help to generate a stable and reliable hormonal message of darkness for the body, that, however, can be altered rapidly upon sudden and unexpected "error" signals.

Animals↗

PubMed1998

Neuronal projections from the mesencephalic raphe nuclear complex to the suprachiasmatic nucleus and the deep pineal gland of the golden hamster (Mesocricetus auratus).

Neuronal projections from the mesencephalic raphe system to the suprachiasmatic nucleus and the pineal complex were mapped in this study of the golden hamster, by use of the anterograde tracer Phaseolus vulgaris-leucoagglutinin and the retrograde tracer cholera toxin subunit B. From the median raphe nucleus, a rostral projection ascended in the ventral part of the mesencephalon to continue in the medial forebrain bundle of the forebrain. Nerve fibres from this bundle innervated the ventral and medial parts of the suprachiasmatic nucleus. At the level of the interpeduncular nucleus of the mesencephalon, fibres of the ventral bundle bent dorsally to reach the epithalamic area and to continue in the forebrain in a periventricular position. Some of these fibres innervated the dorsal tip of the suprachiasmatic nucleus. The dorsal raphe nucleus was the origin of a nerve fibre bundle, located in the periaqueductal gray of the mesencephalon, innervating the deep pineal gland and pineal stalk. Injection of cholera toxin B into the suprachiasmatic nucleus labelled cells in the median raphe. Combination of the retrograde tracing from the suprachiasmatic nucleus and serotonin transmitter immunohistochemistry showed that some of the cholera toxin B-immunoreactive nerve cells also contained serotonin. Thus, this study of the golden hamster shows a serotonergic projection from the median raphe nucleus to the suprachiasmatic nucleus and a projection from the dorsal raphe nucleus to the deep pineal gland supporting physiological indications of an influence of serotonin on the photoreceptive circadian system of the brain.

Animals↗

PubMedDate not supplied

Development of the mammalian pineal gland.

The literature concerning the morphological, biochemical, and pharmacological development of the mammalian pineal gland is reviewed with an emphasis on the ontogeny of pineal innervation by noradrenergic neurons. Mature pineal responses to various pharmacological manipulations and the well-characterized rhythms in pineal biochemistry appear only after postnatal axogenesis of sympathetic neurons, synaptogenesis between sympathetic nerve terminals and pineal adrenergic receptors, and pineal nerve terminal synthesis, storage, and inactivation of catecholamine neurotransmitter. Since postsynaptic biochemical capacities for pineal enzymatic and endocrine function appear at a relatively early age, it is proposed that development of presynaptic inputs to the pineal is rate-limiting in the maturation of the pineal gland as a circadian neuroendocrine transducer. Because of this, the pineal gland is an excellent model for the functional development of noradrenergic inputs to selective target organs.

Acetylserotonin O-Methyltransferase↗

PubMed1990

Choline acetyltransferase in bovine pineal gland.

Recent studies from our laboratories have shown that the bovine pineal gland contains a muscarinic cholinergic receptor with a Kd value of 0.423 +/- 0.010 nM and a Bmax value of 69.75 +/- 20.91 fmol/mg protein. In order to substantiate further the possible existence of a pineal cholinergic transmission, we have measured the activity of choline acetyltransferase and delineated its kinetic properties in the bovine pineal gland. This enzyme exhibited an activity of 0.0339 +/- 0.0042 nmol/mg protein/min. Furthermore, the bovine pineal choline acetyltransferase possessed a Km value of 124.86 +/- 24.06 microM and a Vmax value of 0.0598 +/- 0.0034 nmol/mg protein/min for acetyl CoA, and a Km value of 3.11 +/- 0.94 mM and a Vmax value of 0.0155 +/- 0.0016 nmol/mg protein/min for choline. The presence of muscarinic cholinergic receptors along with a specific choline acetyltransferase are supportive evidences that the bovine pineal gland may receive cholinergic innervation.

Animals↗

PubMed2002

Morphometric parameters of the human pineal gland in relation to age, body weight and height.

The aim of the study was to compare the size, weight, volume and density of the pineal gland in several groups divided by age, body weight and height. 80 human pineal glands were included in the study. Obtained data were statistically analysed by means of Statistica by Statsoft to check existing differences. Obtained data show some significant differences between pineal gland morphometry in weight- and height-related groups. However, these differences do not influence pineal volume significantly. Differences between the pineal gland volume in the maximal and in the minimal weight groups may suggest some relationship between the gland's structure and body weight. No age-related changes in the morphometry of the pineal gland were observed.

Adolescent↗

PubMed1994

Cytochemical localization of monoamine oxidase in the pig pineal gland.

Monoamine oxidase plays an important role in the regulation of monoamine levels in the pineal gland by oxidative deamination of serotonin to 5-hydroxyindole acetic acid. The ultrastructural sites of this process are still not well known. In the present study the pig pineal glands were examined by using two cytochemical methods for demonstration of MAO activity at the ultrastructural level. The use of selective inhibitors of MAO-A (clorgyline and amiflamine) and MAO-B (deprenyl) showed that MAO-A was localized predominantly in noradrenergic nerve terminals as well as in the cell membrane of endothelial cells. MAO-B activity was localized in pinealocytes. However, in the outer membranes of cytoplasmic dense bodies, which are characteristic cytoplasmic structures of pig pinealocyte, we noticed the activity of both MAO-A and MAO-B enzymes. The MAO-A was localized exclusively in dense bodies within bulbous ending of pinealocyte processes present in the perivascular space. Our results show that in pig pineal gland there are two forms of MAO localized in different compartments of the pineal tissue. The most important result of our study is the presence of MAO in cytoplasmic dense bodies, which points to these bodies as to the sites of oxidative deamination and confirms their participation in the secretory process in pig pineal gland.

Animals↗

PubMed1995

The pineal gland: a comparative MR imaging study in children and adults with respect to normal anatomical variations and pineal cysts.

This study was undertaken to evaluate the variations in appearance of the normal pineal gland. The findings of 1000 consecutive MR imaging examinations obtained at 0.5 T were studied. The age of the patients ranged from 1 day to 83 years, and findings in children and adults were compared. In all age groups the pineal gland appeared mainly in three forms: (1) nodule-like, (2) crescent-like and (3) ring-like. Overall prevalences of these forms were 52%, 26% and 22%, respectively. Apparent differences in frequencies were evident in children and adults with respect to the crescent- and ring-like types. Cystiform pineal lesions 5 mm or larger in one diameter (anteroposterior, sagittal or transverse) were taken to be true pineal cysts, when compared with the gland's ring-like appearance (less than 5 mm). Pineal cysts had a prevalence of 0.6% in children and 2.6% in adults. No symptomatic pineal cyst with mass effect on the lamina tecti was detected in the series. Besides identifying the three anatomical types of the pineal gland as seen on MR imaging and addressing the potential significance of differences in their frequencies in children and adults, the author tries to explain the previous discrepancy between the MR imaging and autopsy series findings with respect to frequencies of the pineal cysts.

Adolescent↗

PubMed1991

Neuropeptide Y: an endogenous inhibitor of norepinephrine-stimulated melatonin secretion in the rat pineal gland.

The biosynthesis of the hormone melatonin (MEL) by the mammalian pineal gland has been thought to be regulated strictly by stimulatory factors, most predominantly norepinephrine (NE), released from the sympathetic nerve fibers which heavily innervate the gland. Evidence from many investigators suggests that sympathetic fibers may colocalize other neuroactive factors in addition to NE. One of these factors is neuropeptide Y (NPY), which has been found in the nerve fibers of the pineal gland. The present study sought to explore potential interactions between NE and NPY in the regulation of pineal MEL secretion. Specific, saturable, and reversible binding of 125I-NPY to intact cultured pinealocytes was measured with an affinity constant of 1 nM and an NPY binding site density of 0.04 pmol/mg of protein. In addition, cell culture studies revealed that NPY represents a potent (IC50 of 0.4 nM) endogenous inhibitor of NE-stimulated MEL secretion. However, this inhibition is accompanied by only a modest reduction (35%) of cyclic AMP accumulation. These findings reinforce the view that the mammalian pineal gland, which appears to integrate both inhibitory as well as stimulatory signals, is an important model of autonomic function, particularly in the context of biological rhythmicity.

Animals↗

PubMed2006

Evolution of the vertebrate pineal gland: the AANAT hypothesis.

The defining feature of the pineal gland is the capacity to function as a melatonin factory that operates on a approximately 24 h schedule, reflecting the unique synthetic capacities of the pinealocyte. Melatonin synthesis is typically elevated at night and serves to provide the organism with a signal of nighttime. Melatonin levels can be viewed as hands of the clock. Issues relating to the evolutionary events leading up to the immergence of this system have not received significant attention. When did melatonin synthesis appear in the evolutionary line leading to vertebrates? When did a distinct pineal gland first appear? What were the forces driving this evolutionary trend? As more knowledge has grown about the pinealocyte and the relationship it has to retinal photoreceptors, it has become possible to generate a plausible hypothesis to explain how the pineal gland and the melatonin rhythm evolved. At the heart of the hypothesis is the melatonin rhythm enzyme arylalkylamine N-acetyltransferase (AANAT). The advances supporting the hypothesis will be reviewed here and expanded beyond the original foundation; the hypothesis and its implications will be addressed.

Animals↗

PubMed2005

Morphological and biochemical study of the pineal gland of pregnant and non-pregnant female vizcachas (Lagostomus maximus maximus).

OBJECTIVES: The morphological and biochemical aspects of the pineal gland of pregnant and non-pregnant female vizcachas were investigated to examine whether there is a relationship between the pregnancy and this gland. METHODS: Pregnant and non-pregnant adult female vizcachas were used. The ultrastructure of type I cell (dark and light pinealocytes) was studied. Pineal and serum melatonin were investigated by radioimmunoassay. Radiometric methods were applied to investigate the arylalkylamine-N-acetyltransferase (AA-NAT) and hydroxyindole-O-methyltransferase (HIOMT) activities. RESULTS: The morphological study showed the existence of dark and light pinealocytes. The differences between both cells are slight. However, the appearance of dark pinealocytes suggests that they have a high activity. These cells were more abundant in the pineal gland of pregnant females. The pineal melatonin levels and the AA-NAT activity were higher in the pregnant females. Serum melatonin and HIOMT activity did not show significant differences. CONCLUSION: Our results suggest that the morphological and biochemical changes in the pineal gland of pregnant vizcachas are associated with the gestation. However, the relation between the pineal gland and the pregnancy is complex and further studies are necessaries to clearly establish it.

Acetylserotonin O-Methyltransferase↗

PubMed1982

Ultrastructure of the pineal gland of the eastern chipmunk (Tamias striatus).

The ultrastructure of the pineal gland of the wild-captured eastern chipmunk (Tamias striatus) was examined. A homogenous population of pinealocytes was the characteristic cellular element of the chipmunk pineal gland. Often, pinealocytes showed a folliclelike arrangement. Mitochondria, Golgi apparatus, granular endoplasmic reticulum, lysosomes, centrioles, dense-core vesicles, clear vesicles, glycogen particles, and microtubules were consistent components of the pinealocyte cytoplasm. The extraordinary ultrastructural feature of the chipmunk pinealocyte was the presence of extremely large numbers of "synaptic" ribbons. The number of "synaptic" ribbons in this species exceeded by a factor of five to 30 times that found in any species previously reported. In addition to pinealocytes, the pineal parenchyma contained glial cells (oligodendrocytes and fibrous astrocytes). Capillaries of the pineal gland of the chipmunk consisted of a fenestrated endothelium. Adrenergic nerve terminals were relatively sparse.

Animals↗

PubMed2002

Adrenergic inducibility of AP-1 binding in the rat pineal gland depends on prior photoperiod.

The main known function of the pineal gland in mammals is the temporal synchronization of physiological rhythms to seasonal changes of day length (photoperiod). In rat, the transcription factor activating protein-1 (AP-1) displays a circadian rhythm in its DNA binding in the pineal gland, which results from the rhythmic expression of Fra-2. We postulated that, if AP-1 is an important component of pineal gland functioning, then variations in photoperiodic conditions should lead to an adaptation of the AP-1 binding rhythm. Here we show that AP-1 binding patterns adapt to variations in lighting conditions, in the same way as the rhythm of arylalkylamine-N-acetyltransferase (AA-NAT) activity. This adaptation appeared to result from photoperiodic adaptation of the rhythmic fra-2 gene expression and was reflected by an adapted delay between the onset of night and the acrophase of the nocturnal peak. We further showed that photoperiodic adaptation of both the AP-1 binding and AA-NAT activity rhythms resulted from adapted changes in adrenergic inducibility of both variables at night onset. We finally provided evidence that AP-1 shared with the CREM gene encoding the transcriptional repressor protein inducible cAMP early repressor (ICER) the ability to be hypersensitive or subsensitive to adrenergic stimuli, depending on prior photoperiod.

Adaptation, Physiological↗

PubMed1981

Ultrastructural study of the embryonic development in the rat pineal gland.

The ultrastructure of the albino rat embryo pineal gland was studied from day 13 of development through birth. In the first stages (13-16.5 days of development) the pineal evagination presents a barely differentiated epithelium. From 17 days onward the transformation of the pineal gland from a tubular evagination into a compact organ occurs. The obliteration of the recess takes place by means of two mechanisms: (a) multiple foldings of the epithelium which determine an approximation and fusion of the walls of the recess, and (b) occupation of the lumen by cells extruded from the pineal epithelium. Embryos of 18-21 days of gestation still show remains of the pineal recess. From day 16.5 onward elements of the pineal parenchyma have been found outside the pineal epithelium contour. They contact with the mesenchymal cells without a basal lamina separating both elements. Day 20 marks the beginning of recognizable differentiation of pineal cellular types. However, in the newborn rat these types are not yet clearly established.

Animals↗

PubMed1984

In-vitro uptake and metabolism of [3H]-5-hydroxytryptamine in the pineal glands of the rabbit, rat and hamster. A comparative study with the use of autoradiography, chromatography and liquid-scintillation counting.

Pineal glands of rat, rabbit and hamster were incubated during day or night in Merlis' fluid containing [3H]-5-hydroxytryptamine (=[3H]-HT) by the use of a 20-min pulse with or without postincubation in "cold" medium for 15, 30, 45 or 60 min. (1) Selective autoradiographic labeling was observed in sympathetic nerve terminals; this reaction was missing after bilateral surgical removal of the superior cervical ganglia. In contrast, a scarce and diffuse labeling was found in pinealocytes (Pi) and interstitial cells (IC) of both untreated and ganglionectomized animals. (2) With the use of thin-layer chromatography, it could be shown in the rat that the well-known indoles of the pineal gland are formed from [3H]-HT. (3) During preparation for electron microscopy (EM), the total loss of indoles from pineal glands was studied by means of liquid-scintillation counting; approximately 57% of the radioactivity of the pineal glands was released into EM-processing solutions, mainly into the glutaraldehyde fixative. In summary, our results show that in this type of experiment with pineal glands of mammals, the routinely used EM-procedures are inadequate to visualize the uptake and metabolism of exogenous indoles in Pi and IC. Furthermore, the data differ considerably from those obtained with the pineal organs of several reptilian and avian species when a similar cytological procedure is used. It appears that protein(s) located in the densely packed vesicles of the pineal cells of sauropsids, homologous to mammalian pinealocytes, may play a crucial role in indole binding (specific indole-binding proteins); this may help to interpret the diverging results obtained in different amniotes.

Animals↗

PubMed2003

Specific distribution pattern of nerve fibers containing catecholamine-synthesizing enzymes, neuropeptide Y (NPY) and C-terminal flanking peptide of NPY (CPON) in the pineal gland of the chinchilla (Chinchilla laniger)--an immunohistochemical study.

The sympathetic nerve fibers originating from the superior cervical ganglia and supplying the pineal gland play the most important role in the control of the pineal activity in mammals. NPY and CPON are also present in the majority of the pinealopetal sympathetic neurons. In this study, immunohistochemical techniques were used to demonstrate the existence and coexistence of tyrosine hydroxylase (TH), dopamine beta-hydroxylase (DbetaH) as well as NPY and CPON in the nerve fibers supplying the chinchilla pineal gland. Ten two-year-old female chinchillas housed in natural light conditions were used in the study. The pineals were fixed by perfusion. ABC immunohistochemical technique and immunofluorescence labelling method were employed. TH-immunoreactive (TH-IR) varicose nerve fibers were observed in the pineal gland as well as in the posterior commissural area. Within the chinchilla pineal gland, TH-IR nerve fibers were located in the capsule and connective tissue septa. Numerous varicose TH-IR branches penetrated into the parenchyma and formed a network showing the highest density in the proximal region of the gland. In the central and distal parts of the pineal parenchyma, a subtle network, composed of thin varicose nerve branches, was observed. Double immunostaining revealed that the majority of TH-IR nerve fibers was positive for DbetaH or NPY. TH- and DbetaH-positive neuron-like cells were observed in the proximal region of the gland. The pattern of pineal innervation immunoreactive to CPON was similar to the innervation containing NPY, TH and DbetaH. The chinchilla intrapineal innervation containing TH, DbetaH, NPY and CPON is characterized by the higher density in the proximal part of the gland than in the middle and distal ones. The specific feature of the chinchilla pineal is also the presence of single TH/DbetaH-immunoreactive neuron-like cells in the proximal part of the gland.

Animals↗

PubMed1997

Expression and nocturnal increase of type II iodothyronine deiodinase mRNA in rat pineal gland.

It has been demonstrated that thyroxine deiodinating activity is present in rat pineal gland, and its activity increases significantly during the night time. We have studied whether mRNA for type II iodothyronine deiodinase is expressed in rat pineal gland and whether the nocturnal rise of pineal T4 deiodinating activity is due to the change in type II iodothyronine deiodinase mRNA level. Reverse transcription-polymerase chain reaction amplification and Northern blot analyses have demonstrated that type II iodothyronine deiodinase mRNA is expressed in rat pineal gland and its mRNA level increases markedly at midnight. These results suggest that the nocturnal rise in pineal T4 deiodinating activity is due to the change in type II iodothyronine deiodinase mRNA level.

Animals↗

PubMed1989

Histopathologic features of DMBA-induced mammary tumors in blind-underfed, blind-anosmic, and blind cold-exposed rats: influence of the pineal gland.

The influence of the pineal gland on the histopathologic changes induced in mammary glands by 7, 12-dimethylbenz(a)-anthracene (DMBA) has been studied. To achieve the maximum expression of pineal action in animals, 28-day-old female Sprague-Dawley rats were subjected to one of the following treatments: blindness + olfactory bulbectomy (BA), blindness + underfeeding (BU), or blindness + cold exposure (BC). Half of the rats in each group were also pinealectomized. Animals that were intact or only pinealectomized served as controls. Each animal received a single intragastrical dose of DMBA (20 mg) 4 weeks after performing the treatments mentioned above. At 28 weeks of age animals were sacrificed, and mammary tumors as well as mammary glands without macroscopic lesions were collected. Animals of the BU group failed to develop any tumors throughout the 20-week control period. In the remaining groups, tumors were found; the relation between number of adenocarcinomas (AC) and fibroadenomas was similar, always with a greater incidence of ACs. Some ACs showed areas with changes in the ductal epithelium that are considered signs of tumoral regression; pinealectomized animals had the lowest incidence of ACs with signs of regression. Ductule hyperplasia, considered as a premalignant lesion, was more frequent in pinealectomized rats than in controls and in animals with enhanced pineal function (BA, BU, and BC). Some nontumoral mammary glands showed dysplastic lesions; these lesions were less frequent in BU rats than in controls. We conclude that suppression of pineal function sensitizes the ductule cells to the initiation of neoplastic processes and hinders the development of regressive changes.

9,10-Dimethyl-1,2-benzanthracene↗

PubMed2001

Role of extracellular calcium on the regulation of melatonin synthesis in the Syrian hamster pineal gland.

The role of extracellular calcium on the induction of cyclic AMP production, N-acetyltransferase (NAT) activity and melatonin synthesis, induced by forskolin, was investigated in Syrian hamster pineal glands. Pineals were removed from hamsters killed either in the first half of the normal dark period or late in the dark period. Forskolin immediately increased NAT activity and melatonin levels only when the glands were collected late in the dark period, while in the first half of the dark phase, hamster pineals responded to forskolin with an increase of NAT activity and melatonin production only after 6 hr of incubation. The absence of calcium prevented the induction of melatonin synthesis by forskolin only when glands were collected early in the dark phase and incubated for 6 hr. In the second half of the normal dark period removal of calcium markedly decreased NAT activity and melatonin levels in glands incubated with forskolin for either 4 or 6 hr. However, the absence of extracellular calcium had no significant effect on the induction of cyclic AMP production by forskolin in pineals collected either early in the dark period or late in the dark phase. These data indicate that at least part of the action of extracellular calcium is indirect and that it affects steps in the induction of melatonin synthesis beyond the accumulation of cAMP.

Animals↗