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Diurnal variations of S-adenosyl-L-methionine and adenosine content in the rat pineal gland.

S-adenosylmethionine and adenosine levels in the rat pineal gland were determined by high-performance liquid chromatography after fractionation of the pineal extracts. The concentration of S-adenosylmethionine follows a circadian rhythm and is about three times higher during the day (2.5 nmol/gland) than the night (1.1 nmol/gland). The variations in the level of adenosine are apparently more complex. Over the 24 hours period there are two maxima at 03.00 (120 pmol/gland) and 15.00 hrs (100 pmol/gland) and one minimum at 09.00 hrs (50 pmol/gland). In addition, only an ultradian rhythm with a period of 12 hrs and an acrophase of 3 hrs can be evinced by computer analysis.

Adenosine↗

Immunocytochemical localization of serotonin in the pineal gland.

The localization of serotonin (5HT) in the rat pineal gland during daylight conditions was investigated by light and electron microscopy using a polyclonal antibody to 5HT and the immunocytochemical method of peroxidase-antiperoxidase (PAP). Thin varicose 5HT fibers were found distributed in the gland at light microscopy level. Ultrastructural observations revealed the presence of terminal endings containing 5HT and localized mainly in the perivascular spaces, labeled with dense diaminobenzidine (DAB) deposits.

Animals↗

Compensatory changes in contralateral sympathetic neurons of the superior cervical ganglion and in their terminals in the pineal gland following unilateral ganglionectomy.

The sympathetic noradrenergic neurons of the rat superior cervical ganglia (SCGs) provide the major source of innervation to the pineal gland. The present study sought to determine if this sympathetic innervation can undergo collateral sprouting following partial denervation of the pineal by unilateral removal of the SCG (ganglionectomy), and whether such growth of axon terminals is associated with biochemical changes in the contralateral SCG. In the pineal gland following partial denervation, residual noradrenergic terminals underwent compensatory changes indicative of collateral sprouting, as evidenced by: a rapid reduction in tyrosine hydroxylase (TH) activity and in [3H]norepinephrine (NE) uptake, to about 50% of control by 2 days, which was followed by a gradual but sustained increase to levels of approximately 80% of control by 10 days and a reduction in the intensity and density but not in the distribution of fibers containing NE-induced fluorescence by 2 days, which was followed by a sustained increase. In the contralateral SCG, choline acetyltransferase (CAT) activity, a marker of cholinergic preganglionic terminals, was transiently increased to about 115% of control by 4 days and returned to control levels by 14 days after unilateral ganglionectomy; later, TH activity in noradrenergic cell bodies was gradually increased to about 140% of control by 10 days where it remained for up to 52 days. Unilteral ganglionectomy combined with decentralization of the contralateral SCG by preganglionic nerve cut prevented the compensatory changes in noradrenergic nerve terminals within the pineal.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Molecular cloning and nucleotide sequence of cDNA encoding hydroxyindole O-methyltransferase of bovine pineal glands.

Messenger RNA for hydroxyindole O-methyltransferase (EC 2.1.1.4) was partially purified from poly(A)+ RNA isolated from bovine pineal glands by sucrose density gradient centrifugation. The enriched mRNA was used to prepare a cDNA library by use of expression vector lambda gt11. The library was screened with monoclonal antibodies to the enzyme, and three cDNA clones were isolated. These cloned cDNAs cross-hybridized with one another, and their fusion proteins reacted to the monoclonal antibodies with different binding properties. Hydroxyindole O-methyltransferase enzymatic activity was demonstrated in the bacteria lysate infected with lambda HIOMT-A16, the clone that contained the longest insert. An almost full-length cDNA clone was isolated from lambda gt10 cDNA library by use of the lambda HIOMT-A16 cDNA as a probe. The primary structure of hydroxyindole O-methyltransferase was determined by analyzing the nucleotide sequence of the cDNAs. It consisted of 1939 nucleotides including a 1050-nucleotide region coding for 350 amino acids. RNA transfer blot analysis indicated that mRNA encoding hydroxyindole O-methyltransferase was present only in the pineal gland and not in the brain, retina, and liver of cow.

Acetylserotonin O-Methyltransferase↗

Arginine vasotocin mRNA revealed by in situ hybridization in bovine pineal gland cells.

Arginine vasopressin (AVP) is the main antidiuretic hormone in mammals and arginine vasotocin (AVT) in submammalian vertebrates. The possibility that the genetic material encoding AVT is maintained in mammals is controversial. In this study, we investigated by radioactive in situ hybridization the possible presence of the mRNA encoding AVP and AVT, and using immunocytochemistry the presence of structures immunoreactive for AVP and AVT in the bovine pineal gland. In situ hybridization was performed by use of 35S-labelled oligoprobes. Immunocytochemistry was performed using specific polyclonal rabbit antibodies and the avidin-biotin-complex method. In situ hybridization revealed positive signals for both AVP mRNA and AVT mRNA in a few cells scattered throughout the pineal body. Immunocytochemistry revealed thin AVP-immunoreactive fibres in the pineal stalk and the pineal gland. It also revealed staining of several AVT-immunoreactive nerve fibres in both the pineal stalk and the gland. In addition, polyhedral, neuron-like cell bodies from which two to three processes emerged were also AVT-immunoreactive. Thus, our investigation shows the presence of AVP/AVT-immunoreactive cellular structures in the bovine pineal gland. Our data further show the presence of mRNAs encoding both AVT and AVP. We therefore suggest that AVT mRNA is translated into an AVT-like peptide in the bovine pineal.

Animals↗

Daily torpor alters multiple gene expression in the suprachiasmatic nucleus and pineal gland of the Djungarian hamster (Phodopus sungorus).

Circadian rhythms are still expressed in animals that display daily torpor, implying a temperature compensation of the pacemaker. Nevertheless, it remains unclear how the clock works in hypothermic states and whether torpor itself, as a temperature pulse, affects the circadian system. To reveal changes in the clockwork during torpor, we compared clock gene and neuropeptide expression by in situ hybridization in the suprachiasmatic nucleus (SCN) and pineal gland of normothermic and torpid Djungarian hamsters (Phodopus sungorus). Animals from light-dark (LD) 8ratio16 were sacrificed at 8 time points throughout 24 h. To investigate the effect of a previous torpor episode on the clock, we sacrificed a group of normothermic hamsters 1 day after torpor. In normothermic animals, Per1 peaked at zeitgeber time (ZT)4; whereas, Bmal1 reached maximal expression between ZT16 and ZT19. AVP mRNA in the SCN showed highest levels at ZT7. On the day of torpor, the levels of all mRNAs investigated, except for AVP mRNA, were increased during the torpor bout. Moreover, the Bmal1 rhythm was advanced. On the day after the hypothermia, Bmal1 and AVP rhythms showed severely depressed amplitude. Those distinct amplitude changes of Bmal1 and AVP on the day after a torpor episode expression suggests that torpor affects the circadian system, probably by altered translational processes that might lead to a modified protein feedback on gene expression. In the pineal gland, an important clock output, Aanat expression, peaked between ZT16 and ZT22 in normothermic animals. Aanat levels were significantly advanced on the day of hypothermia, an effect which was still visible 1 day afterward. In summary, this study showed that daily torpor affects the phase and amplitude of rhythmic clock gene and clock-controlled gene expression in the SCN. Furthermore, the rhythmic gene expression in a peripheral oscillator, the pineal gland, is also affected.

ARNTL Transcription Factors↗

The midline pineal "eye": MR and CT characteristics of the pineal gland with and without benign cyst formation.

The purpose of this study was to evaluate the magnetic resonance imaging characteristics of pineal cysts and pineal calcifications and to determine the incidence of benign pineal cysts. Two-hundred-fifty magnetic resonance examinations were retrospectively examined for the incidence of pineal cysts. In addition, 60 collected cases of pineal cysts were evaluated with regard to cross sectional diameter and magnetic resonance signal characteristics. Finally, the magnetic resonance signal characteristics of pineal tissue in 50 patients were compared to companion computed tomographic scans that were scrutinized for the presence or absence of calcification. The incidence of pineal cysts as revealed by magnetic resonance imaging in this study was 10.8%. The minimal and maximal measurements ranged from a low of 2 x 2 x 2 mm to a high of 10 x 12 x 10 mm. The magnetic resonance signal intensities of pineal cyst as compared to cerebrospinal fluid were iso- or hyperintense on all magnetic resonance sequences in the majority of cases. Calcifications of the pineal gland as revealed by computed tomography tended to be isointense to gray matter if the calcifications were small and hypointense to gray matter if large on all magnetic resonance acquisitions. A careful analysis of the magnetic resonance signal characteristics enables the recognition of moderate- to large-sized pineal calcifications and their differentiation from large pineal cysts. However, small cysts of the pineal gland can be difficult or impossible to distinguish on magnetic resonance imaging from calcifications without comparison with computed tomography.

Adolescent↗

Effects of superior cervical preganglionectomy on nerve vesicles and synaptic ribbons in the cat pineal gland.

Previous studies have shown that bilateral preganglionectomy of the superior cervical ganglia (SCGp) modifies the nerve vesicles and synaptic ribbons in the cat pineal gland. In the present study the nerve vesicles (NV) and synaptic ribbons (SR) were analyzed morphometrically after chronic preganglionectomy of the pineal sympathetic nerve fibers. It was shown that bilateral sympathectomy 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 chronic preganglionectomy 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.

Adrenergic Fibers↗

Neural control of the pineal gland.

The rhythm in melatonin production by the mammalian pineal gland is generated by the circadian timing system. The components of that system which mediate the function are: (1) visual projections through the retinohypothalamic tract to the circadian pacemaker, the suprachiasmatic nucleus (SCN) of the hypothalamus; (2) the SCN which generates a circadian signal transmitted by SCN projections to the parvocellular autonomic component of the paraventricular nucleus; (3) paraventricular nucleus projections to the upper thoracic intermediolateral cell column; (4) preganglionic sympathetic fibers to the superior cervical ganglion; (5) postganglionic sympathetic fibers from the superior cervical ganglion to the pineal.

Animals↗

Pineal gland lesions: a cytopathologic study of 20 specimens.

BACKGROUND: Pineal gland lesions are rare, with only a few cytologic descriptions occurring in the literature, according to the authors' knowledge. The current article describes the cytopathologic characteristics of 20 such lesions with discussion of differential diagnoses. METHODS: Cytologic material was obtained either by fine-needle aspiration biopsy (FNAB) under stearotactic radiologic guidance or by touch imprinting (TI) at the time of frozen sectioning. The 20 specimens include pineoblastoma (five specimens), pineocytoma (four specimens), astrocytoma (three specimens), germ cell tumor (three specimens), meningioma (one specimen), epidermoid cyst (three specimens), and pineal cyst (one specimen). Smears were stained with Diff-Quik and with Papanicolaou and hematoxylin and eosin stains. In selected specimens, immunoperoxidase (IPOX) stains were performed on cell block sections using synaptophysin, neuron-specific enolase, placental alkaline phosphatase, glial fibrillary acidic protein, leukocyte common antigen, cytokeratins, and human chorionic gonadotropin antibodies. RESULTS: Several cytomorphologic characteristics unique to each lesional category with occasional overlapping features were observed. The unique features included the following: small, hyperchromatic, round to oval cells with frequent rosetting (pineocytoma), with a few specimens in addition showing hypercellularity, crowding, mitoses, and necrosis (pineoblastoma); pleomorphic round cells in a fibrillary background (astrocytoma); large polygonal cells with prominent nucleoli and clear cytoplasm (germ cell tumor); spindled fibroblastic cells (meningioma); anucleate squames and mature squamous cells (epidermoid cyst); and small uniform polygonal cells (pineal cyst). When necessary, IPOX studies supported the morphologic diagnoses. CONCLUSIONS: FNAB and TI cytology were found to provide a rapid and reliable diagnosis of pineal lesions. This is particularly important when dealing with minute amounts of tissue material. Both techniques appeared to provide equally good cytomorphology on smears. IPOX studies played an important complementary role in difficult cases when performed on cell blocks.

Adolescent↗

Structural dissimilarities in different regions of the pineal gland of Pirbright white guinea-pigs.

In the present study, the proximal, intermediate and distal parts of dumbbell-shaped pineal glands of Pirbright White guinea-pigs were investigated with respect to structural dissimilarities. The most conspicuous difference was that while in the proximal part the pinealocytes were arranged in a random fashion, the intermediate and distal regions, except a small ventral area in the latter, exhibited a prominent follicular arrangement of the pinealocytes. Concretions were restricted to the subcapsular region of the intermediate part, whereas cells with hyperchromatic nuclei increased in number in an anterior-posterior direction. In males, pinealocyte nuclear size did not differ significantly in the regions investigated, whereas in females the karyometric results were much more heterogeneous. Exposure of male guinea-pigs to continuous illumination for 45 days resulted in (i) a 25% decrease of pineal volume, the intermediate area being relatively little affected, (ii) an increase in the number of follicles and (iii) a decrease in pinealocyte nuclear size, especially in a ventral zone of the proximal part of the pineal gland.

Animals↗

Some aspects of the pineal gland function.

Rhythmic production of melatonin by the mammalian pineal gland occurs in response to noradrenergic stimulation which produces a cascade of biochemical events within the pinealocyte. Melatonin is involved in a wide range of physiological processes, including seasonal reproduction and diurnal activity rhythms. It was recently discovered that melatonin is a very potent hydroxyl radical scavenger, reinforcing its relationship with the aging process and the immune system.

Animals↗

Daytime pineal gland activation in rats with colon tumors induced by 1,2-dimethylhydrazine(3).

OBJECTIVES: Intact rats and rats with 1,2-dimethylhydrazine induced tumors of large intestine were used in experiments. Previously, blood melatonin concentration in these tumor-bearing rats was shown to increase at night, but not in the daytime. METHODS: The extracellular microelectrode registration of rat daytime pineal gland activity was performed. RESULTS: The existence of different types of pinealocytes in the pineal gland was confirmed. Tumor-bearing rats, in comparison to intact, demonstrated higher spike frequency due to cells switching from regular to pattern (4-6 times gain) activity and appearance of "fast" cells (>5Hz frequency). CONSLUSIONS: The literature about pinealocytes points to the correlation between electrical and secretory processes in pinealocytes; thus we suppose the groups of interacting cells, detected in tumor-bearing rats, to reflect cascade cells activation while pineal gland secretion increases. The results indicate, that in the daytime pinealocytes are secreting substances (not melatonin) in dependence with hormonal background.

Journal Article↗

Maternal pineal gland participates in prepubertal rats' ovarian oocyte development.

BACKGROUND: Sexual maturation is a very complex phenomenom that it is mediated by the ontogeny of the hypothalamus-pituitary-gonadal axis during intrauterine life. The maternal pineal gland can affect fetal development because the main pineal hormone, melatonin, crosses the placental barrier. We found that melatonin treatment during gestation in the rat produced delayed sexual maturation of the female offspring. The present work was undertaken to study the maturational stage of oocytes of prepubertal female rats when their mothers were either pinealectomized (PIN-X) or treated with melatonin (MEL) during pregnancy. METHODS: Three groups of female Wistar rats were used: control, PIN-X, and those treated (250 micrograms/100 g body weight) with melatonin throughout pregnancy. Ovaries of 25-30- and 34-day-old female offspring were studied during the prepubertal phase. Morphometric studies of semithin sections (1 micron) of the ovaries were performed. Oocyte, nuclear, and nucleolar volumes were calculated by a computer-assisted program (M.I.P.) in an image analyzer Kontron. Regularity of the structures was determined by the frequency distributions of circular and regular form factors. RESULTS: Cytometric study of oocyte structure showed a frequency distribution of regular and circular form factors, with a high degree of regularity very close to unit. Cellular and nuclear volumes of follicular oocytes showed a transitory increase at 30 days of age in control rats. In the offspring of MEL-treated mother rats, a pattern of oocyte development showed significantly lower nuclear and nucleolar volumes at 30 days of age than at the other time points and significantly lower cellular volume at 34 days of age than at 25 days of age. In the offspring of PIN-X mother rats, no significant differences in oocyte cellular volumes were observed throughout prepubertal development, but we observed a significantly higher nuclear volume at 25 days of age and a significantly lower nucleolar volume at 30 days of age. CONCLUSIONS: These findings show that the maternal pineal gland participates in cellular and nuclear volumes of prepubertal oocyte development. Melatonin treatment during pregnancy resulted in a redirected postnatal oocyte development.

Animals↗

The ultrastructure of the human fetal pineal gland. II. Innervation and cell junctions.

The innervation of the pineal gland, the cell junctions in this organ and junctions between ependymal cells in the pineal recess were investigated in 27 human fetuses (crown-rump length 30-190 mm). Free nerve boutons containing clear and a few dense core vesicles were present in the pineal parenchyma and in the perivascular spaces. The boutons did not make "synaptic" contacts with the pinealocytes. No evidence for the presence of noradrenaline in the vesicles of nerve boutons was found. Gap junctions, intermediate-like junctions and desmosomes were frequently seen between the pinealocytes. Ruthenium red was used in three fetuses as an extracellular marker. The continuous endothelial cells surrounding the capillary lumen were connected by tight junctions. This indicates the presence of a blood-brain barrier. Tight junctions were present between the ependymal cells in the pineal recess. These junctions constitute an extracellular barrier between the pineal and the cerebrospinal fluid.

Axons↗

Expression and regulation of Icer mRNA in the Syrian hamster pineal gland.

Inducible-cAMP early repressor (ICER) is a potent inhibitor of CRE (cAMP-related element)-driven gene transcription. In the rat pineal gland, it has been proposed to be part of the mechanisms involved in the shutting down of the transcription of the gene coding for arylalkylamine N-acetyltransferase (AA-NAT, the melatonin rhythm-generating enzyme). In this study, we report that ICER is expressed in the pineal gland of the photoperiodic rodent Syrian hamster although with some difference compared to the rat. In the Syrian hamster pineal, Icer mRNA levels, low at daytime, displayed a 20-fold increase during the night. Nighttime administration of a beta-adrenergic antagonist, propranolol, significantly reduced Icer mRNA levels although daytime administration of a beta-adrenergic agonist, isoproterenol, was unable to raise the low amount of Icer mRNA. These observations indicate that Icer mRNA expression is induced by the clock-driven norepinephrine release and further suggest that this stimulation is restricted to nighttime, as already observed for Aa-nat gene transcription. Furthermore, we found that the daily profile of Icer mRNA displayed photoperiodic variation with a lengthening of the nocturnal peak in short versus long photoperiod. These data indicate that ICER may be involved in both daily and seasonal regulation of melatonin synthesis in the Syrian hamster.

Adrenergic beta-Antagonists↗

Alpha-adrenergic control of serotonin release from rat pineal glands.

This study was designed to determine if norepinephrine (NE) stimulated release of pineal serotonin (5-HT) is receptor mediated and to identify the receptor(s) most influential in the release process. Efflux of 3H-5-HT from rat pineals in vitro was significantly increased within 5 min after NE was added to perifusion buffer in concentrations ranging from 0.1 to 3 microM. 3H-5-HT was not simply displaced from pinealocytes by NE, since buffer containing 3 microM 5-HT had no effect on 3H-5-HT efflux. Furthermore, NE enhanced 3H-5-HT secretion by a stereospecific process, since d-NE was significantly less effective than l-NE. The alpha 1-adrenoreceptor agonists phenylephrine and cirazoline simulated the effects of NE, significantly increasing 3H-5-HT efflux in single and sequential stimulations. Furthermore, the alpha 1-adrenergic receptor antagonist prazosin reduced NE-stimulated release of 3H-5-HT. In contrast, the alpha 2- and beta-adrenoreceptor agonists naphazoline or clonidine and isoproterenol, respectively, were without effect. In addition, the alpha 2- and the beta-receptor antagonists rauwolscine and timolol, respectively, had no effect on NE-stimulated release of pineal 5-HT. In conclusion, the data show that NE stimulates 5-HT release within minutes from rat pineal glands in vitro. Unlike the pineal mechanism controlling melatonin synthesis, which has a longer latency and requires beta-adrenergic receptor activation, 5-HT release is regulated by activation of alpha 1-receptors. Thus, the pineal may be useful model for studying how separate intracellular processes are controlled by common neural stimuli.

Animals↗

Cytophysiological response of the pineal gland to long-term administration of norethisterone (a progestogen-only contraceptive) in rats.

Chronic administration of norethisterone for 75 days caused a marked inhibition of the pineal gland activity of cycling albino rats. In addition to a decrease in the nuclear diameter, possibly the cell size also decreased. The progestin, however, failed to induce mitosis in this gland. It is suggested that prolonged administration of norethisterone exerts untoward effects on the pineal gland function in adult female albino rats.

Animals↗