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PubMedDate not supplied

Presence of immunoreactive neurophysins of a higher molecular weight in addition to the 10.000 form in the ovine pineal gland.

Using an aqueous extraction followed by ultrafiltration through Amicon Diaflo membranes, two ovine pineal fractions were obtained, which contain immunoreactive neurophysin. The presence of neurophysin was monitored by radioimmunoassay, employing an antiserum raised against pituitary bovine neurophysin and selected because it reacts with neurophysins of many other mammals. From 50 g of wet ovine pineal glands 552 micrograms of immunoreactive neurophysins were obtained. About 5% of these immunoreactive neurophysins are eluted from three different Sephadex columns with an elution volume corresponding to Mr above 10,000 between bovine serum albumin and pituitary neurophysin. The remaining 95% of ovine immunoreactive pineal neurophysin (Mr 10,000) shares immunological and physico-chemical properties with highly purified bovine pituitary neurophysin used as a reference. From the results of gel filtration and affinity chromatography on LVP-Sepharose it was concluded that ovine pineal gland may contain a neurophysin precursor molecule in addition to the neurophysin Mr 10,000.

Animals↗
PubMed1989

Serotonin and melatonin contents in the pineal glands from different stocks and strains of laboratory rats.

In the present study the pineal gland was examined in 2 outbred stocks and 6 inbred strains of rats some of which were pigmented to varying degrees, to see whether inbreeding affects the variability and whether differences exist between albino and pigmented rats. The animals were kept under 12 h light: 12 h darkness (12 L:12 D) and killed 7 h after the onset of light and darkness, respectively. The parameters examined were pineal protein content, serotonin and melatonin levels and hydroxyindole-O-methyltransferase (HIOMT) activity. All the parameters examined revealed interstrain differences, independently of whether the data were expressed per pineal or per mg protein. The variation coefficients for the various parameters were relatively high. They were mostly smaller when the data were expressed per pineal rather than per mg protein. No striking differences existed between the variation coefficients in inbred and outbred rats. When pineal size and the melatonin-related parameters expressed per pineal were used to assess the melatonin-synthesizing capacity of the pineal glands, it was found that the outbred Wistar and Sprague-Dawley rats and the inbred LEWIS-derived (LEW/Han) rats, all of which were albinos, had the most active pineals. Intermediate activity was noted in the hooded E3/Han and BDE/Han and the albino BDII/Han rats. The smallest and least active pineals were found in the totally pigmented BN/Han and DA/Han rats. The results taken together show that different stocks and strains exhibit significant differences in pineal size and melatonin-forming capacity. Albino rats appear to have larger and more active pineals than pigmented rats.

Acetylserotonin O-Methyltransferase↗
PubMedDate not supplied

Circadian variations of vasopressin level and vasopressin-converting aminopeptidase activity in the rat pineal gland.

Vasopressin levels and vasopressin-converting aminopeptidase activity were measured in the rat pineal gland during the 24 hr light-dark cycle. A rhythmic variation in peptide levels and peptidase activity occurred. At the onset of light at 6.00 hr, the peptidase displayed a significant, short-lasting (approximately 3 hr) increase of about 35% in activity, while a decrease of 28% in pineal vasopressin levels was observed. The changes in peptidase activity and peptide level were not triggered by light per se, since they persisted to occur at the same time point in animals which were not exposed to light, indicating the circadian nature of the rhythmicity. These changes were specific to the pineal gland, since other tissues, like hippocampus and pituitary gland, did not show these daily variations. The data suggest a relationship between vasopressin levels and vasopressin-converting aminopeptidase activity.

Aminopeptidases↗
PubMed1994

Immunohistochemical studies on the coexistence of catecholamine-synthesizing enzymes and neuropeptide Y in nerve fibers of the porcine pineal gland.

The distribution of nerve fibers immunoreactive (IR) to catecholamine-synthesizing enzymes, tyrosine hydroxylase (TH), dopamine-beta-hydroxylase (D beta H), and phenylethanolamine-N-methyltransferase (PNMT) as well as neuropeptide Y (NPY), in the pineal gland of adult sows was investigated by the use of double-labeling immunohistochemistry. The porcine pineal gland was found to be supplied by TH-IR nerve fibers, the vast majority of which were also D beta H-positive. TH- and TH/D beta H-IR nerves were located mostly in the capsula and connective septa of the pineal gland, and their fine varicose branches terminated in the adherent parenchyma. NPY-IR fibers formed a loose network overlapping that was observed in the case of TH- and D beta H-IR fibers. Most of NPY-IR fibers were also positive for TH. The density of the fiber networks depended on the region of the gland. No PNMT-IR structures were observed.

Animals↗
PubMed2000

Differential adrenergic regulation of the circadian expression of the clock genes Period1 and Period2 in the rat pineal gland.

Precise temporal regulation of transcription is pivotal to the role of the mammalian pineal gland as a transducer of circadian and seasonal information. The circadian clock genes Per1 and Per2 encode factors implicated in temporally gated transcriptional programmes in brain and pituitary. Here we show that the nocturnal circadian expression of Per1 and Per2 in the rat pineal gland parallels that of serotonin N-acetyltransferase (NAT) mRNA, which encodes the rate-limiting enzyme of melatonin biosynthesis. This rhythm is dependent upon an intact sympathetic innervation. Increases in rPer1 (r indicates rat) and rPer2, as well as rNAT, expression during subjective night were blocked completely by superior cervical ganglionectomy (SCGX). In SCGX rats, the beta-adrenergic receptor agonist isoproterenol rapidly induced the rPer1 mRNA with dynamics very similar to its effect on rNAT mRNA. In contrast, isoproterenol was without effect on expression of rPer2 mRNA. These findings demonstrate that circadian pineal expression of both rPer1 and rPer2 is controlled by sympathetic afferent innervation, but whereas beta-adrenergic signalling regulates rPer1 and rNAT, an alternative route mediates sympathetic regulation over rPer2 expression.

Animals↗
PubMed1998

Developmental pattern of cell type-specific calretinin immunoreactivity in the postnatal gerbil pineal gland.

The postnatal development of immunoreactivity for the neuronal calcium-binding protein calretinin in the pineal gland of the Mongolian gerbil was investigated using immunostaining of serial semithin sections. Calretinin-positive pineal cells could readily be visualized from the day of birth (P0) onwards and coexpressed the intermediate filament (IF) protein vimentin. During the first half of the first postnatal week, many of the calretinin-/vimentin-positive cells were also immunopositive for synaptophysin and neuron-specific enolase (NSE) and thus corresponded to pinealocytes. However, the expression of calretinin in pinealocytes was only transitory and declined towards the end of the first postnatal week. Thereafter, calretinin immunoreactivity became restricted to vimentin-positive interstitial glial cells. Therefore, in the gerbil pineal gland, calretinin obviously is not required in mature pinealocytes but instead serves as yet unknown functions in interstitial cells. The unusual calretinin expression pattern adds to the notion that pineal interstitial cells differ from glial cells of other brain regions. This conclusion is also underlined by our present detection of the neuronal marker protein PGP 9.5 in interstitial cells during postnatal development.

Animals↗
PubMed1981

Modes of protein and peptide uptake in the pineal gland of the Mongolian gerbil: an ultrastructural study.

The possible existence of either a blood-brain barrier or a CSF-brain barrier was examined in the pineal gland of the Mongolian gerbil using the ultrastructural tracers, horseradish peroxidase (HRP) and microperoxidase (MP). The mechanism of protein and peptide transport within the pineal gland and its possible relationship to pineal concretions was also considered. Gerbils were injected with either MP or HRP intravenously (IV), or they received intraventricular (IVT) injections of HRP. The IV injections resulted in both MP and HRP movement through the endothelial cells of the gland by vesicular transport and by diffusion through the endothelial cells of the gland by vesicular transport and by diffusion through the endothelial intercellular junctions. Following the IVT injections, the tracer was demonstrated in the subarachnoid space as well as in the glial cells associated with the periphery of the gland. In addition, after the IVT injections, rounded enlargements of the intercellular space that resembled canaliculi were filled with reaction product. In both cases (IVT and IV), the reaction product was localized to the perivascular space, to the glial cells and pinealocytes, and to the intercellular spaces. More importantly, there was specific localization of the tracers in the vacuolated pinealocytes and in the pineal concretions. The results of this study demonstrate several significant findings: 1) neither a blood-brain barrier nor a CSF-brain barrier exists in the pineal gland of the gerbil, 2) localization of the tracers in pineal concretions indicates a relationship between these structures and protein and peptide storage within the gland, and 3) the presence of the tracers within canaliculi after the IVT injections suggests a possible mode of secretion of pineal substances into the CSF.

Animals↗
PubMed1983

Diurnal variation in the serotonin content and turnover in the pineal gland of the Syrian hamster.

Serotonin and 5-hydroxyindoleacetic acid were measured in the pineal gland of Syrian hamsters by high performance liquid chromatography over a period of 24 h. A distinct 24 h rhythm was detected for both indoles. Turnover studies revealed a higher rate of serotonin synthesis during the day than during the night. We therefore suggest that the serotonin content in the pineal gland of the Syrian hamster is regulated by changes in its synthesis rate, rather than by changes of serotonin catabolism, via N-acetyltransferase activity.

Animals↗
PubMed1979

Development of the innervation in the chicken pineal gland (Gallus gallus).

The innervation of the pineal gland has been studied during the embryonic development and the first 10 days after hatching. On day 17 of embryonic development, the first nerve fibers are observed in the pineal capsule. They appear at the stalk level and rise to locate mostly on the anterior side of the capsule. Some nerve fibers leave these nerve bundles to penetrate the gland and they situate in the connective septa (18 days of development). From day 19 of development onwards, nerve fibers locate only in the parafollicular layer. Cells that may be identified as neurons are found in the pineal parenchyma.

Animals↗
PubMed1989

Immunohistochemical demonstration and radioimmunoassay of melatonin in the mink pineal gland.

An antiserum raised against N-amino-3-propyl melatonin bound to a protein carrier was used to visualize melatonin by immunohistochemistry and to measure melatonin concentration by radioimmunoassay in the pineal gland of intact mink females killed throughout the 24 h cycle and females killed after a bilateral ablation of the cervical superior ganglion. Melatonin immunoreactivity revealed by immunofluorescence or by the peroxidase-antiperoxidase complex was observed in the cytoplasm of presumed pinealocytes of all the females. Circadian changes in pineal melatonin content were not visualized by immunohistochemistry; furthermore, immunoreactivity was also present in the pineal gland of the ganglionectomized females. However, the melatonin content measured by radioimmunoassay was significantly higher in the pineal gland from intact females killed during the night compared with that of intact females killed during the day or of ganglionectomized females. The discrepancy between the results obtained using the two methods may arise because immunohistochemistry can detect very small amounts of melatonin.

Animals↗
PubMed1981

Electrophysiological study of evoked electrical activity in the pineal gland.

Experiments were carried out in rats, unanesthetized, paralyzed and artificially respirated. The electrical activity from the pineal gland was recorded with bipolar electrodes. Field potentials were evoked in the pineal after peripheral (photic and sciatic nerve) and central (septal area, habenular complex and optic tract) stimulations. In general these potentials were biphasic with the exception of that evoked by the sciatic nerve and optic tract, which exhibited a complex response and a triphasic field potential, respectively. Bilateral sympathectomy did not modify the pineal evoked responses, but when the pineal stalk was sectioned all the responses were immediately suppressed after the lesion. On the basis of the above experimental data one could conclude that the bulk of the inputs to the pineal gland come through its stalk. At the present, the physiological significance of these findings is not clear.

Animals↗
PubMed1992

[The mechanism of the pineal gland in inhibiting sexual receptivity of female rats: (II) Does it concern progesterone?].

The present study was designed to find the role of progesterone in modulating the effect of pineal gland upon reproductive behavior in the female rat. Monosodium L-glutamate (MSG) was used as a neurotoxin to induce hypogonadal function. In the first part of the experiment, rats were divided into four groups: (1) normal, (2) pinealectomized (Px), (3) MSG-treated (MSG), and (4) pinealectomized MSG-treated (Px-MSG), as immature rats became mature, the IEC (Incidence of Estrous Cycle) in each group was determined by vaginal smear for three weeks. Serum samples for estimating the levels of progesterone were collected at estrous. Then, ovariectomy was performed. Two weeks later, estradiol was primed for three days following progesterone administration and sexual receptivity as well as solicitation were observed. The results showed that, the serum progesterone levels and IEC of Px-MSG rats were significantly higher than that of the MSG group. In addition, Px and Px-MSG rats showed a higher sexual receptivity than normal and MSG rats, respectively. In the second part of the experiment, rats were divided into two groups: (1) normal and (2) Px. All rats were ovariectomized at 2.5 months of age and implanted with a 2-mm 20% estradiol benzoate-filled silastic capsule. Then, sexual receptivity to various doses of progesterone (0, 0.02, 0.1, 0.5 mg/rat) were observed. The results showed that receptivity of Px rats was significantly higher than that of the normal ones in a dose-dependent manner. It is concluded that pineal gland plays an important role in the regulation of ovary function as well as sexual receptivity in female rats. Also, progesterone may be one of the factors modulating the effect of pineal gland on sexual receptivity.

Animals↗
PubMed1998

Local renin-angiotensin system in the pineal gland.

Besides the classical endocrine renin-angiotensin system (RAS), a local RAS has been described also in the brain. We attempted to clarify the existence of a local RAS in the pineal gland. Through the use of a ribonuclease protection assay, it proved possible to detect the mRNA for angiotensinogen (AOGEN), for the angiotensin receptor type 1A (AT1a) and 1B (AT1b) and for the angiotensin-converting enzyme (ACE) in pineal glands from rats. Renin mRNA, however, could not be found by this method. By in situ hybridization and immunocytochemistry, AOGEN mRNA was co-localized with the astrocyte marker glial fibrillary acidic protein. AT1b mRNA expression exceeded the expression of AT1a mRNA and was co-localized with the pinealocyte-specific tryptophan hydroxylase. Thus, in the mammalian pineal gland there is a local formation of the components of the RAS. The presence of angiotensin II receptors further substantiates a role for angiotensins and the pineal RAS in the physiology of this gland.

Angiotensinogen↗
PubMed1987

Alpha 2-adrenergic regulation of arylalkylamine N-acetyltransferase in organ-cultured chick pineal gland: characterization with agonists and modulation of experimentally stimulated enzyme activity.

In the chicken pineal gland, norepinephrine, released at sympathetic nerve endings, plays a role in synchronizing the circadian rhythm of melatonin synthesis. This effect appears to be exerted via an adrenergic inhibition of arylalkylamine N-acetyltransferase, the melatonin rhythm-generating enzyme. The present study indicates that the nighttime peak of N-acetyltransferase activity developed by organ-cultured chick pineal glands is inhibited by adrenergic agonists with a potency order characterizing alpha 2-adrenergic receptors: UK 14,304 greater than clonidine greater than alpha-methylnorepinephrine = epinephrine greater than cirazoline greater than phenylephrine greater than isoproterenol. The mechanism of this alpha 2-adrenergic response was further analyzed in organ cultures, by studying the ability of clonidine to block the cyclic AMP-dependent and the depolarization-dependent stimulations of N-acetyltransferase activity. Clonidine prevented the rise in N-acetyltransferase activity evoked by the adenylate cyclase activators forskolin and cholera toxin or by the phosphodiesterase inhibitor Ro 20,1724. The stimulatory effect of dibutyryl cyclic AMP was also blocked by clonidine. Activation of pineal alpha 2-adrenergic receptors effectively prevented the stimulation of N-acetyltransferase by depolarizing concentrations of KCl. The possibility that the alpha 2-adrenergic effect might be exerted at a step distal to cyclic AMP production is discussed.

4-(3-Butoxy-4-methoxybenzyl)-2-imidazolidinone↗
PubMed1993

A comparative study of annual changes in the pineal gland morphology with reference to the influence of melatonin on testicular activity in tropical birds, Psittacula cyanocephala and Ploceus philippinus.

The aim of the present comparative investigation was to examine annual pineal cytological changes and the action of melatonin in relation to testicular activity in two wild tropical avian species. The findings revealed that in both blossomheaded parakeets (Psittacula cyanocephala) and Indian weaver birds (Ploceus philippinus) the pineal gland was inactive, with reduced karyometric values during the breeding season. A pinealoactive phase showing significantly increased pinealocyte nuclear diameter corresponded with the nonbreeding phase. Administration of melatonin (250 micrograms/100 g body wt) for 10 consecutive days caused significant involution of testes during the breeding phase, although it failed to alter reproductive activity during the nonbreeding phase. The results indicated that the pineal gland activity varied inversely with the seasonal testicular weight cycle and the antigonadal influence of melatonin appeared to be phase dependent, which corroborated the inverse temporal relationship of the pineal gland and male reproductive activity examined in the two tropical avian species.

Acclimatization↗
PubMed2000

Negative regulation of cytosolic phospholipase A(2) by melatonin in the rat pineal gland.

In this paper evidence that supports a new role for melatonin as a negative endogenous regulator of cytosolic phospholipase A(2) (cPLA(2)) is presented. When rat pineal glands were incubated in culture, time-dependent release of arachidonic acid (AA) was observed, which was significantly inhibited by a known 85-kDa cPLA(2) inhibitor, methyl arachidonyl fluorophosphonate. Co-incubation with melatonin inhibited the AA release in a concentration-dependent manner, and this decrease was accompanied by a reduction of cPLA(2) protein and mRNA expression. Melatonin-receptor agonists, 2-iodo-N-butanoyl-5-methoxytryptamine and 5-methoxycarbonylamino-N-acetyltryptamine, also decreased AA release and cPLA(2) protein and mRNA levels, while pre-incubation with the melatonin receptor antagonists luzindole and 2-phenylmelatonin abolished the melatonin effect. In vivo, as melatonin production reflected a typical diurnal variation, endogenous non-esterified AA and cPLA(2) mRNA levels in the rat pineal gland showed an off-phase diurnal pattern in relation to melatonin levels. Intravenous administration of isoproterenol, which has been shown to elevate melatonin production, also decreased the levels of non-esterified AA and cPLA(2) mRNA significantly. Direct administration of melatonin to rats by intravenous injection decreased the levels of non-esterified AA, cPLA(2) protein and mRNA in rat pineal glands. In conclusion, melatonin endogenously down-regulates cPLA(2) expression, presumably through melatonin-receptor-mediated processes.

Amino Acids↗
PubMed1993

Immunocytochemical and electron-microscopic characterization of macrophage/microglia cells and expression of class II major histocompatibility complex in the pineal gland of the rat.

Interstitial cells in the pineal gland of the rat were characterized immunocytochemically using the monoclonal antibodies MRC OX-42 and ED1 for macrophages/microglia, and MRC OX-6, which recognizes major histocompatibility complex (MHC) class II antigen. A polyclonal antibody against GFAP was used to identify astrocytes. Cells immunopositive for OX-42 and/or ED1 were distributed throughout the gland; they extended processes primarily along the perivascular spaces and occasionally within the parenchyma of the gland. Ultrastructurally, these OX-42-positive cells were characterized by a nucleus with sparse heterochromatin and cytoplasmic vacuoles/lysosomes. Cells expressing MHC class II antigen had a distribution and morphology similar to OX-42-immunopositive cells, suggesting that pineal macrophages/microglia play a role as antigen-presenting cells. GFAP-positive astrocytes were concentrated at the proximal end of the pineal where the pineal stalk enters the gland. The occurrence of antigen-presenting cells in this circumventricular neuroendocrine gland has important functional implications as these cells may be mediators of neuroimmunomodulatory mechanisms, and involved in certain disease states such as autoimmune pinealitis.

Animals↗