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Vesicular inhibitory amino acid transporter is expressed in gamma-aminobutyric acid (GABA)-containing astrocytes in rat pineal glands.

Gamma-aminobutyric acid (GABA) is an inhibitory amino acid and acts as an intercellular transmitter in the central nervous system and peripheral tissues. In pineal glands, GABA is supposed to be a paracrine-like modulator of secretion of melatonin, although its mode of action, especially the sites of GABA signal appearance, is unknown. Vesicular inhibitory amino acid transporter (VIAAT) is a potential marker for the GABAergic phenotype. Here we presented evidence that VIAAT is expressed in GFAP-expressing astrocytes and a subpopulation of OX42-expressing microglia, but not in pinealocytes in cultured cells of rat pineal glands. The VIAAT-expressing cells also exhibit GABA immunoreactivity. Essentially the same results were obtained for pineal glands. These results suggest that GABA is stored and secreted from astrocytes and a subpopulation of microglia in pineal glands.

Amino Acid Transport Systems↗

Tachykinins and tachykinin-receptors in the rat pineal gland.

High-pressure liquid chromatography of extracts of rat pineal glands, followed by radio immunological analysis with antibodies against tachykinins, demonstrated the presence of substance P, neurokinin A and neurokinin B in the superficial rat pineal gland. Immunohistochemistry on perfusion-fixed rat brain sections showed substance P and neurokinin A to be present in nerve fibers located both in the perivascular spaces as well as intraparenchymally between the pinealocytes. After extracting total RNA, followed by reverse transcription and polymerase chain reaction amplification with primers specific for NK1-, NK2- and NK3-receptors, agarose gel analysis of the reaction products showed the presence of mRNA encoding all three neurokinin receptors. Immunohistochemical analysis showed NK1 receptor to be located in the interstitial cells of the gland. This location was confirmed by use of in situ hybridization using radioactively labeled antisense oligonucleotide probes. Double immunohistochemical stainings showed that the NK1-immunoreactive cells were not a part of the macrophages or antigen-presenting cells of the gland. Our study suggests that tachykinins, after release from intrapineal nerve fibers, are involved in an up to now unknown function, different from that of melatonin synthesis.

Animals↗

The role of the pineal gland in the lipid metabolism.

The study contains the data gathered with time with regard to the involvment of the pineal gland in the lipid metabolism. Thus, it has been demonstrated that administration of the pineal extract lowers serum, hepatic, adrenal and testicular cholesterol of rat and that pinealectomy causes reverse effects. Administration of melatonin did not cause significant alterations. The same pineal extract reduced cholesterolemia, biliary cholesterol and serum phospholipids in rabbits whereas the absence of the pineal gland caused an increase in cholesterolemia, total lipids and FFA. Pineal treatment reduced the major lipid components of the blood and aorta in experimental atheromatosis induced in rabbits. The results presented in this paper demonstrate the inhibiting role played by the pineal gland in the lipid metabolism.

Adrenal Glands↗

Ultrastructural evidence of a secretory process in the rat pineal gland.

Pericapillar spaces of the rat pineal gland belong to the most active sites of this organ. Neighborhood of sympathetic nerve endings, capillaries and pinealocyte processes facilitates possibly the synthesis and the secretion of methoxyindoles. Lipid droplets migrate through the pinealocyte cellular processes towards the terminal enlargement or poles, where they are secreted into the pericapillar space, and the possibility that indoleamines are included in the lipid droplets has been discussed.

Animals↗

Cellular proliferation in the rat pineal gland during postnatal development.

To establish a possible correlation between the rate of cellular proliferation and already documented functional and morphological characteristics of the rat pineal gland during postnatal development, the bromodeoxyuridine labelling method was used to evaluate the fraction of cells at the S phase of the cell cycle in paraffin sections from 1-, 7-, 14- and 28-day-old rats. Numerical density, taken as an indirect measure of cell hypertrophy, was also evaluated. During the first week after birth the percentage of S phase-cells in the rat pineal gland sharply decreased from around 9% to 1.3%. A smaller but also significant decrease was found from the 7th to the 14th postnatal day where S phase cells were less than 0.5% of all pineal cells. A very low percentage was also seen in samples from 28-day-old rats. Numerical density, namely, the total number of cells per surface unit of pineal section, decreased from birth to the end of the first month. This decrease was also steeper from birth to the 7th postnatal day than at any other period of the study. These results support the idea that a strong expansion of the cellular population of the rat pineal gland precedes morphological and functional maturation and opens the way to further exploration of the relationship between functional and proliferative responses of the pineal gland.

Animals↗

Prolactin-releasing and release-inhibiting factor activities in the bovine, rat, and human pineal gland: in vitro and in vivo studies.

The effects of crude extracts of bovine, rat, and human pineal glands on prolactin (PRL) release were studied using an in vitro system. In addition, the effects of a known pineal constituent, arginine vasotocin (AVT), and crude bovine pineal extract (bPE) on PRL secretion were studied in vivo. Normal male rat hemipituitaries (HP), incubated with bPE (13 mg tissue/HP)released 200%, 150%, and 285% more PRL into the medium than did their corresponding untreated control halves incubated in either Medium 199 alone, hypothalamic extract, or cerebral cortical extract, respectively. HP incubated with either rat (6 mg of tissue/HP) or human (25 mg of tissue/HP) pineal extract released 110% and 75% more PRL, respectively, than did their corresponding untreated control halves. HP exposed to 10 mg tissue eq of either bovine pineal fraction A1 or bovine pineal fraction A3 released 88% and 63%, respectively, less PRL than did their corresponding untreated control halves incubated in Krebs-Ringer Bicarbonate (KRB) medium. Quantitites of melatonin, thyrotropin-releasing hormone (TRH), or estrogen, comparable to those found in the pineal, had no significant effect on PRL secretion in vitro. The iv injection of either bPE (90 mg tissue/rat) or AVT (10 mug/rat) into estrogen and progesterone-treated male rats resulted in a 40% and 138% increase, respectively, in plasma PRL titers, 10 min after injection, over pre-injection control levels. The per cent of increase in plasma PRL levels in these animals was significantly greater than that observed in control rats receiving either saline or cortical extract. The results suggest that crude extracts of pineal glands of three different species contain prolactin-releasing factor (PRF) activity which is probably not due to any endogenous melatonin, TRH, or estrogen that may be present. Conversely, two bovine pineal fractions, A1 and A3, appeared to exhibit prolactin-inhibiting factor (PIF) activity. We have concluded that the pineal gland may serve as an alternate or supplemental source of PRF and/or PIF.

Aged↗

Development of MEKA (phosducin), G beta, G gamma and S-antigen in the rat pineal gland and retina.

Pinealocytes and retinal photoreceptor cells contain an unusual cytoplasmic complex composed of the G beta gamma dimer of GTP-binding regulatory proteins (G-proteins) tightly bound to an acidic 33 kDa phosphoprotein termed MEKA or phosducin; MEKA is a substrate of cyclic AMP-dependent protein kinase. This study characterized the developmental appearance of these and two related proteins, G gamma and S-antigen, in pineal and retinal tissue. MEKA was absent in the pineal gland prior to birth, at a time when it was possible to detect G beta in pineal cytoplasm, indicating that the appearance of G beta in the cytoplasm precedes that of MEKA and does not appear to require the presence of MEKA. The absence of MEKA at this time indicates that the cyclic AMP stimulation of pineal serotonin N-acetyltransferase activity is not mediated by MEKA, which has been considered as a possible role of MEKA. After postnatal day 7, pineal MEKA and cytoplasmic G beta increased in a parallel manner, with peak values occurring at about postnatal day 21. Thereafter, both proteins in the pineal gland decreased in a parallel fashion to 10 and 35% of their peak values, respectively; in contrast, the cytoplasmic protein S-antigen and membrane associated G beta remained at maximal levels after this time. Whereas both MEKA and G beta decreased late in development in the pineal gland, these proteins either increased or remained constant in the retina. These tissue-specific patterns were found to differ from those of another cytosolic protein found exclusively in the pineal gland and retina, S-antigen, which remained constant after day 21 in the pineal gland but decreased in the retina late in life.

Aging↗

Sprouting of non-sympathetic myelinated and unmyelinated fibres in response to chronic sympathetic denervation in the pineal gland of the Chinese hamster, Cricetulus griseus.

We have examined the effects of chronic sympathetic denervation on non-sympathetic myelinated and unmyelinated fibres in the superficial pineal gland of the Chinese hamster (Cricetulus griseus), using LM, EM and immunohistochemistry. The results suggest that non-sympathetic, myelinated and unmyelinated fibres enter the superficial pineal gland at its distal portion by way of the nervi conarii, and that these fibres are immunoreactive for calcitonin gene-related peptide or substance P. Non-sympathetic, myelinated and unmyelinated fibres in the superficial pineal gland increased in number following chronic superior cervical ganglionectomy. The number of unmyelinated fibres in the nervi conarii also increased in ganglionectomized animals. Thus, the numerical increase of calcitonin gene-related peptide or substance P fibres found in the superficial pineal gland after long-term sympathectomy may be due to sprouting of these fibres. It is speculated that the growth of non-sympathetic, myelinated and unmyelinated fibres and myelination of the former fibres occurring after sympathectomy are caused by nerve growth factor-related mechanisms.

Animals↗

The pineal gland as an APUD organ: supporting evidence and implications.

It is only in recent years that the pineal gland has emerged from being thought of as non-functional and unimportant. The rise from obscurity has been the result of the interest of investigators of multidisplinary origins; such approaches, whilst clearly advancing understanding, also tend to leave knowledge fragmentary. In the last decade, a new neuroendocrine axis known as the APUD system has been delineated and scientifically accepted. The essence of the concept has been to link a physically widely dispersed and apparently unconnected series of glandular structures by a common embryological derivation and their secretion of closely similar functional products. The present proposal collates evidence for the inclusion of the pineal gland within the APUD system. In addition, because of the proven connections and response of the pineal gland to environmental changes, it is hypothesised that pineal products may provide the link between the environment and other members of the widely dispersed APUD system and thereby temper body homeostasis.

APUD Cells↗

Localization of kallikrein in rat pineal glands.

The presence of kallikrein mRNA has been reported in the pineal gland of rats. Using an antibody to rat tissue kallikrein, we immunohistochemically examined the localization of cell components producing tissue kallikrein in this gland. The kallikrein immunoreactive cells were scattered in the parenchyma of the pineal gland. Their cell bodies were polymorphic with cell processes and a large nucleus similar to that of the pinealocyte. Frequently immunoreactive materials were seen to be localized in the perivascular areas.

Animals↗

The human pineal gland and melatonin in aging and Alzheimer's disease.

The pineal gland is a central structure in the circadian system which produces melatonin under the control of the central clock, the suprachiasmatic nucleus (SCN). The SCN and the output of the pineal gland, i.e. melatonin, are synchronized to the 24-hr day by environmental light, received by the retina and transmitted to the SCN via the retinohypothalamic tract. Melatonin not only plays an important role in the regulation of circadian rhythms, but also acts as antioxidant and neuroprotector that may be of importance in aging and Alzheimer's disease (AD). Circadian disorders, such as sleep-wake cycle disturbances, are associated with aging, and even more pronounced in AD. Many studies have reported disrupted melatonin production and rhythms in aging and in AD that, as we showed, are taking place as early as in the very first preclinical AD stages (neuropathological Braak stage I-II). Degeneration of the retina-SCN-pineal axis may underlie these changes. Our recent studies indicate that a dysfunction of the sympathetic regulation of pineal melatonin synthesis by the SCN is responsible for melatonin changes during the early AD stages. Reactivation of the circadian system (retina-SCN-pineal pathway) by means of light therapy and melatonin supplementation, to restore the circadian rhythm and to relieve the clinical circadian disturbances, has shown promising positive results.

Aged↗

Two types of cells with central innervation in pineal gland of guinea pigs.

Cells within pineal glands isolated from young, male guinea pigs were impaled with intracellular microelectrodes and their responses to stimulate the nerve supply to the gland were studied. Two types of cells were identified. The response of cells of type I was a depolarization on which spikes were superimposed. Blockers of alpha-adrenoceptors abolished the spikes while beta-adrenoceptor blockers reduced the depolarization to 27%, leaving a small tetrodotoxin-sensitive depolarization. After bilateral removal of the superior cervical ganglia (SCG) the beta-mediated depolarization was not observed while the spikes and the smaller depolarization persisted. The response of cells of type II was an initial large, transient depolarization followed by a smaller depolarization. Both components were reversibly blocked by tetrodotoxin. The only agents found to have any effect on these cells were oxytocin, vasopressin, and vasotocin. These peptides caused depolarization similar in amplitude to the larger response to nerve stimulation, although more prolonged. The large depolarization was not observed following ganglionectomy, but the smaller one persisted. It is concluded that cells of type I and II both receive inputs from nerves whose cell bodies lie in the SCG. Cells of both types are also innervated through another pathway.

Action Potentials↗

[Symptomatic glial cysts of the pineal gland: report of two cases and review of the literature].

Referring to two individual cases, the authors review clinical, radiological and histological features of benign glial cysts of the pineal gland. Both patients were young females with aggravating headaches and with convulsions in one case. Symptoms were referable to a space-occupying cystic mass of the pineal gland. On histology, both lesions proved to be non neoplastic cysts without an epithelial lining. Their histogenesis and low growth potential were reinforced by immunohistochemical analysis of pineal antigens and proliferation markers. Glial cysts of the pineal gland are not infrequent, but symptomatic occurrences are exceptional. Most glial cysts are of dysontogenic or degenerative origin. Sometimes, however, the role of hormonal influences or paraneoplastic factors must be considered. Symptoms caused by glial cysts of the pineal gland are non-specific and radiologic imaging technics may contribute little to etiologic diagnosis. Pineal cysts are curable by surgical resection or stereotactic decompression. Whatever the diagnostic approach, emphasis must be laid on the histologic examination in order to avoid unnecessarily aggressive treatment.

Adolescent↗

Development of the circadian melatonin rhythm and the effect of PACAP on melatonin release in the embryonic chicken pineal gland. An in vitro study.

Pituitary adenylate cyclase activating polypeptide (PACAP) has been shown to participate in modulation of circadian rhythm and to stimulate melatonin (MT) secretion in both the rat and chicken pineal glands. In contrast to mammals, the main regulator of circadian rhythm in birds is the pineal gland, which begins its rhythmic MT production already during embryonic life. In the present study, we investigated the development of MT secretion in explanted embryonic chicken pineals and their responsiveness to PACAP in a perifusion system. Our results show that: (1) the circadian clock and/or the intracellular signal transduction system connecting the clock to MT synthesizing apparatus develop between the embryonic days 16-18 (E16-18), even in vitro. (2) Exposure of the embryonic chicken pineal gland to PACAP induces transitory increase in MT secretion but does not induce visible phase shift in the circadian rhythm. (3) Cyclic AMP (cAMP) efflux also responds to PACAP at or before day E13 in embryonic chicken pineal gland in vitro.

Animals↗

Some properties of pineal gland hydroxyindole-O-methyltransferase from black rhinoceros (Diceros bicornis).

Pineal glands were obtained from two young female black rhinoceri that had died as a result of postcapture trauma during a translocation exercise. Hydroxyindole-O-methyltransferase (HIOMT) from these pineal glands showed a peak activity at pH 8.2, although high activity extended over a fairly wide pH range (7.8-8.4). N-acetylserotonin was the best hydroxyindolic substrate for the enzyme, although other hydroxyindoles were methylated, the relative affinities being similar to values previously reported for bovine HIOMT. Kinetic analyses revealed that black rhinoceros HIOMT was subject to substrate inhibition by both substrates at high concentration; this observation is unlikely to have physiological significance. The catalytic mechanism was found to be ordered Bi-Bi, in which S-adenosylmethionine is the obligatory first substrate to bind to the enzyme, such binding allowing for binding of the hydroxyindolic substrate followed by catalysis, products again leaving the catalytic site in a sequential fashion.

Acetylserotonin O-Methyltransferase↗

Tumor-inhibiting activity in the rat pineal gland displays a circannual rhythm.

The aim of this study was to investigate whether the tumor-inhibiting activity present in the rat pineal gland undergoes seasonal fluctuations as do other pineal substances. Crude ethanol extracts of rat pineal glands were tested for tumor-inhibiting activity in an in vitro microbioassay using human erythroleukemia cells which could not be inhibited by melatonin. Highest activity was detected in summer and least inhibition and even stimulation were observed in winter. There were no differences in activity between animals of different age, sex, or strain. Therefore, season seems to be the factor that exerts the most important influence on the content of tumor-inhibiting activity in the rat pineal gland. Correlation with seasonality in the occurrence of cancer is discussed.

Animals↗

Characterization of gonadotropin-releasing hormone (GnRH)-immunoreactive protein in the rat pineal gland.

The aim of the present study was to characterize GnRH-like substance(s) in the rat pineal gland using a monoclonal antibody, LRH13, as a probe. The epitope of LRH13 is between 2nd and 5th amino acid residues of the mammalian GnRH, and its immunological characters were previously defined by us. LRH13 could show strong immunological signal on the rat pineal gland. Immunoblot after SDS-PAGE of the pineal gland preparations showed a LRH13 immunoreactive band with apparent mol wt 52 kilo-Dalton (kD), which is much bigger than that of hypothalamic GnRH precursor (10 kD). The 52 kD protein, however, was detected from insoluble fraction of the pineal homogenate and liberated from the fraction by Triton X-100 (2%) treatment. On the other hand, NaCl (140 mM and 500 mM) or EDTA (10 mM) treatment failed to liberate. Two-dimensional gel electrophoresis showed that the 52 kD protein is a mixture of two proteins with different isoelectric points (pI approximately 6.8 and 7.0). Both proteins showed identical patterns of peptide mapping by V8 protease digestion, and they might be originated from the same peptide. These results suggest that the rat pineal GnRH-immunoreactive substance has a unique property as a membrane associate protein.

Animals↗