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The role of the pineal gland in magnetobiological effects.

The problem of pineal mognetosensitivity is considered in light of modem magnetobiology publications and the authors' own findings. It has been shown that electromagnetic exposure of different parameters affects pineal activity. Changes in the temporal organization of biological systems are suggested to be the main effect of electromagnetic exposure. Modern data evidence that there are magnetoreceptive structures aside from the pineal gland.

Animals↗

Expression of N-methyl-D-aspartate (NMDA) and alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) GluR2/3 receptors in the developing rat pineal gland.

The expression of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) type glutamate (GluR2/3) receptors and N-methyl-D-aspartate receptor subtype 1 (NMDAR1) was carried out by immunohistochemistry, double immunofluorescence and real-time RT-PCR analysis in the pineal glands of 1-day to 6-wk-old rats in the present study. GluR2/3 immunopositive cells were distributed throughout the pineal gland and showed branching processes in all age groups. The NMDAR1 immunoreactivity, however, was observed in fewer branched cells. A constitutive mRNA expression of NMDAR1, GluR2 and GluR3 was detected in the pineal glands of various ages and showed no significant difference between the age groups studied. Immunohistochemical and double immunofluorescence results showed that the GluR2/3 were mainly expressed and co-localized with OX-42-positive microglia/macrophages and the glial fibrillary acidic protein (GFAP)-positive astrocytes. Co-localization of NMDAR1 with OX-42- and GFAP-positive cells was much less. The expression of these receptors on the glial cells suggests that they may be involved in the development and growth of the pineal gland in the early postnatal period (1 day to 3 wk) and subsequently in the regulation of melatonin synthesis.

Animals↗

Parietal eye of the lizard: neuronal photoresponses and feedback from the pineal gland.

The parietal eye of the lizard responds to illumination by sending afferent impulses to the pineal gland during daylight, the photophase. The pineal gland has efferently conducting neurons which are especially sensitive to norepinephrine and whose feedback to the parietal eye enhances its photo responsiveness. During the scotophase, at night, the eye generates afferent impulses to the cessation of light and the pineal efferents are most sensitive to serotonin. Thus, the photo-and chemoresponses of this system of interacting neurons are nearly reversed during the two phases of the daily photoperiod of the lizard.

Action Potentials↗

Melatonin synthesis in the pineal gland of the Richardson's ground squirrel (Spermophilus richardsonii): influence of age and insulin-induced hypoglycemia.

The nocturnal rises in pineal N-acetyltransferase (NAT) activity and melatonin levels were compared in young (25-35 days old) and adult (at least 1 year old) Richardson's ground squirrels. When expressed as NAT activity per pineal gland, the nighttime rise in the activity of this enzyme was less in young than in the adult animals; conversely, the melatonin content of the pineal glands of young animals was higher at one point (4 a.m., 8 hours after darkness onset) when compared to that in adult squirrels. When data were expressed relative to total protein, the NAT and melatonin rhythms in the pineals of young and adult animals were very similar. The effect of insulin-induced hypoglycemia on both daytime and nighttime NAT and melatonin levels in the pineal gland of the Richardson's ground squirrel was also assessed. Low daytime levels of these constituents were not influenced by the administration of 10 units insulin, a treatment which caused a marked drop in circulating glucose levels. At night, when pineal NAT and melatonin levels were high insulin injection had a very modest stimulatory effect on NAT activity (one point was elevated above saline injected controls) while melatonin levels remained unchanged by the treatment. These findings in the ground squirrel in reference to insulin-induced hypoglycemia, and stressors in general, appear to differ from those in the rat where stress can have a substantial influence or both low daytime and high nighttime levels of pineal NAT and melatonin.

Aging↗

Pineal gland and polyamines.

The activity of ornithine decarboxylase was assayed in several organs (thymus, testes, prostate gland, liver, kidneys, adenohypophysis, anterior hypothalamus, and adrenals) taken from adult male rats killed at seven day interval up to six weeks after pinealectomy. The absence of the pineal gland particularly influences the ornithine decarboxylase activity in the thymus, in which the level of the enzyme is decreased irreversibly by the fourth week after the operation. In other organs the ornithine decarboxylase activity was often significantly different from that of corresponding shampinealectomized controls at various weeks after the surgical removal of the gland. Moreover, these differences between operated and sham-operated animals are sometimes positive, sometimes negative. Thus, the polyamine biosynthetic pathway in different organs appears to be regulated, directly or indirectly, by a new neuroendocrine centre, the pineal gland, and this pineal gland has thus been shown to be active in adult life in correlating endocrine-enzymatic functions.

Adrenal Glands↗

Presence of oxytocinergic neuronal-like cells in the bovine pineal gland: an immunocytochemical and in situ hybridization study.

In the last decade, there is more and more evidence showing the role of the central innervation of the pineal gland, but there are controversies around the intra or extrapineal origin of oxytocin found within the pineal tissue. In order to check the amount and the site of synthesis of oxytocin in the bovine pineal gland, we performed a morphological and chromatographic study. The anatomical distribution of the pineal oxytocin was explored by immunohistochemistry and in situ hybridization for the corresponding mRNA. The results confirm the presence of oxytocinergic fibres in the bovine pineal, some of them endowed with big varicosities. Immunohistochemistry also displayed neuronal-like cells in the pineal body. The in situ hybridization for the mRNA encoding pre-pro/oxytocin-NFZ I used a mixture of three oligonucleotide probes labelled with (35)S. This allowed identification of positive cells in the bovine pineal. The content in oxytocin was evaluated by radioimmunoassay during 5 months, from July to November, and the peptidic extract revealed an increase of pineal oxytocin immunoreactivity in September as compared with July or November. The significance of intrinsic oxytocin innervation of the bovine pineal gland, as well as the threefold increase of the oxytocin content in the pineal in September, remains to be elucidated.

Animals↗

Presence of melanin in the cat pineal gland.

Light- and electron-microscopic features of pigmented cells in the cat pineal gland are described. These cells were observed throughout postnatal life from the second postnatal day to the oldest cats studied (up to 13 years old). No apparent relationship was observed among the amount of pigment and the animal age or sex. Pigmented cells showed a preferential localization at the ventral surface of the pineal gland near its distal end. The pineal pigment was histochemically identified as melanin. Pineal pigment granules showed ultrastructural features similar to melanocyte melanin granules.

Animals↗

Forskolin, an activator of adenylate cyclase activity, promotes large increases in N-acetyl transferase activity and melatonin production in the Syrian hamster pineal gland only during the late dark period.

The exposure of organ cultured pineal glands of Syrian hamsters to forskolin, an adenylate cyclase activator, caused marked increases in serotonin N-acetyltransferase activity and melatonin content in a dose-related manner (1-100 microM) when glands were collected in the second half of the dark period. However, addition of forskolin to glands collected anytime during the light period or at the beginning of the dark period failed or only modestly stimulated either pineal N-acetyltransferase activity or melatonin levels. Similar results were obtained with isoproterenol. The results suggest that intrapinealocyte regulatory mechanisms may determine the nocturnal rise in the Syrian hamster pineal gland.

Acetyltransferases↗

Avian melatonin synthesis: photic and circadian regulation of serotonin N-acetyltransferase mRNA in the chicken pineal gland and retina.

The circadian rhythms in melatonin production in the chicken pineal gland and retina reflect changes in the activity of serotonin N-acetyltransferase (arylalkylamine N-acetyltransferase; AA-NAT; EC 2.3.1.87). Here we determined that the chicken AA-NAT mRNA is detectable in follicular pineal cells and retinal photoreceptors and that it exhibits a circadian rhythm, with peak levels at night. AA-NAT mRNA was not detected in other tissues. The AA-NAT mRNA rhythm in the pineal gland and retina persists in constant darkness (DD) and constant lighting (LL). The amplitude of the pineal mRNA rhythm is not decreased in LL. Light appears to influence the phase of the clock driving the rhythm in pineal AA-NAT mRNA in two ways: The peak is delayed by approximately 6 h in LL, and it is advanced by > 4 h by a 6-h light pulse late in subjective night in DD. Nocturnal AA-NAT mRNA levels do not change during a 20-min exposure to light, whereas this treatment dramatically decreases AA-NAT activity. These observations suggest that the rhythmic changes in chicken pineal AA-NAT activity reflect, at least in part, clock-generated changes in mRNA levels. In contrast, changes in mRNA content are not involved in the rapid light-induced decrease in AA-NAT activity.

Amino Acid Sequence↗

Plasma melatonin, pinealocyte morphology, and surface receptors/antigen expression on macrophages/microglia in the pineal gland following a high-altitude exposure.

The present study examined the effects of high-altitude exposure on the pineal gland, the main source of production of melatonin. It was surmised that hypoxia experienced at high altitude, caused by decreased oxygen tension in the ambient air, might lead to some structural alterations in the pineal gland and, hence, affect its melatonin production. Adult Wistar rats were exposed to an altitude of 8,000 m for 2 hr in an altitude chamber and then sacrificed at various time intervals after the exposure. Normal rats kept at ground level were used as controls. Blood samples were collected at various time intervals for measurement of plasma melatonin level, and the pineal glands from both groups were processed for electron microscopy and immunohistochemistry. The plasma melatonin level showed a steady increase following altitude exposure peaking at 7 days and returned to control levels thereafter. Between 1 and 4 days after altitude exposure, the mitochondrial number and lipid droplets in the pinealocytes appeared to be reduced compared with those in control rats. At 7 days, however, the mitochondrial numbers and lipid droplets were noticeably increased. At the same time interval, the expression of complement type 3 receptors and major histocompatibility class II antigens as detected with the antibodies OX-42 and OX-6, respectively, in macrophages/microglia was up-regulated compared with that in the control rats and those killed at earlier times. This was attributed to the increased serum melatonin after the altitude exposure. By 14 and 21 days, the ultrastructure of pinealocytes and immunoreactivity of macrophages/microglia were comparable with those in the control rats. We conclude from this study that an altitude exposure in rats leads to an increase in melatonin production, which returned to control levels with passage of time.

Altitude Sickness↗

Innervation of the rat pineal gland by PACAP-immunoreactive nerve fibers originating in the trigeminal ganglion: a degeneration study.

In order to establish that the pineal gland is innervated by pituitary adenylate cyclase-activating polypeptide (PACAP)-immunoreactive nerve fibers originating in the trigeminal ganglion, ophthalmic and maxillary nerves were transected by using a subtemporal fossa approach. The number of PACAP-immunoreactive nerve fibers in the pineal gland of rats with a total transection of the nerve was compared with that of rats without surgery. In the operated rat, PACAP-immunoreactive nerve fibers in the superficial pineal decreased remarkably, indicating that the trigeminal ganglion was the origin of these nerve fibers. This research provides evidence supporting the hypothesis that PACAP-immunoreactive nerves regulate the synthesis and/or secretion of melatonin in the pineal gland.

Animals↗

Calcium binding proteins immunohistochemistry and identification of neurons in the mammalian pineal gland of the African giant rat: Cricetomys gambianus.

The presence of true neurons in the rodent pineal gland is still a matter of controversy. In this work, by using immunohistochemistry with five antibodies against calcium-binding proteins (calbindin-D28k, calretinin, calmodulin, neurocalcin and S-100 beta) and Cricetomys gambianus, a rodent belonging to Muridae family living in Africa, we were able to illustrate the presence of neurons in the pineal gland. Anti-calbindin-D28k and anti-calretinin labelled neurons belonging to two neural ganglia. One ganglion was localized in the anterior part of the gland near the pineal stalk and the other one in the posterior portion of the organ. Immunoreactive neurons are medium in size (15-20 microns) and have long thick processes running towards the stalk. Calretinin and calbindin-D28k positive neurons stained with different intensities. Thin processes were detected by anti-calretinin whereas thick processes were preferentially calbindin-D28k positive. Neurocalcin labelled a few smaller neurons and many thin processes within the ganglion. Calmodulin could not be detected immunochemically. Within the ganglia many astrocytic processes were S-100 beta positive. The afferent and the efferent pathways of the pineal ganglia remain to be elucidated.

Animals↗

Distribution of NADPH-diaphorase activity in the pineal gland of the Turkey.

NADPH-diaphorase activity was histochemically demonstrated in the nerve fibers, neuronal-like cell bodies and in the endothelial cells of the vasculature in the pineal gland of the turkey. The nerve fibers were localized in the choroid plexus, connecting the pineal gland with the diencephalon as well as inside the pineal gland, where they formed basket-like structures around the pineal follicles. A group of neuronal-like cell bodies was observed in the proximal part of the gland. The positive staining was not observed in the pinealocytes of rudimentary-photoreceptor type and in the supporting cells.

Animals↗

Pineal gland involvement in retina-induced experimental allergic uveitis.

A form of experimental allergic uveitis (EAU) can be elicited in guinea pigs by injection of an extract of homologous retina homogenate (S preparation). These guinea pigs exhibit clinical and histological ocular disease as well as cellular and humoral immune responses to S preparation. Recent studies have shown an immunofluorescent crossreactivity of the sera of these animals (anti-S) with retinal photoreceptors and pinealocytes. The experiments presented in this paper demonstrate pineal gland involvement during the course of S preparation-induced EAU. There is lymphocytic infiltration of the pineal gland, with corresponding loss of antigen reactivity and some disturbance in cell structure. Pineal gland infiltrate can be seen before observable retinal involvement. Similarity of the reactive substances in retinal photoreceptors and pinealocytes is demonstrated by a reaction of identity in immunodiffusion studies and similar mobilities in immunoelectrophoretic studies of S preparation and pineal gland extract with anti-S serum. The relationship of the retinal and pineal substances to each other, as well as their role in disease development in the animal model and possibly in human uveitides, are discussed.

Animals↗

Pinopsin mRNA levels are significantly elevated in the pineal glands of chickens carrying a null mutation in guanylate cyclase-1.

The purpose of this study was to determine if the absence of guanylate cyclase-1 (RetGC1, GC1), a key visual phototransduction cascade enzyme that is expressed in both retinal photoreceptors and pinealocytes, disrupts light regulation of pinopsin mRNA levels in the chicken pineal gland. In this series of experiments, we compared levels of pinopsin and tryptophan 5-hydroxylase mRNA in the pineal glands of GUCY1*B (*B) and normal chickens housed under either cyclic light or constant dark conditions. The *B chicken carries a null mutation in the gene encoding guanylate cyclase-1 that results in blindness in these animals at hatching. The results of our experiments show (1) that the amount of pinopsin mRNA in *B pineal is significantly higher than the amount in normal pineal in both light and dark conditions, (2) that light induces an increase in pinopsin mRNA levels in *B pineal, (3) that the relative magnitude of the light-induced increase in pinopsin mRNA in *B pineal is not significantly different from that observed in normal pineal, and (4) that the changes in the regulation of pinopsin mRNA levels in *B pineal gland are not accompanied by changes in the circadian expression of tryptophan 5-hydroxylase mRNA. These results show that the absence of guanylate cyclase-1 expression in the *B pineal gland leads to a significant increase in basal levels of pinopsin mRNA in this gland but does not alter the magnitude of the increase in pinopsin mRNA levels that is observed as a result of light stimulation.

Animals↗

Adenyl cyclase activity in rat pineal gland: effects of chronic denervation and norepinephrine.

Adenyl cyclase activity in the pineal gland of rats was determined by measuring the rate of formation of radioactive cyclic 3',5'-adenosine monophosphate from (14)C-labeled adenosine triphosphate. Norepinephrine added in vitro to pineal homogenates enhanced this activity, while denervation of the pineal gland by superior cervical ganglionectomy did not significantly reduce it. The enzyme in these denervated glands was more responsive to the stimulatory effects of norepinephrine.

Adenine Nucleotides↗

Calcite microcrystals in the pineal gland of the human brain: first physical and chemical studies.

A new form of biomineralization has been studied in the pineal gland of the human brain. It consists of small crystals that are less than 20 microm in length and that are completely distinct from the often observed mulberry-type hydroxyapatite concretions. A special procedure was developed for isolation of the crystals from the organic matter in the pineal gland. Cubic, hexagonal, and cylindrical morphologies have been identified using scanning electron microscopy. The crystal edges were sharp whereas their surfaces were very rough. Energy dispersive spectroscopy showed that the crystals contained only the elements calcium, carbon, and oxygen. Selected area electron diffraction and near infrared Raman spectroscopy established that the crystals were calcite. With the exception of the otoconia structure of the inner ear, this is the only known nonpathological occurrence of calcite in the human body. The calcite microcrystals are probably responsible for the previously observed second harmonic generation in pineal tissue sections. The complex texture structure of the microcrystals may lead to crystallographic symmetry breaking and possible piezoelectricity, as is the case with otoconia. It is believed that the presence of two different crystalline compounds in the pineal gland is biologically significant, suggesting two entirely different mechanisms of formation and biological functions. Studies directed toward the elucidation of the formation and functions, and possible nonthermal interaction with external electromagnetic fields are currently in progress.

Adolescent↗

Light-induced suppression of nocturnal serotonin N-acetyltransferase activity in chick pineal gland and retina: a wavelength comparison.

Effects of white and monochromatic (blue-434 nm, green-548 nm, and red-614 nm) lights on the nighttime retinal and pineal NAT activity were examined in chicks. The potency of the tested lights to suppress NAT activity was similar for the retina and pineal gland, with a following rank order: white > green > blue > or = red. The studied tissues of chick were far less sensitive to pulses of monochromatic light than the rat pineal gland. The potency of light to decrease pineal NAT activity of rat was: white > green >> blue > red. In chicks, the suppression of the nocturnal NAT activity produced by a short 5-min pulse of monochromatic light was completely reversible in the pineal gland, and partially reversible in the retina. Our data suggest the existence of some differences between birds and mammals in terms of sensitivity and mechanisms involved in the light-induced suppression of melatonin biosynthesis.

Animals↗