Showing papers on "Pinealocyte published in 1995"
TL;DR: The rhythm in melatonin production by the mammalian pineal gland is generated by the circadian timing system, and the components of that system are described.
374 citations
TL;DR: To identify proteins that regulate light–dark variations in pineal function, a subtractive hybridization technique based on the polymerase chain reaction (PCR) was used to isolate rat pineal gland messages that are differentially expressed by day and night.
Abstract: Formation of the pineal gland hormone melatonin increases markedly at night in response to light-dark environmental alterations. Melatonin is synthesized from serotonin by an initial N-acetylation followed by methylation of the 5-hydroxy moiety by hydroxyindole-O-methyltransferase. Serotonin N-acetyltransferase (NAT; EC2.3.1.87), which catalyses the first reaction, is the rate-limiting enzyme in this process, and its activity increases dramatically with the onset of darkness. Because melatonin may play important biological roles in reproduction, ageing and sleep, understanding the molecular factors that regulate NAT is of particular importance. To identify proteins that regulate light-dark variations in pineal function, we used a subtractive hybridization technique based on the polymerase chain reaction (PCR) to isolate rat pineal gland messages that are differentially expressed by day and night. Here we report the molecular cloning of NAT and dramatic diurnal variations in its transcription. Independently, Klein and associates have cloned NAT from sheep pineal glands.
263 citations
Journal Article•
TL;DR: CAMP-dependent CREB phosphorylation could play a role in the adrenergic regulation of gene expression in pinealocytes, and in vivo studies show that CREBosphorylation can be induced by treatment with isoproterenol, demonstrating that phosphorylations of pineal CREB occurs in intact animals.
Abstract: Norepinephrine (NE) regulates melatonin production and many other aspects of pineal function through actions involving cAMP. In the present study the effects of NE on the phosphorylation of the cAMP response element-binding protein (CREB) were studied to determine whether CREB phosphorylation might be involved in cAMP signal transduction in this tissue. CREB was detected using gel mobility-shift analysis with the radiolabeled Ca2+/cAMP response element of the c-fos promoter. CREB phosphorylation was estimated in the gel mobility-shift assay using an antiserum specific for phosphorylated CREB. This antiserum generates a supershifted CREB signal with protein extracts obtained from glands treated with NE (EC50 approximately equal to 10 nM) in organ culture, demonstrating that NE stimulates CREB phosphorylation. CREB phosphorylation peaks 30-45 min after NE treatment is initiated and then gradually returns to base-line values. Pharmacological studies show that NE-stimulated CREB phosphorylation is mediated primarily through beta 1-adrenergic receptor-stimulated increases in cAMP. Activation of alpha 1-adrenergic receptors, which is known to elevate the intracellular free Ca2+ concentration, does not cause CREB phosphorylation. However, it is possible to produce CREB phosphorylation with certain pharmacological agents that elevate the intracellular free Ca2+ concentration. In vivo studies show that CREB phosphorylation can be induced by treatment with isoproterenol (1 mg/kg), demonstrating that phosphorylation of pineal CREB occurs in intact animals. These studies indicate that cAMP-dependent CREB phosphorylation could play a role in the adrenergic regulation of gene expression in pinealocytes.
155 citations
TL;DR: The biological clock in the suprachiasmatic nucleus of the hypothalamus plays a well-defined role in regulating melatonin production by the pineal, and emerging evidence indicates that melatonin itself can feed back upon the SCN and thereby influence circadian functions.
118 citations
TL;DR: Evidence is presented suggesting that activation of melatonin receptors in the SCN regulates circadian rhythms of behavior in the mouse, suggesting that melatonin may be important in synchronizing circadian rhythms to changes in the natural photoperiod.
90 citations
TL;DR: The data indicated IMEL binding in retinorecipient structures of the circadian, tectofugal, thalamofugal and accessory optic visual pathways in all avian species, but that this capacity has been lost in mammals.
Abstract: The roles that the pineal gland and its hormone melatonin play in the regulation of circadian rhythmicity and photoperiodism vary among vertebrate species. Recently, putative sites of melatonin action have been elucidated in several avian and mammalian species by application of in vitro binding of a radioiodinated melatonin agonist, 2[125I]iodomelatonin (IMEL) and autoradioradiography. These studies in mammals, birds and reptiles have indicated profound differences in the distribution of IMEL binding between these diverse groups, suggesting that these large differences in binding may reflect differences in melatonin function. The present study was performed to determine systematically whether the variance in IMEL binding among avian species corresponds to changes in circadian organization and/or phylogenetic relationships. The distribution of specific IMEL binding was determined in the brains from birds belonging to 14 different species in 5 Orders (Psittaciformes, Passeriformes, Columbiformes, Galliformes and Anseriformes) using in vitro binding, autoradiography and computer-assisted image analysis. The distribution was compared to a similar study in 3 species of turtles as an outgroup. The data indicated IMEL binding in retinorecipient structures of the circadian, tectofugal, thalamofugal and accessory optic visual pathways in all avian species. Relay nuclei and integrative structures of the tectofugal, thalamofugal, accessory optic, and limbic systems, however, bound the hormone to varying degrees. In turtles, binding was observed in retinorecipient structures of the thalamofugal visual pathway and in retinorecipient and integrative areas of the tectofugal visual pathway. No binding was observed in the pineal gland, tuberal hypothalamus or adenohypophysis in any avian or testudine species. This distribution is drastically different from that observed in mammals, where binding predominates in the pars tuberalis of the adenohypophysis and in the suprachiasmatic nucleus, suggesting that the circadian system may influence a wide array of sensory and integrative functions in birds and reptiles through the circadian secretion of melatonin, but that this capacity has been lost in mammals.
76 citations
TL;DR: D-Asp was found to be distributed in the cytosol of pinealocytes, but its biological role remains unclear, suggesting that biosynthesis of D- aspartic acid in the pineal gland occurs at night.
Abstract: We found a high concentration (1030 pmol per pineal gland) of D-aspartic acid (D-Asp) in the pineal gland of 6-week-old Sprague-Dawley (SD) rats. The content of D-Asp decreased with age, being 210 and 33 pmol per pineal gland in 28- and 45-week-old rats respectively. The proportion of D-Asp [(D-Asp/total Asp) x 100] also decreased with age, declining from 66% to 10% between 6 and 45 weeks after birth. The proportion of D-Asp did not differ between the sexes. The concentration of D-Asp was higher at night (at 2.00 a.m. 2830 +/- 485 pmol per pineal gland) than during the day (at 10.00 a.m. 1030 +/- 200 and at 3:00 p.m. 682 +/- 194 pmol per pineal gland), suggesting that biosynthesis of D-Asp in the pineal gland occurs at night. D-Asp was found to be distributed in the cytosol of pinealocytes, but its biological role remains unclear.
71 citations
TL;DR: The findings support the notion that phosphorylation of CREB is a rather rapid and uniform response of pinealocytes to noradrenergic stimulation and thus is an important link between adrenoreceptor activation and subsequent gene expression in the rat pineal organ.
Abstract: In the present study we investigated whether norepinephrine, which stimulates melatonin biosynthesis in the mammalian pineal organ, causes phosphorylation of the cyclic AMP responsive element binding protein (CREB) in rat pinealocytes Cells isolated from the pineal organ of adult male rats and cultured on coated coverslips were treated with norepinephrine, beta- or alpha 1-adrenergic agonists for 12, 5, 10, 20, 30, 60 or 300 min and then immunocytochemically analyzed with an antibody against phosphorylated CREB (p-CREB) Treatment with norepinephrine or beta-adrenergic agonists resulted in a similar, time-dependent induction of p-CREB immunoreactivity, exclusively found in cell nuclei The alpha 1-adrenergic agonist phenylephrine did not induce p-CREB immunoreactivity at low doses (01 microM) or when high doses (10 microM) were applied in combination with a beta-antagonist (propranolol, 01 microM) This indicates that induction of CREB phosphorylation is elicited by beta-adrenergic receptor stimulation The response was first seen after 10 min and reached a maximum after 30 to 60 min when more than 90% of the cells displayed p-CREB immunoreactivity The intensity of the p-CREB immunoreactivity showed marked cell-to-cell variation, but nearly all immunoreactive cells were identified as pinealocytes by double-labeling with an antibody against the S-antigen, a pinealocyte-specific marker The results show that norepinephrine stimulation induces p-CREB immunoreactivity by acting upon beta-adrenergic receptors in virtually all rat pinealocytes The findings support the notion that phosphorylation of CREB is a rather rapid and uniform response of pinealocytes to noradrenergic stimulation and thus is an important link between adrenoreceptor activation and subsequent gene expression in the rat pineal organ
70 citations
TL;DR: The results underline the pivotal role of norepinephrine for the regulation of pineal signal transduction, but they also support the notion that other neurotransmitters and neuropeptides are involved in the modulation of Pineal calcium signalling.
69 citations
TL;DR: CGMP modulates L-type Ca2+ channel currents in rat pinealocytes, causing inhibition of this current; the action of cGMP appears to be independent of cAMP elevation; and phosphorylation by cG MP- dependent protein kinase may be involved.
Abstract: In this study, the effect of cGMP on the dihydropyridine-sensitive (L- type) Ca2+ current was investigated using the whole cell version of the patch-clamp technique in rat pinealocytes Dibutyryl-cGMP (1 x 10(-4) M) induced a pronounced inhibition of the L-type Ca2+ channel current The dibutyryl-cGMP effect was concentration dependent Elevation of cGMP by nitroprusside had a similar inhibitory action on the L-type Ca2+ channel current Norepinephrine, which increased cGMP in rat pinealocytes, also inhibited this current The action of cGMP was independent of cAMP elevation since the cAMP antagonist, Rp-cAMPs, had no effect on the inhibitory action of dibutyryl-cGMP The involvement of cyclic GMP-dependent protein kinase was suggested by the blocking action of two protein kinase inhibitors, (1-(5-isoquinolinesulfonyl)-2- methylpiperazine (H7) and N-(2-guanidinoethyl)-5- isoquinolinesulfonamide (HA1004), on the dibutyryl-cGMP effect on the L- type Ca2+ channel current Taken together, these results suggest that (1) cGMP modulates L-type Ca2+ channel currents in rat pinealocytes, causing inhibition of this current; (2) the action of cGMP appears to be independent of cAMP elevation; and (3) phosphorylation by cGMP- dependent protein kinase may be involved
60 citations
Journal Article•
TL;DR: Observations indicate that fundamentally different mechanisms generate the responses to NE and VIP, with the dominant effect of VIP causing transient elevation of [Ca2+]i appearing to be through cGMP gating aI-cis-diltiazem-sensitive rod-type cyclic nucleotide-gated cation channel.
Abstract: Vasoactive intestinal peptide (VIP) receptor density is high in the pineal gland, which receives VIP innervation and responds to VIP with a relatively small increase in cAMP and cGMP levels. In the present study, we show that VIP (5-200 nM) treatment increased the intracellular calcium concentration ([Ca2+]i) in 64% of isolated individual pinealocytes; in comparison, norepinephrine (NE) elevated [Ca2+]i in 93% of the cells and produced more robust responses. Analysis of the role of second messengers indicated that [Ca2+]i was strongly elevated by cGMP analogs, but not by cAMP analogs. The nitric oxide-releasing agent S-nitro-N-acetylpenicillamine and 2,2-diethyl-1-nitroxyhydraxine also elevated [Ca2+]i. Investigation of the mechanisms revealed that responses to VIP or 8-bromo-cGMP involved Ca2+ influx, as did the plateau component of the response to NE; the large rapid component of the response to NE, however, appeared to reflect release from intracellular stores. Pharmacological studies indicated that the VIP-induced Ca2+ influx was mediated by a retinal rod-type cyclic nucleotidegated cation channel, expression of which was confirmed by reverse transcription-polymerase chain reaction analysis. These observations indicate that fundamentally different mechanisms generate the responses to NE and VIP. The dominant effect of VIP causing transient elevation of [Ca2+]i appears to be through cGMP gating aI-cis-diltiazem-sensitive rod-type cyclic nucleotide-gated cation channel. In contrast, the dominant effect of NE on [Ca2+]i is due to enhanced Ca2+ release from intracellular stores; the plateau component is due to influx through aI-cis-diltiazem-insensitive channel.
TL;DR: Results indicated that the MVs are organelles for storage of l‐glutamate in pinealocytes and suggested a possibility that pinealocyte transmit glutamate signals by MVs‐mediated exocytosis.
TL;DR: Mechanisms regulating melatonin synthesis in the avian pineal gland are established during embryonic life, and no rhythm of melatonin release was detected under constant dark (DD) conditions in a cell culture from 14-day-old chick embryos.
TL;DR: It is shown that PINX rats free-run with a longer period in four different light intensities than do SHAM rats, which supports the first hypothesis and further support a growing body of literature indicating that the pineal gland and its hormone melatonin play a role in mammalian circadian organization.
Abstract: Although removal of the pineal gland has been shown to have very little effect on the mammalian circadian system in constant darkness (DD), several recent reports have suggested that the mammalian pineal gland may be more important for circadian organization in nocturnal rodents than was previously believed. Removal of the pineal gland (PINX) facilitates the disruptive effects of constant bright light on wheel-running rhythmicity. This suggests at least two possibilities for the role of the pineal gland in the mammalian circadian system. First, pinealectomized rats may perceive ambient light intensity to be brighter than do sham-operated (SHAM) rats. Second, the pineal gland, probably via its secretion of melatonin, may also be involved in coupling components of the circadian system. Coupling, as we see it, may occur at several levels of organization: (1) between retinohypothalamic afferents and suprachiasmatic nuclei (SCN) oscillatory neurons, (2) among multiple SCN oscillators, (3) between the SCN and their multiple outputs, and/or (4) among the multiple circadian outputs themselves. In this study we show that PINX rats free-run with a longer period in four different light intensities than do SHAM rats. Moreover, the rate of increase of tau is greater among PINX rats than among SHAM rats. This supports the first hypothesis. We also show that in PINX rats the circadian rhythms of wheel running, general activity, body temperature, and heart rate are all more disrupted in constant bright light than are those of SHAM rats, and each rhythmic output is disrupted in parallel. This supports the second hypothesis. Melatonin is probably not involved in coupling presynaptic elements of SCN afferents in the retinohypothalamic tract to pacemaking cells within the SCN, since enucleation has no effect on SCN 2-[125I]iodomelatonin (IMEL) binding. Together the data do not discount either of the two hypotheses but do restrict the possible levels at which the pineal gland is involved in coupling. These data also further support a growing body of literature indicating that the pineal gland and its hormone melatonin play a role in mammalian circadian organization.
TL;DR: The results suggest that chick pineal cells from embryonic day 16 onwards are photosensitive but that the endogenous component of the melatonin rhythm is not completely developed at that age, and that the stimulatory and inhibitory pathways controlling melatonin synthesis in the mature pineal gland are effective.
Abstract: The chick pineal gland exhibits circadian rhythms in melatonin synthesis under in vivo and in vitro conditions. A daily rhythm of melatonin production was first detectable in pineal glands isolated from chick embryos at embryonic day 16 and incubated under a LD cycle. All pineal glands isolated from 17-day-old and older embryos were rhythmic while no gland isolated at embryonic day 14 and 15 exhibited a daily rhythm in melatonin synthesis. Melatonin production in static cultures of embryonic pineal cells was rhythmic over 48 h if the cells were kept under a LD cycle. When embryonic pineal cells were incubated in constant darkness the rhythm in melatonin production was damped within 48 h. These results suggest that chick pineal cells from embryonic day 16 onwards are photosensitive but that the endogenous component of the melatonin rhythm is not completely developed at that age. A soluble analogue of cAMP stimulated and norepinephrine inhibited melatonin synthesis in cultured embryonic pineal cells. These findings indicate that the stimulatory and inhibitory pathways controlling melatonin synthesis in the mature pineal gland are effective in pineal cells isolated from chick embryos at least 2 days before hatching.
TL;DR: Data support the view that NOS is a component of the cGMP‐generating system in mammalian pinealocytes and may have other important actions in the pineal, namely the inhibition of adrenergic‐stimulated melatonin synthesis.
Abstract: The presence of soluble guanylate cyclase in the pineal and its regulation by adrenergic pathways has been well documented. Recent evidence points to adrenergically stimulated nitric oxide generation as a mechanism for coupling this pathway. To what extent nitric oxide (NO) signalling can influence adrenergically stimulated melatonin synthesis has not been investigated. Cyclic guanosine 3',5'-monophospate (cGMP) signal transduction in the bovine pineal has also received little attention. We describe in the present report: 1) a dose-dependent elevation of cGMP in response to the nitrovasodilators, sodium nitroprusside (SNP) and 3-morpholino-sydnonimine (SIN-1), 2) a dose-dependent inhibition of melatonin synthesis by SNP and SIN-1, but not by 8-Br-cGMP in both bovine and rat pineal cell cultures, which is not due to cytotoxicity as judged by two different approaches, and 3) immunohistochemical evidence for the presence of nitric oxide synthase (NOS) (EC 1.14.23.-) in the intact bovine pineal gland and in cultured bovine pinealocytes. These data support the view that NOS is a component of the cGMP-generating system in mammalian pinealocytes. Although NO-donor molecules are also potent activators of cGMP accumulation, they may have other important actions in the pineal, namely the inhibition of adrenergic-stimulated melatonin synthesis. As SNP and SIN-1 exerted this inhibitory effect on cells regardless of whether they were stimulated by isoproterenol, forskolin or 8-Br-cAMP it would appear that NO-donors can act 'downstream' from the receptor/adenylate cyclase level.
TL;DR: The data suggest that mM concentrations of the pineal hormone are able to stimulate 5‐HT release from the Pinealocyte, while concentrations of melatonin increase extracellular 5‐ HT by inhibiting its reuptake in the adrenergic nerve endings.
Abstract: Melatonin has been proposed to exert some regulatory actions within the pineal gland itself. The present study examined the effect of melatonin on the release of serotonin (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA) from rat pineal glands by using an in vitro perifusion system. Melatonin induced a concentration-dependent stimulatory effect on 5-HT secretion from 10(-6) M to 10(-3) M. Maximal effects were obtained with melatonin 10(-3) M and concentrations lower than 10(-6) M were without effect. The secretion of 5-HIAA was inhibited by melatonin 10(-3) and 10(-4) M, but it was increased when pineals were incubated with 10(-5) and 10(-6) M of melatonin. The indoleamine secretion was also studied on peripherally denervated rat pineal glands. Basal output of 5-HT from these glands was increased when compared with those from control rats. In contrast, the secretion of 5-HIAA was strongly reduced after removal of the sympathetic input to the pineal gland. Melatonin 10(-3) M failed to stimulate 5-HT release from denervated pineal glands, although it inhibited 5-HIAA secretion. In contrast, melatonin 10(-5) M enhanced 5-HT release without altering 5-HIAA output. Fluoxetine, a 5-HT uptake inhibitor, produced similar effects than mM concentrations of melatonin on the indoleamine secretion from control pineal glands, but it had no effect on glands taken from peripherally denervated rats. These data suggest that mM concentrations of the pineal hormone are able to stimulate 5-HT release from the pinealocyte, while mM concentrations of melatonin increase extracellular 5-HT by inhibiting its reuptake in the adrenergic nerve endings. These findings are discussed in relation to the possible role of melatonin regulating the intra- and extracellular availability of 5-HT in the pineal gland and its significance as an autocrine factor.
TL;DR: The genesis of calcium concretions in aged rats was studied by means of transmission and scanning electron microscopy and the presence of sulfur indicates the existence of an organic matrix and it is proposed that genesis takes place in dark pinealocytes, which contain more calcium than light pinealocyte.
Abstract: The genesis of calcium concretions in aged rats was studied by means of transmission and scanning electron microscopy. The potassium pyroantimonate method, combined with X-ray microanalysis, allowed us to study the distribution of cations and calcium. Notable accumulations of calcium (associated with phosphorus) were localized in vesicles, vacuoles, lipid droplets, lipopigments, and mitochondria of dark pinealocytes. The results obtained in the present investigation suggest that these organelles are involved in the genesis of the concretions. The presence of sulfur indicates the existence of an organic matrix. We propose that genesis takes place in dark pinealocytes, which contain more calcium than light pinealocytes. Mineralization foci are some-times associated with cellular debris and enlarge by further apposition of material. Two types of concretions, as determined by electron microscopy and confirmed by electron diffraction, could be observed: the “amorphous” type with concentric layers and the crystalline type with needle-shaped crystals. Once formed, the concretions reach the extracellular space and the cell breaks down. Possible extracellular calcification is suggested in the extracellular calcium-rich floculent material. The mineralization process is interpreted as being an age-related phenomenon and mainly a consequence of the degeneration of pinealocytes.
TL;DR: Cat pinealocytes showed a nucleus with prominent nucleoli, a well developed Golgi apparatus, centrioles, granular endoplasmic reticulum, ribosomes, abundant microtubuli and enlarged mitochondria and their processes were characterized by the presence of abundant filaments.
Abstract: The ultrastructure of the pineal gland in the adult cat is described and compared with that of other mammals. Connective tissue spaces showed capillaries with nonfenestrated endothelia and numerous unmyelinated nerve fibers. In the proximal region of the gland, myelinated nerve fibers coming from the anterior commissure were also found. Cat pinealocytes showed a nucleus with prominent nucleoli, a well developed Golgi apparatus, centrioles, granular endoplasmic reticulum, ribosomes, abundant microtubuli and enlarged mitochondria. Pinealocytes showed several long processes with bulbous endings filled with clear vesicles and scarce "synaptic" ribbons. Pineal astrocytes and their processes were characterized by the presence of abundant filaments.
TL;DR: There is a significant increase in the number of dark pinealocytes there is a decrease in the total number of pinealo‐cytes in aged rats, which may explain the age‐related functional decline of the pineal gland activity.
Abstract: In the present study, we investigated the population of pinealocytes in the pineal gland of aging rats. Dark and light pinealocytes were analyzed as to their calcium content. Calcium localization was realized in dark and light cells by means of cytochemistry and X-ray microanalysis. Calcium was mainly localized in dark pinealocytes characterized by many ultrastructural signs of degeneration. The number of pinealocytes per square surface of aged rats (28 months) was com-pared to young ones (3–4 months). While there is a significant increase in the number of dark pinealocytes there is a decrease in the total number of pinealo-cytes in aged rats. This age-related loss of pinealocytes may explain the age-related functional decline of the pineal gland activity (e.g., the decrease of the nocturnal melatonin production).
TL;DR: The pineal gland is mainly sympathetic and pineal synthesis of melatonin and its levels in the circulation are under strict adrenergic control of serotoninN-acetyltransferase (NAT).
Abstract: 1. Innervation of the mammalian pineal gland is mainly sympathetic. Pineal synthesis of melatonin and its levels in the circulation are thought to be under strict adrenergic control of serotoninN-acetyltransferase (NAT). In addition, several putative pineal neurotransmitters modulate melatonin synthesis and secretion.
TL;DR: An immunohistochemical analysis of the cellular localization of AMPA-type glutamate receptor subunits in the pineal gland of the macaque showed that this subunit is present in interstitial cells and astrocyte-like cells located in the parenchyma of the distal part of the gland.
Abstract: A role for glutamate in signalling mechanisms in the mammalian pineal gland is suggested by the high concentration of the transmitter in pinealocytes and the expression of glutamate receptor subunits
TL;DR: Results indicate that extracellular melatonin may act as a paracrine or autocrine feedback signal and may be important for the illumination-dependent melatonin production.
TL;DR: The results suggest that the adrenergic-stimulated cAMP signal is subjected to the tonic inhibition by phosphoprotein phosphatase, and the camp signal is more sensitive to the regulation by phosphorylation than cGMP in rat pinealocytes.
Abstract: The role of phosphoprotein phosphatase in the regulation of adenosine 3',5'-cyclic monophosphate (cAMP) and guanosine 3',5'-cyclic monophosphate (cGMP) accumulation in rat pinealocytes was investigated using the three phosphatase inhibitors calyculin A, tautomycin, and okadaic acid. Calyculin A (0.1 microM) was found to enhance the isoproterenol- and norepinephrine-stimulated cAMP accumulation six- and threefold, respectively, whereas tautomycin and okadaic acid were less effective. The effect of calyculin A was rapid (within 5 min) and persisted in the presence of phosphodiesterase inhibition. However, in contrast to protein kinase C activation or intracellular calcium elevation, the phosphatase inhibitors were less effective in potentiating the cAMP response stimulated by forskolin or cholera toxin, and their effects were not blocked by calphostin C or N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide. The adrenergic-stimulated cGMP response was also less sensitive to the phosphatase inhibition. Therefore, our results suggest that 1) the adrenergic-stimulated cAMP signal is subjected to the tonic inhibition by phosphoprotein phosphatase; 2) phosphatase inhibitors enhance cAMP synthesis through their actions at the receptor level; and 3) the cAMP signal is more sensitive to the regulation by phosphorylation than cGMP in rat pinealocytes.
TL;DR: Results indicate that tyrosine kinase inhibitors increase pineal cGMP accumulation through inhibition of the metabolism of cG MP.
TL;DR: Profound morphological modifications occur during postnatal development of the rat pineal gland by using three cytoarchitectonic markers (S‐100, calbindin‐D28k, and calretinin) that belong to the calmodulin/troponin C calcium‐binding protein family to follow events from postnatal day 1 to 20.
Abstract: Profound morphological modifications occur during postnatal development of the rat pineal gland. We have immunohistochemically followed those events from postnatal day 1 to 20 by using three cytoarchitectonic markers (S-100, calbindin-D28k, and calretinin) that belong to the calmodulin/troponin C calcium-binding protein family. In the developing rat pineal, anticalbindin-D28k antibody labels three cell types: immature and mature astrocytes and perivascular type II pinealocytes. During development, calbindin-D28k positive cells migrate from the base of the pineal stalk into the superficial part of the pineal. Calbindin-D28k, usually used as a neuronal marker in the central nervous system, recognizes in rat pineal precursor astrocytes 5 days before S-100 and labels a subpopulation somewhat different from S-100 positive astrocytes. Calretinin immunoreactivity appeared in the postero-superior part of the pineal and was abundant until postnatal day 5, then its density dramatically felt to leave, after postnatal day 20, an occasional population of cells whose morphology is compatible with neuron-like cells.
TL;DR: Since MVs are the organelles in which transmitters are stored, these results suggest that NSF is involved in the MV- mediated exocytosis of transmitters from endocrine cells.
Abstract: N-ethylmaleimide-sensitive fusion protein (NSF), a protein necessary for vesicular docking and/or fusion, was detected immunohistochemically in pinealocytes. NSF was distributed similarly to synaptophysin and vacuolar-type H(+)-ATPase (V-ATPase), marker proteins for synaptic-like microvesicles (MVs) abundantly present in pinealocytes. A subcellular fractionation study indicated that .> 95% of NSF was present as a membrane-bound form and that some NSF was associated with MVs. Like neuronal NSF, the protein was not solubilized from membranes with either 2 mM Mg-ATP or 2% sodium carbonate, suggesting that NSF was tightly bound to the membranes. NSF was also detected in purified MVs from bovine posterior pituitaries. Since MVs are the organelles in which transmitters are stored, these results suggest that NSF is involved in the MV-mediated exocytosis of transmitters from endocrine cells.
TL;DR: Results indicate that in the pineal organ of the domestic fowl the ontogenetic expansion of NSE-immunoreactive pinealocytes is paralleled by a regressive afferent innervation, and the N SE-positive pinealocyte transform from a bipolar (columnar) type to a multipolar type during post-hatching development, which may reflect the development of a neurosecretory-like capacity of the organ.
Abstract: Immunohistochemistry for neuron-specific enolase (NSE) revealed that NSE is localized in both a limited number of pinealocytes and intrinsic afferent neurons in the pineal organ of the domestic fowl. Furthermore, a computer-assisted three-dimensional imaging technique allowed to clarify the reverse distributional pattern of both elements: NSE-positive pinealocytes displayed a dense distribution especially in the vesicular portion of the gland, whereas NSE-immunoreactive nerve cells were mainly found in the pineal stalk. The number of NSE-positive intrinsic neurons in the pineal organ of chickens decreased rapidly after hatching, with a concentration of these elements in the basal portion (stalk) of the pineal organ. On the other hand, immunoreactive pinealocytes increased remarkably in the end-vesicle of the organ with age, followed by a gradual expansion toward the proximal portion. Thus, the spectacular increase in NSE-positive pinealocytes and the progressive reduction of reactive neurons occurred in parallel during the course of post-hatching development. NSE-immunoreactive pinealocytes displayed morphological characteristics of bipolar elements, endowed with an apical protrusion into the pineal lumen and a short basal process at younger stages, whereas multipolar types of NSE-positive pinealocytes were predominantly found in the adult domestic fowl. These results indicate that in the pineal organ of the domestic fowl (1) the ontogenetic expansion of NSE-immunoreactive pinealocytes is paralleled by a regressive afferent innervation, (2) the NSE-positive pinealocytes transform from a bipolar (columnar) type to a multipolar type during post-hatching development, and (3) these ontogenetic changes in the NSE-immunoreactivity and morphology of pinealocytes may reflect the development of a neurosecretory-like capacity of the organ.
TL;DR: The pineal gland of the Mongolian gerbil is analyzed for the presence of rab3 proteins and the results lend further support to the hypothesis that mammalian pinealocytes are endowed with a population of SLMVs that serve secretory functions.
TL;DR: The hypothesis that NGF is actively secreted by PG and that nnc are the principal source of this neurotophin is supported and supports the hypothesis that PG neurotrophic effects are due to PG non‐neuronal cells that proliferate to confluency by 7 days in culture.
Abstract: The pineal gland (PG) is a source of several trophic factors. In this study, PG and superior cervical ganglia (SCG) from Sprague-Dawley neonates (1-day-old) were co-cultured to test the hypothesis that endogenous release of PG NGF (or an NGF-like cytokine) is sufficient to promote survival of SCG neurons. Neuronal density of SCG neurons was significantly enhanced when co-cultured with PG for 7 days compared to SCG cultured alone. SCG survival and neurite formation in PG co-cultures was less than in SCG treated with exogenous NGF (100 ng/ml). The neurotrophic effect of PG co-cultures was abolished when 1% anti-NGF was added to the medium. Co-cultures of SCG neurons with established 7-day PG cultures induced extensive SCG neurite formation within 24 hr compared to SCG co-cultured with 1-day PG cultures. This suggests that PG neurotrophic effects are due to PG non-neuronal cells (nnc) that proliferate to confluency by 7 days in culture. S-antigen-positive pinealocytes did not proliferate in culture. There was decreased SCG survival when neurons were seeded onto PG cultures that had been previously killed by drying, which suggests that the neurotrophic effects of nnc are not substrate-dependent. Immunocytochemical characterization of PG nnc revealed a heterogenous mixture of astrocytes, macrophage/microglia, and fibroblasts. These findings support the hypothesis that NGF is actively secreted by PG and that nnc are the principal source of this neurotophin.