Amount Of Serotonin Research Articles

Localization of a sodium-dependent high-affinity serotonin transporter and recruitment of exogenous serotonin by the cestodeHymenolepis diminuta: an autoradiographic and immunohistochemical study

Uptake of serotonin by tissues of intact and hemitransected Hymenolepis diminuta was studied by autoradiography and immunohistochemistry. Hemitransected worms were incubated in balanced saline containing 10 µM [3H]serotonin and washed extensively. The density of silver grains over the serotonin-immunoreactive longitudinal nerve cords and commissural rings, male reproductive system, tissues surrounding the genital pouch, and deep longitudinal muscles was significantly greater than that over the parenchyma. The presence of serotonin in spermatozoa suggested a role for this amine in spermatozoon activity. In contrast, uptake of 10 µM [3H]serotonin in sodium-free saline was significantly reduced compared with that in balanced saline in all tissues examined except the parenchyma. Analysis of the data revealed that the sodium-dependent high-affinity serotonin transport system is localized primarily in the serotonergic-like neurons of H. diminuta, which suggests possible recycling of neuronally released serotonin. Following incubation of intact worms in vitro for 12 h in 5 µM [3H]serotonin, the density of silver grains was significantly higher over the serotonin-immunoreactive nerves, elements of the male reproductive system, tissues surrounding the genital pouch, and deep longitudinal muscles than over the parenchyma. These results demonstrate recruitment of exogenous serotonin by intact H. diminuta and suggest sequestration and concentration by the serotonin-immunoreactive neurons via the sodium-dependent high-affinity transporter. These data further suggest that although H. diminuta can synthesize serotonin, it may obtain serotonin from the host. Nonetheless, the amount of serotonin recruited by H. diminuta from the host in vivo compared with that which they synthesize is not known.

Antioxidative properties of serotonin and the bactericidal function of polymorphonuclear phagocytes.

We investigated the antioxidative properties of platelet-released serotonin on the bactericidal function of polymorphonuclear neutrophils (PMN) since there is a surprising co-incidence of low blood serotonin and an increased rate of infections. The antioxidative properties of serotonin were demonstrated by its suppressive effects on phagocytosis-associated, luminol-enhanced chemiluminescence (CL). The bactericidal activity of PMN was determined by a microbiological assay using opsonized Staphylococcus aureus. Serotonin suppresses luminol-enhanced chemiluminescence in a dose-dependent manner indicating an interaction with reactive oxygen species, which are responsible for effective bacterial killing during the phagocytosis-associated "respiratory burst". The modulation of the bactericidal function of PMN by serotonin is complex and depends upon the amount of serotonin: at concentrations normally present at sites of tissue injury and consecutive thrombus formation (10(-6) to 10(-5) M) bacterial killing increases by about 50%. In contrast, at pharmacological concentrations (10(-3) to 10(-2) M) an adverse effect can be observed: the elimination of opsonized S. aureus is reduced by 30 to 90%. Exogenous serotonin is capable of modulating important biological functions of human PMN in vitro. At appropriate concentrations, the antibacterial defence improves significantly probably due to reduced autooxidation, whereas higher concentrations counteract an efficient bacterial killing.

Effect of p-chlorophenylalanine (PCPA) on sleep and monoamines content in the brain of a lizard species

Administration of PCPA, a specific inhibitor of serotonin synthesis, induced a significant decrease of total sleep time in the lizard Ctenosaura pectinata. This effect was exerted on both quiet sleep and active sleep, but it was more intense on active sleep. Reduction in the amount of active sleep was due to a decrease in the number of the episodes not in their mean duration, since this parameter increased significantly from 5.97 s, under control conditions, to 11.77 s, 10.66 s and 8.85 s at 24, 48 and 72 h after PCPA injection, respectively. Neurochemical analysis showed a significant decrease in the amount of serotonin in the analyzed brain stem structures 12 h after PCPA administration. The possible participation of serotonergic mechanisms in the regulation of reptilian sleep is discussed.

CNTF Induces Raphe Neuronal Precursors to Switch from a Serotonergic to a Cholinergic Phenotypein Vitro

Ciliary neurotrophic factor (CNTF) is a multifunctional cytokine that mediates survival and differentiation of neurons as well as many other cell types. In this study, CNTF and leukemia inhibitory factor (LIF) reduced the apparent number of primary serotonergic neurons in E14 raphe culture by 90% as determined by immunocytochemistry for serotonin (5HT). The reduction in 5HT cell number was not due to neuronal loss as removal of CNTF after 4 days in culture resulted in a partial restitution of the serotonergic phenotype. In the RN46A serotonergic cell line which is induced to become serotonergic by brain-derived neurotrophic factor (BDNF), the addition of CNTF suppressed tryptophan hydroxylase and 5HT synthesis and increased choline acetyl transferase (ChAT) expression by 6-fold and ChAT activity by 20- to 30-fold over 12 days. As with the primary neurons, removal and replacement of CNTF with BDNF after 4 days resulted in a partial restitution of 5HT expression. Moreover, other members of the CNTF-cytokine family that use gp130 and/or LIF receptor β as their signal transducing receptors—LIF, oncostatin M, interleukin 6, and interleukin 11—had similar effects on increasing ChAT activity and reducing 5HT expression in RN46A cells. Analysis of 5HT levels showed no significant difference in the amount of serotonin between wild-type and CNTFRα knockout mice at birth, suggesting that the potential to switch phenotype mediated through CNTFRα is a latent property of neuroepithelial precursors in the raphe nucleus.

Immunogold localization of serotonin within synaptic terminals in the rat mesencephalic trigeminal nucleus.

With the use of postembedding electron-microscopic immunogold cytochemistry, the vesicular distribution of serotonin within serotonergic synaptic terminals in the mesencephalic trigeminal nucleus was determined in order to obtain further insight into the mechanisms and functional significance of serotonin release to these jaw muscle spindle afferent neurons. Immunogold labelling was restricted to the previously described type I and type II terminals. The distribution of immunogold particles over the synaptic terminals indicated that serotonin was stored in small round or pleomorphic (RSV) vesicles and in large granular dense-cored (DSV) vesicles. The amount of serotonin present in the DSV vesicles, which are generally considered to contain colocalized neuropeptides, showed a high degree of variation. These DSV vesicles were usually located at some distance from the synaptic area of the terminals suggesting that they represent a nondirectly releasable vesicle pool. The serotonergic RSV vesicles were, in general, irregularly distributed over the terminals. However, in about 19% of the analyzed labelled synaptic terminals serotonergic RSV vesicles were clustered together near their release site at the synaptic cleft. These synaptic terminals may represent highly active serotonergic synapses with a high release frequency and a high reuptake level, resulting in a dense concentration of vesicles containing a high amount of serotonin near the synaptic cleft.

Isolation of serotonin from the accessory salivary glands of the marine snail Nucella lapillus

We report here the presence of a bioactive compound in the secretion of the accessory salivary glands (ASGs) of Nucella lapillus. We have purified the compound using HPLC and identified it as serotonin by mass spectrometry, UV spectroscopy, HPLC and capillary electrophoresis. Serotonin was not found in the secretions of the acinous salivary glands or the hypobranchial gland. The amount of serotonin in the secretion of the ASGs does not show seasonal or regional variation.

Serotonin in human tears.

It has been postulated that serotonergic receptors are present in the corneal epithelium and that their activation by serotonin released from subepithelial corneal nerves raises the level of cyclic AMP, which in turn stimulates active CI-secretion by the corneal epithelium. We looked for serotonin in tears because these bathe the corneal epithelium. Twenty-two normal subjects, 14 women and 8 men between 25 and 60 years of age (average 36.5 +/- 9 years), participated in this study. Twenty microliters of tears were collected in capillary tubes after trigeminal stimulation (pepper in the nose) and were immediately analysed using high-performance liquid chromatography with electrochemical detection. Serotonin was identified and measured in every subject. Its mean concentration was 2.74 +/- 1.99 ng/ml. No difference was found between women and men. Emotionally evoked tears were obtained from two subjects. The amount of serotonin in these samples was much higher than in the samples obtained by trigeminal stimulation. These results demonstrate for the first time that serotonin is present in human tears. The possible role of this serotonin in chloride transport in the corneal epithelium and the probable difference between "emotional" and stimulated tears warrants further investigation.

Mechanisms by which cancer chemotherapeutic drugs induce emesis.

Because the area postrema seems essential for chemotherapy-induced vomiting, both circulating and/or neurally mediated stimuli in this area could trigger the emetic response. In our laboratories results of cross-circulation and direct intracerebroventricular infusion experiments in dogs do not support a role for circulating substances. The large increases in serum vasopressin induced by cisplatin were not blocked by inhibitors of the angiotensin-converting enzyme. In the ferret inhibition of serotonin synthesis with p-chloro-phenylalanine, administration of selective antagonists of 5-hydroxy-tryptamine3 (5-HT3) receptors, or visceral deafferentation inhibited the emetic response evoked by cisplatin or high-dose cyclophosphamide. The results suggest that serotonin plays an important role and that peripheral neural mechanisms are involved in the emetic response. The strong antiemetic efficacy of selective 5-HT3 antagonists also has been confirmed in humans. In cancer patients high-dose cisplatin increased the plasma and urinary levels of 5-hydroxy-indoleacetic acid (5-HIAA), but did not affect platelet and free plasma serotonin. The changes in 5-HIAA levels paralleled the onset and development of vomiting. No evidence of serotonin depletion has been obtained after high-dose cisplatin. Dacarbazine, another strongly emetogenic agent, increased urinary 5-HIAA; however, only small increases in 5-HIAA were produced with low-dose cisplatin or cyclophosphamide-containing regimens. Thus, emetogenicity appears to be directly related to the ability of the cytotoxic agent to release serotonin. In humans, antiemetics such as ondansetron, metoclopramide, and dexamethasone did not effect high-dose cisplatin-induced increases in serotonin metabolism. Therefore, these antiemetics seem not to affect the amount of serotonin released. The mechanism by which chemotherapeutic drugs induce serotonin release is unknown; however, release may occur by direct cytotoxicity on the gastrointestinal mucosa, including the enterochromaffin cells. Delayed emesis appears to be mediated by 5-HT3-independent mechanisms. It is proposed that emesis that develops despite high-dose ondansetron (residual emesis) should be considered delayed emesis. Residual and delayed episodes of emesis have similar time courses, are characterized by very mild emetic episodes and poor response to 5-HT3 antagonists, and are not associated with increases in serotonin metabolism.

Serotonin plays a major role in serum-induced phospholipase C-mediated hydrolysis of phosphoinositides and DNA synthesis in vascular smooth muscle cells

Whole blood serum (WBS) rapidly induced the phospholipase C-mediated hydrolysis of phosphoinositides and subsequently stimulated DNA synthesis in cultured rabbit vascular smooth muscle cells (VSMCs). Ketanserin, a serotonin (S2) receptor antagonist, markedly inhibited the WBS-induced phospholipase C reaction and DNA synthesis. Serotonin by itself had a weak mitogenic activity for VSMCs, but this vasoconstrictor markedly stimulated the platelet-derived growth factor- and epidermal growth factor-induced DNA synthesis. The stimulatory effect of serotonin on the growth factor-induced DNA synthesis was inhibited by ketanserin. The amount of serotonin contained in WBS was sufficient to induce the phospholipase C reaction and stimulate the growth factor-induced DNA synthesis. These results indicate that serotonin plays a major role in the WBS-induced phospholipase C-mediated hydrolysis of phosphoinositides and DNA synthesis in rabbit VSMCs and suggest that serotonin may act as an important growth regulator for VSMCs in addition to acting as a vasoconstrictor.

Subcellular composition of platelet density subpopulations prepared using continuous Percoll gradients

Normal human platelets have been separated by density on continuous Percoll gradients and the subcellular composition of platelets of different density has been analysed. The number and concentration of dense granules increased significantly with platelet density, as did the concentrations of the dense granule constituents calcium and serotonin. The amount of serotonin per granule in the low density (LD) platelets was only two-thirds of the corresponding amount in the high density (HD) platelets. Platelets of all densities were able to sequester exogenous serotonin and release it in response to thrombin stimulation with similar efficiencies. The concentrations of the alpha-granule constituents von Willebrand factor and beta-thromboglobulin increased significantly with platelet density but the concentrations of the lysosomal enzyme beta-N-acetylglucosaminidase and total sialic acid did not differ significantly in the density subpopulations. The concentrations of the cytosolic enzymes lactate dehydrogenase and glucose-6-phosphate dehydrogenase were slightly higher in the LD population than in the other density subpopulations. The concentration of glycogen showed a marked positive relationship with platelet density and calculations suggested that glycogen was an important determinant of platelet density heterogeneity. The findings of the present study are compatible with recent suggestions that LD platelets may be slightly younger than HD platelets in normal human subjects.

Serotoninergic neurons in the retina of Xenopus laevis: selective staining, identification, development, and content.

Uptake of 3H-serotonin followed by autoradiography, and uptake of the serotonin analog 5,7-dihydroxytryptamine (5,7-DHT), with subsequent staining, were each used to define a unique set of neurons in the retina of the African clawed frog, Xenopus laevis. Both techniques demonstrated the same population of neurons, on the basis of perikaryal size, shape, and position within the retina. Two classes of amacrine cells accumulated 5,7-DHT at the proximal (vitread) margin of the inner nuclear layer; the two classes were distinguished by the size of their perikarya. Two similar populations of cells, observed in the ganglion cell layer with lower frequency, may represent "displaced" counterparts of these two amacrine cell types. A class of bipolar cells whose perikarya were located in middle-to-distal regions of the inner nuclear layer also accumulated 5,7-DHT and 3H-serotonin. Processes of these cells contributed to a dense plexus of fine fibers that appeared evenly distributed throughout the inner plexiform layer. 3H-Serotonin-accumulating cells first appeared in the developing retina at stage 35/36, a time immediately after retinal stratification but before elaboration of either plexiform layer. Electron microscopic analysis permitted an identification of 3H-serotonin-accumulating terminals in the inner plexiform layer. Serotonin-labeled terminals containing conventional contacts, suggestive of amacrine cells, were presynaptic to unidentified processes and postsynaptic to bipolar cells. Labeled terminals containing ribbon contacts, indicative of bipolar cells, were postsynaptic to amacrine cells. The amount of serotonin contained in isolated retinas was 15 pmol/mg protein as measured by HPLC with electrochemical detection. We attempted to stimulate the release of accumulated 3H-serotonin from mature retinas by increasing the K+-concentration in the bathing medium. Although preloaded glycine is readily released from 14C-glycine-accumulating neurons, from the same retinas there was no calcium-dependent, K+-stimulated release of 3H-serotonin. This finding suggests that serotonin and glycine are processed differently by retinal neurons, the consequence of which results in differing responses to 40 mM K+.

Effects of Anoxia on Serotonin Metabolism in Crucian Carp Brain

ABSTRACT In the brain, oxygen is required for both the synthesis and the degradation of monoamine transmitters, so monoaminergic systems can be expected to be strongly affected by anoxia. However, crucian carp (Carassius carassius L.) may survive anoxia for many days or even weeks. In the present study, crucian carp were exposed to anoxia for 22, 76, and 160 h at 8°C. All survived and were found to excrete ethanol at a constant rate. The brain concentrations of serotonin and its two main metabolites, 5-hydroxyindole-3-acetic acid (5-HIAA) and 5-hydroxytryptophol, were analysed after each experiment. In a preliminary experiment, it was found that the brain of the crucian carp contained about the same amount of serotonin and 5-HIAA as two species less tolerant to anoxia - the common carp and the rainbow trout. The levels of the serotonin metabolites decreased drastically (by 80–90%) during anoxia, whereas serotonin levels were only slightly reduced (by 15 % or less). These results suggest a complete or nearly complete stop in serotonin metabolism during anoxia.

Induced variation of serotonin in Drosophila melanogaster and its relation to learning performance.

In insects the level of serotonin can be modified through the alimentary tract. Drosophila fed on chemically defined media to which has been added a synthesis inhibitor or an immediate precursor of this neurotransmitter have their serotonin level decreased or increased, respectively. (A 4- to 5-day treatment is optimal). Such treated flies, if tested in a procedure of Pavlovian conditioning, have different learning performances. With the same duration of starvation, nearly the same threshold for sucrose solutions, but different amounts of serotonin, the flies (blindly trained) showed conditioned inhibition of the tarsal (or proboscis-extension) reflex according to the amount of serotonin. The highest level of this biogenic amine gave the best performance.

Binding of Aggregated Immunoglobulins to the Human Platelet Fc Receptors: A Mechanism of Platelet to Platelet Bridging

Aggregated immunoglobulins react with human platelets by occupying the Fc receptors present on their surface, inducing aggregation and the release reaction. We studied the effect of heat aggregated gammaglobulins (HAGG) on ADP-induced aggregation of platelets. We used the minimum concentration of ADP required to induce a reversible aggregation of platelets without any substantial amount of serotonin (14C-5HT) release. EDTA (5 mM) added at the peak of platelet aggregation resulted in rapid deaggregation of these platelets. However, incubation of platelets with HAGG at a dose that did not by itself induce any aggregation or release reaction, followed by ADP addition resulted in an irreversible platelet aggregation of greater magnitude accompanied by a substantial release of 14C-5HT. The addition of EDTA at the peak of platelet aggregation failed to deaggregate these platelets. To determine whether the augmented aggregation response and the inhibition of deaggregation was due to HAGG or a consequence of platelet release products, we used thrombin-degranulated platelets. The augmented aggregation response and the inhibition of deaggregation due to HAGG and ADP could be demonstrated using these platelets. To confirm that the binding of HAGG to the platelet Fc receptors was responsible for these observations, we incubated platelets with an excess of Fc fragments of IgG prior to the addition of HAGG and ADP. This abolished the aggregation response observed previously. From this study we conclude that interplatelet bridging by HAGG renders the platelets hyperaggregable and appears to be a mechanism involved in maintaining platelet aggregates.

Novel Flushing Provoked by Volatile Anesthetics in <italic>Mastomys natalensis</italic> Bearing a Transplantable Substrain of Gastric Carcinoid That Predominantly Secretes Serotonin<xref ref-type="fn" rid="FN2">2</xref>

A tumor substrain secreting a large amount of serotonin [5-hydroxytryptamine (5-HT); CAS: 50-67-9; 3-(2-amino-ethyl)indol-5-ol] and a minute amount of histamine (CAS: 51-45-6) has been isolated from the previously established strain of transplantable gastric carcinoid of Mastomys (Praomys) natalensis secreting both histamine and 5-HT. Mastomys bearing a large growing transplant and excreting a large amount of 5-hydroxy-indoleacetic acid [(5-HIAA) CAS: 54-16-0] were associated often with reddening of the nose, lower lip, auricles, hands, and feet. Soon after the animals were anesthetized by ether or other volatile anesthetics, the tinges of red of the above-mentioned exposed parts abruptly turned bright red and rapidly spread over the neck, upper chest, and epigastric area. The reddening was transient, lasting 1.5-5 minutes, thereby fulfilling the criteria of flushing. The severity of ether-provoked flushing in tumor-bearing Mastomys paralleled the urinary excretion levels of 5-HIAA. The ether-provoked flushing was prevented completely by sc injection of either ketanserin (150 micrograms) or somatostatin (20 micrograms). The same ether-provoked flushing as found in tumor-bearing Mastomys could be reproduced in normal ones by constant infusion of 20 mg 5-HT/kg/24 hours (i.e., doses comparable to those released from a transplanted tumor) through an osmotic minipump implanted subcutaneously.

Serotonin in neuroblastoma × glioma NG108-15 hybrid cells

Mouse neuroblastoma × rat glioma NG108-15 hybrid cells contain a considerable amount of serotonin, and possess small but significant tryptophan hydroxylase activity. The results suggest that NG108-15 hybrid cells are serotonergic, in addition to the known cholinergic property.

Platelet Shape Change and Cytoskeletal Assembly: Effects of pH and Monovalent Cation lonophores

The monovalent cation ionophores monensin and nigericin cause platelet shape change at a rate of approximately 1/20 of that caused by ADP. The effect of monensin was studied further. Shape change caused by monensin is pH dependent, increasing in rate as extracellular pH increases. Monensin induced shape change is not blocked by 30 microM cinanserin which completely inhibits serotonin induced shape change. Also, the amount of serotonin secreted by monensin treated platelets is below the threshold required to induce shape change. 100 microM ATP which inhibits ADP induced shape change does not affect monensin induced shape change. Amiloride, a sodium transport blocker, inhibits both the rate of ADP induced shape change and platelet spreading on poly-lysine coated glass. Amorphous platelet cytoskeletons isolated from resting platelets at pH 6.8 with Mg++ but not Ca++ can be transformed into filament bundles by subsequent incubation at pH 7.6. We conclude that platelet shape change is at least in part triggered by changes in cellular Na+ and pH.

Serotonin-containing neurones in vertebrate retinas.

It has been established by a combination of HPLC and electrochemical detection that frog, lizard, goldfish, rabbit, and bovine retinas contain both dopamine and serotonin. Immunohistological and immunoradiographical methods show that serotonin is localised in amacrine perikarya and processes situated in the inner plexiform layers of frog, lizard, and goldfish retinas. The amount of serotonin in the mammalian retina appears to be too low for detection in neurones. The serotonin in the bovine retina is located mainly in the inner nuclear and plexiform layers, suggesting that the amine is present in the same types of cells as found for frog, lizard, and goldfish retinas. Retinas incubated in [3H]serotonin showed that radioactivity is associated with processes in the inner plexiform layer and amacrine perikarya. These results suggest that the neuronal elements that contain endogenous serotonin also have the capacity to accumulate exogenous amine and are consistent with the opinion that serotonin has a neuronal function in retinas of a variety of vertebrates.

Contrast media induced serotonin release in human blood.

Several contrast media were tested for their ability to release serotonin from whole blood. Renografin-60, Hypaque M-75, Vascoray and Conray were incubated with human blood at 37 degrees C for 60 min. Platelet-poor plasma was analysed for serotonin using a high performance liquid chromatographic technique. Seven non-ionic contrast media were similarly incubated and serotonin levels determined. Ionic contrast media induced a greater release of serotonin compared with non-ionic ones. The amount of serotonin was not proportional to iodine concentration in blood. These results suggest that hypertonic solutions of contrast media produce a physical injury to platelets causing them to release their granular contents.

Tissues as receptors of photoperiodic monochromatic light and the regulation of serotonin metabolism

Abstract Photoperiods are recognized to induce a variety of physiological events in organisms, in particular, several daily or circadian rhythms. Hormones are secreted in daily rhythm patterns and provide regulation of other rhythmic physiological events. Serotonin is understood by several investigators to be a hormone that is produced in response to photoperiods, and its pattern of secretion is circadian in output, e.g. under long term tissue culture conditions (previous study). This study shows that monochromatic lights in photoperiods cause circadian outputs of serotonin. Yellow light (578 nm) produced the serotonin secretion pattern most nearly similar to white (control) light. The highest levels of serotonin were seen in the early evening which was maintained throughout the night. Each end of the visible spectrum produced the least overall amount of serotonin in time and the middle of the spectrum the greatest amount. Notable exceptions to this general trend were the effects of ultra violet and viole...