Serotoninergic Cells Research Articles

QUERCETIN SUPPLEMENTATION PREVENTS CHANGES IN THE SEROTONIN AND CASPASE-3 IMMUNOREACTIVE CELLS OF THE JEJUNUM OF DIABETIC RATS.

Serotonin (5-HT) is present in the epithelial enterochromaffin cells (EC), mast cells of the lamina propria and enteric neurons. The 5-HT is involved in regulating motility, secretion, gut sensation, immune system and inflammation. Evaluate the effects of diabetes and quercetin supplementation on serotoninergic cells and its cell loss by apoptosis in jejunal mucosa of streptozotocin-induced diabetic rats (STZ-rats). Twenty-four male Wistar rats were divided into four groups: normoglycemic (C), normoglycemic supplemented with 40 mg/day quercetin (Q), diabetic (D) and diabetic supplemented with 40 mg/day quercetin (DQ). After 120 days, the jejunum was collected and fixated in Zamboni's solution for 18 h. After obtaining cryosections, immunohistochemistry was performed to label 5-HT and caspase-3. Quantification of 5-HT and caspase-3 immunoreactive (IR) cells in the lamina propria, villi and crypts were performed. The diabetic condition displayed an increase of the number of 5-HT-IR cells in villi and crypts, while decreased number of these cells was observed in lamina propria in the jejunum of STZ-rats. In the diabetic animals, an increased density of apoptotic cells in epithelial villi and crypts of the jejunum was observed, whereas a decreased number of caspase-3-IR cells was observed in lamina propria. Possibly, quercetin supplementation slightly suppressed the apoptosis phenomena in the epithelial villi and crypts of the STZ-rats, however the opposite effect was observed on the 5-HT-IR cells of the lamina propria. Quercetin supplementation on healthy animals promoted few changes of serotoninergic function and apoptotic stimuli. These results suggest that quercetin supplementation mostly improved the serotonergic function affected by diabetes maybe due to antioxidant and anti-inflammatory properties of quercetin.

Pain inhibition through transplantation of fetal neuronal progenitors into the injured spinal cord in rats.

Neuropathic pain after spinal cord injury (SCI) is a complex condition that responds poorly to usual treatments. Cell transplantation represents a promising therapy; nevertheless, the ideal cell type in terms of neurogenic potential and effectiveness against pain remains largely controversial. Here, we evaluated the ability of fetal neural stem cells (fNSC) to relieve chronic pain and, secondarily, their effects on motor recovery. Adult Wistar rats with traumatic SCI were treated, 10 days after injury, with intra-spinal injections of culture medium (sham) or fNSCs extracted from telencephalic vesicles (TV group) or the ventral medulla (VM group) of E/14 embryos. Sensory (von Frey filaments and hot plate) and motor (the Basso, Beattie, Bresnahan locomotor rating scale and inclined plane test) assessments were performed during 8 weeks. Thereafter, spinal cords were processed for immunofluorescence and transplanted cells were quantified by stereology. The results showed improvement of thermal hyperalgesia in the TV and VM groups at 4 and 5 weeks after transplantation, respectively. Moreover, mechanical allodynia improved in both the TV and VM groups at 8 weeks. No significant motor recovery was observed in the TV or VM groups compared with sham. Stereological analyses showed that ~70% of TV and VM cells differentiated into NeuN+ neurons, with a high proportion of enkephalinergic and GABAergic cells in the TV group and enkephalinergic and serotoninergic cells in the VM group. Our study suggests that neuronal precursors from TV and VM, once implanted into the injured spinal cord, maturate into different neuronal subtypes, mainly GABAergic, serotoninergic, and enkephalinergic, and all subtypes alleviate pain, despite no significant motor recovery. The study was approved by the Animal Ethics Committee of the Medical School of the University of São Paulo (protocol number 033/14) on March 4, 2016.

Organization of the Orexin/Hypocretin System in the Brain of Holostean Fishes: Assessment of Possible Relationships with Monoamines and Neuropeptide Y

Holosteans form a small group of actinopterygian fishes considered the sister group of teleosts. Despite this proximity to the biggest group of vertebrates, relatively few studies have been conducted to investigate the organization of the central nervous system of this group of fishes. In this study, the neuroanatomical distribution of orexin/hypocretin-like immunoreactive (OX-ir) cell bodies and fibers was analyzed in the brain of 3 representative species of the 2 orders of extant holosteans, the spotted gar Lepisosteus oculatus, the Florida gar Lepisosteus platyrhincus, and the bowfin Amia calva. Antibodies against orexin-A (OXA) and orexin-B (OXB) were used, which labeled the same cells and fibers throughout the brain. In addition, double immunohistofluorescence was performed for the simultaneous detection of OXA and OXB with tyrosine hydroxylase, serotonin, and neuropeptide Y (NPY), in an attempt to localize the orexinergic structures precisely and study the possible interactions between these neuroactive substances. The pattern of distribution of OX-ir cells in the 3 species was largely similar, showing labeled cells in the preoptic area (POA), and the tuberal and retrotuberal hypothalamic regions, with only subtle differences between species in the density of labeled cells. OX-ir fibers were found in all main brain subdivisions of the 3 species, mostly in the ventral subpallial areas, POA, hypothalamus, posterior tubercle, thalamus, and mesencephalic tectum. Different densities of orexinergic fibers were observed in relation to catecholaminergic and serotoninergic cell groups, as well as an absence of colocalization between orexins and NPY in the same hypothalamic neurons. The comparison of these results with those obtained in other vertebrates highlights a constant pattern of distribution of this system of neurotransmission among different groups of actinopterygian fishes, especially in teleosts. Conserved features shared by all vertebrates studied were also observed, such as the presence of OX-ir cells in the basal hypothalamus, reflecting the preserved functions of these neuropeptides over the course of evolution.

Effect of Gender on Chronic Intermittent Hypoxic Fosb Expression in Cardiorespiratory-Related Brain Structures in Mice.

We aimed to delineate sex-based differences in neuroplasticity that may be associated with previously reported sex-based differences in physiological alterations caused by repetitive succession of hypoxemia-reoxygenation encountered during obstructive sleep apnea (OSA). We examined long-term changes in the activity of brainstem and diencephalic cardiorespiratory neuronal populations induced by chronic intermittent hypoxia (CIH) in male and female mice by analyzing Fosb expression. Whereas the overall baseline and CIH-induced Fosb expression in females was higher than in males, possibly reflecting different neuroplastic dynamics, in contrast, structures responded to CIH by Fosb upregulation in males only. There was a sex-based difference at the level of the rostral ventrolateral reticular nucleus of the medulla, with an increase in the number of FOSB/ΔFOSB-positive cells induced by CIH in males but not females. This structure contains neurons that generate the sympathetic tone and which are involved in CIH-induced sustained hypertension during waking hours. We suggest that the sex-based difference in neuroplasticity of this structure contributes to the reported sex-based difference in CIH-induced hypertension. Moreover, we highlighted a sex-based dimorphic phenomenon in serotoninergic systems induced by CIH, with increased serotoninergic immunoreactivity in the hypoglossal nucleus and a decreased number of serotoninergic cells in the dorsal raphe nucleus in male but not female mice. We suggest that this dimorphism in the neuroplasticity of serotoninergic systems predisposes males to a greater alteration of neuronal control of the upper respiratory tract associated with the greater collapsibility of upper airways described in male OSA subjects.

Protein Kinases Alter the Allosteric Modulation of the Serotonin Transporter In Vivo and In Vitro.

Studies using S- and R-enantiomers of the SSRI citalopram have shown that R-citalopram exerts an antagonistic effect on the efficacy of the antidepressant S-citalopram (escitalopram) through an interaction at an allosteric modulator site on the serotonin transporter (SERT). Here, we show that protein kinase signaling systems are involved in the allosteric modulation of the SERT invivo and invitro. We assessed the effects of nonspecific protein kinase inhibitor staurosporine in the action of escitalopram and/or R-citalopram using electrophysiological and behavioral assays in rats and cell surface SERT expression measures in serotoninergic cells. Acute administration of R-citalopram counteracted the escitalopram-induced suppression of the serotonin (5-HT) neuronal firing activity and increase of the head twitches number following L-5-hydroxytryptophan injection. Importantly, these counteracting effects of R-citalopram were abolished by prior systemic administration of staurosporine. Interestingly, the preventing effect of staurosporine on 5-HT neuronal firing activity was abolished by direct activation of protein kinase C with phorbol 12-myristate 13-acetate. Finally, invitro, quantification of the amount of cell surface-expressed SERT molecules revealed that R-citalopram prevented escitalopram-induced SERT internalization that was completely altered by staurosporine. Taken together, these results highlight for the first time an involvement of protein kinases in the allosteric modulation of SERT function.

Supplementation with L-Glutamine and L-Alanyl-L-Glutamine Changes Biochemical Parameters and Jejunum Morphophysiology in Type 1 Diabetic Wistar Rats.

We evaluated the effects of the supplementation with L-glutamine and glutamine dipeptide (GDP) on biochemical and morphophysiological parameters in streptozotocin-diabetic rats. For this purpose, thirty animals were distributed into six groups treated orally (gavage) during thirty days: non diabetic rats (Control) + saline, diabetic + saline; Control + L-glutamine (248 mg/kg), Diabetic + L-glutamine (248 mg/kg), Control + GDP (400 mg/kg), Diabetic + GDP (400 mg/kg). Diabetes was induced by an intravenous injection of streptozotocin (60 mg/kg) and confirmed by fasting glucose ≥ 200 mg/dL. Physiological parameters, i.e., body mass, food intake, blood glucose, water intake, urine and faeces were evaluated during supplementation. After the period of supplementation, the animals were euthanized. The blood was collected for biochemical assays (fructosamine, transaminases, lipid profile, total protein, urea, ammonia). Moreover, the jejunum was excised and stored for morphophysiological assays (intestinal enzyme activity, intestinal wall morphology, crypt proliferative index, number of serotoninergic cells from the mucosa, and vipergic neurons from the submucosal tunica). The physiological parameters, protein metabolism and intestinal enzyme activity did not change with the supplementation with L-glutamine or GDP. In diabetic animals, transaminases and fructosamine improved with L-glutamine and GDP supplementations, while the lipid profile improved with L-glutamine. Furthermore, both forms of supplementation promoted changes in jejunal tunicas and wall morphometry of control and diabetic groups, but only L-glutamine promoted maintenance of serotoninergic cells and vipergic neurons populations. On the other hand, control animals showed changes that may indicate negative effects of L-glutamine. Thus, the supplementation with L-glutamine was more efficient for maintaining intestinal morphophysiology and the supplementation with GDP was more efficient to the organism as a whole. Thus, we can conclude that local differences in absorption and metabolism could explain the differences between the supplementation with L-glutamine or GDP.

Food restriction beginning at lactation interferes with the cellular dynamics of the mucosa and colonic myenteric innervation in adult rats.

The effects of food restriction (FR) on the morphoquantitative aspects of the wall and myenteric neurons of the proximal colon in adult rats were analysed. FR was imposed by duplication of the experimental brood size in relation to the control brood during lactation. The FR group received a 50% reduction of food from weaning until 90 days of age. Samples of the colon underwent histological processing to morphometrically analyze the crypts, muscularis mucosae, tunica mucosa, and muscularis externa. We determined the number of goblet cells and serotoninergic enteroendocrine cells, and morphoquantitatively studied the myenteric neuronal population. FR caused hypertrophy in the tunica mucosa, increase in crypt depth and in the muscular layer of the mucosa, a decrease in the thickness of the tunica muscularis and in the number of goblet cells and an increase in serotoninergic cells. A higher neuronal density in the ganglia and a reduction of the cell profile area were observed in the FR group. FR imposed since lactation led to hypertrophy of the tunica mucosa, a reduction of neutral mucin production, atrophy of the tunica muscularis, and an increase in the survival neuronal in adult rats, attributable to an increase in the number of serotoninergic enteroendocrine cells in mucosa.

Galanin receptor/Neuropeptide Y receptor interactions in the dorsal raphe nucleus of the rat

The aim of this study was to evaluate by quantitative receptor autoradiography the interactions between Neuropeptide Y Y1 (NPY Y1) and Galanin (GAL) receptors in the dorsal raphe nucleus (DRN) where both GAL receptors and NPY Y1 receptors exist. The ability of the GAL receptor antagonist M35 to block the GAL action was also evaluated. Double immunocytochemical staining of 5-hydroxytryptmine and c-Fos and stereology techniques were used to study the specific cell activation in the DRN after the intracerebroventricular coinjections of GAL and the NPY Y1/Y5 agonist [ 125I] Leu 31,Pro 34PYY. GAL (0.3 nM) decreases [ 125I] Leu 31,Pro 34PYY binding in the DRN by 48% ( p < 0.01) as shown by quantitative receptor autoradiography. This effect was reversed with the GAL receptor antagonist M35. Intracerebroventricular coinjections of NPY Y1/Y5 agonist and GAL reduced the c-Fos expression in the serotoninergic cells induced by the NPY Y1/Y5 agonist in DRN. These results indicate the existence of antagonistic interactions between GAL receptors and NPY Y1 receptors in the DRN that may be of relevance in mood disorders.

Effects of chronic lead intoxication on rat serotoninergic system and anxiety behavior

Chronic lead exposure has been shown to produce behavioral disturbances in human and animal models. These disturbances are associated with alterations in monoaminergic neurotransmission in the central nervous system (CNS), some of which have been attributed to serotonin (5-HT). This study was undertaken to investigate the chronic effects of lead exposure on the serotoninergic system in the dorsal raphe nucleus (DRN) and the consequences of its toxicity on rat behavior. Adult male Wistar rats were chronically exposed for 3 months to 0.5% lead acetate in drinking water. The serotoninergic system was evaluated using immunohistochemistry and the anxiety behavior was assessed by the light/dark box test. The results show that chronic lead exposure induces a significant increase of blood and brain lead levels in treated rats compared with controls. The density of the immunoreactive serotoninergic cell bodies was significantly higher in treated rats in all parts of the DRN. Assessment of animal behavior using the light/dark box test showed that lead-treated rats spent significantly more time in the light chamber compared with controls ( P = 0.001). These findings suggest that lead exposure may possibly induce increased anxiety as a consequence of changes in neuronal 5-HT content in the DRN.

Distribution of Orexin/Hypocretin Immunoreactivity in the Brain of the Lungfishes Protopterus dolloi and Neoceratodus forsteri

Lungfishes are currently considered the closest living relatives of tetrapods and represent an interesting group for the study of evolutionary traits in the transition from fishes to tetrapods. The brains of lungfishes have received little attention in comparative studies probably due to the difficulty of obtaining these unique animals. In the present study the distribution of orexin (hypocretin)-like immunoreactivity was studied in the brain of the African lungfish Protopterus dolloi and the Australian lungfish Neoceratodus forsteri by using antibodies directed against the mammalian orexin-A and orexin-B peptides. Simultaneous detection of orexins and tyrosine hydroxylase or serotonin was used to assess the precise location of the orexins in the brain and to evaluate the possible influence of the orexin system on the monoaminergic cell groups. Although some differences were noted, a common pattern for the distribution of orexins in the two lungfishes studied was observed. In both species, most immunoreactive neurons were observed in the suprachiasmatic nucleus and dorsal hypothalamus. Only in Neoceratodus, however, were important cell populations found in the preoptic area and infundibular hypothalamus, whereas small numbers of faintly reactive neurons were present in the lateral septum and ventral striatum. Fiber labeling was widely distributed in all main brain subdivisions, but was more abundant in regions such as the septum, preoptic area, suprachiasmatic nucleus, lateral hypothalamic area, thalamus, pretectum and tegmentum. Less conspicuous was the innervation of the pallial regions, habenula, optic tectum, rhombencephalic reticular formation and spinal cord. Orexinergic innervation was found in close contact with dopaminergic, noradrenergic and serotoninergic cell groups, homologous to the substantia nigra in the midbrain tegmentum, the locus coeruleus, the nucleus of the solitary tract and the raphe nuclei. Although unique features have been found for lungfishes, the location of orexin immunoreactive elements is largely consistent with that recently reported following a similar approach in amphibians and amniotes, suggesting that the general organization of this peptidergic system occurred in the common ancestor of lungfishes and tetrapods.

Effects of Serotonin on Acid Secretion in Isolated Rat Stomach: the Role of 5-HT3 Receptors

Serotonin (5-hydroxytryptamin; 5-HT) content has been measured using high-performance liquid chromatography with electrochemical detection (HPLC-ECD). The distributions of 5-HT-containing cells and 5-HT3 receptors have been determined with specific antibodies against 5-HT and 5-HT3 receptors, respectively. The effect of serotonin on acid secretion has been studied using an isolated rat stomach model. It has been shown that 5-HT concentrations in the fundus, mucosal layers of the corpus, remaining layer of the corpus and antrum are approximately 152, 498, 1494 and 972 nmol/mg protein, respectively. The distribution of 5-HT-containing cells is concentrated in the enteric plexus and enterochromaffin (EC) cells in the deep mucosal layer. Immunoreactivity to 5-HT3 receptors is localized in numerous neurons of the myenteric and submucosal plexus and concentrated in the neuronal plasma membrane, submucosa, endocrine cells and lamina propria. In the present study, the effect of 5-HT on gastric acid secretion was investigated on an everted preparation of isolated rat stomach. 5-HT at 1-100 microM reduced acid secretion stimulated by oxotremorine while 10 microM 5-HT did not modify the basal secretion of gastric acid. We further showed that 10 microM 5-HT reduced acid secretion and pepsin output stimulated by oxotremorine, histamine and pentagastrin; among the 5-HT receptors agonists tested, 2-methyl-5-HT (1-10 microM), a 5-HT3 receptor agonist, inhibited oxotremorine-, histamine- and pentagastrin-stimulated acid secretions, and this inhibitory effect was blocked by 1 microM MDL 72222, a specific 5-HT3 receptor antagonist. These results suggest that 5-HT is released from serotoninergic neurons, their processes and EC cells. The effect of 5-HT mediated by 5-HT3 receptors involves distinct neuronal and non-neuronal pathways which modulate gastric acid secretion.

Complex regional influence of photoperiod on the nycthemeral functioning of the dorsal and median raphé serotoninergic system in the Syrian hamster

The Syrian hamster (Mesocricetus auratus) is a widely used species for the study of biological clock synchronization and photoperiodism. The serotoninergic system arising from the median (MnR) and the dorsal raphé (DR) is a major actor in circadian clock synchronization. This serotoninergic system is also associated with functions and behaviours influenced by seasonal changes. The aim of the present study was to assess the influence of photoperiod on the daily functioning of the MnR and DR serotoninergic system. The morphology of both raphé nuclei was analysed in hamsters kept under long and short photoperiod by immunocytochemical detection of two markers of the serotoninergic system, serotonin and tryptophan hydroxylase (TPH, the rate-limiting enzyme of serotonin synthesis). The morphological analysis revealed a fairly complex morphological organization of the DR and MnR along their caudo-rostral extent. This morphological organisation was similar in the two photoperiods. However, quantification of several markers of serotonin (5-HT) synthesis in the DR and MnR revealed a time-dependent functioning of serotoninergic cells that was locally influenced by photoperiod. In particular, the significant daily variations of tph2-mRNA and TPH levels in the rostral MnR, and of the 5-HT/5-HIAA (5-hydroxyindoleacetic acid) ratio within the suprachiasmatic nuclei, were abolished under short photoperiod. The results are discussed with regard to the known physiological role of the serotoninergic system on the biological clock.

Immunohistochemical localization of orexins (hypocretins) in the brain of reptiles and its relation to monoaminergic systems

With the aim of gaining more insight into the evolution of the orexinergic systems in the brain of vertebrates we have conducted a comparative analysis of the distribution of orexin-immunoreactive cell bodies and fibers in two reptiles, the lizard Gekko gecko and the turtle Pseudemys scripta elegans. In both species most immunoreactive neurons were found in the periventricular hypothalamic nucleus and in the infundibular hypothalamus. Only in the gecko, orexinergic cell bodies were present in the dorsolateral hypothalamic nucleus and the periventricular preoptic nucleus. Fiber labeling was observed in all main brain subdivisions but was more abundant in regions such as the septum, preoptic area, suprachiasmatic nucleus, lateral hypothalamic area and median eminence. Less conspicuous was the innervation of the olfactory bulbs, pallial regions, habenula, dorsomedial and dorsolateral thalamic nuclei, torus semicircularis and spinal cord. Double immunohistofluorescence techniques were applied for the simultaneous detection of the orexinergic systems and the catecholaminergic or serotoninergic systems in the brain of reptiles. Actual colocalization of orexins and catecholamines or serotonin in the same neurons was not observed. However, orexinergic innervation was found in dopaminergic, noradrenergic and serotoninergic cell groups, such as the substantia nigra and ventral tegmental area in the midbrain tegmentum, the locus coeruleus, the nucleus of the solitary tract and the raphe nuclei. The comparison of the distribution of orexin-immunoreactive neurons and fibers found in reptiles with those reported for other vertebrates reveals a strong resemblance but also notable variations. In addition, the relation between the orexinergic and monoaminergic systems observed in the brain of reptiles seems to be a shared feature among vertebrates.

Functional correlates for 5-HT 1A receptors in maternally deprived rats displaying anxiety and depression-like behaviors

Maternal separation is known to induce long-term changes in neuroendocrine and emotional responsiveness to stress in a large variety of models. We examined an animal model of early deprivation in Sprague–Dawley rats consisting of separating litters from their mothers and littermates 3 h daily during postnatal days 2 to 15. In adulthood, maternally deprived rats in comparison with non-deprived controls exhibited an increase in anxiety and depression-related behaviors in the open-field and forced swim tests. Because serotonin (5-HT) 5-HT 1A receptors seem to play an important role in the pathophysiology of major depression and in the mechanism of action of antidepressants, we investigated if 5-HT 1A receptor function is altered in deprived rats. Although the hypothermic response to the 5-HT 1A receptor agonist 8-OH-DPAT was increased in adult deprived rats compared to non-deprived control group, no differences between groups were found in the effect of the systemic 8-OH-DPAT administration on serotoninergic cell firing in dorsal raphe nucleus and in the 5-HT release at the ventral hippocampus levels. These results suggest that 5-HT 1A receptors are not substantially affected in adult Sprague–Dawley rats that were subjected to a maternal deprivation 3 h daily during the neonatal period.

Myenteric denervation differentially reduces enteroendocrine serotonin cell population in rats during postnatal development

The enteric nervous and enteroendocrine systems regulate different processes in the small intestine. Ablation of myenteric plexus with benzalkonium chloride (BAC) stimulates epithelial cell proliferation, whereas endocrine serotonin cells may inhibit the process. To evaluate the connection between the systems and the influence of myenteric plexus on serotoninergic cells in rats during postnatal development, the ileal plexus was partially removed with BAC. Rats were treated at 13 or 21 days and sacrificed after 15 days. The cell bodies of myenteric neurons were stained by beta NADH-diaphorase to detect the extension of denervation. The number of enteroendocrine cells in the ileum was estimated in crypts and villi in paraffin sections immunostained for serotonin. The number of neurons was reduced by 27.6 and 45% in rats treated on the 13th and 21st days, respectively. We tried to establish a correlation of denervation and the serotonin population according to the age of treatment. We observed a reduction of immunolabelled cells in the crypts of rats treated at 13 days, whereas this effect was seen in the villi of rats denervated at 21 days. These results suggest that the enteric nervous system might control the enteroendocrine cell population and this complex mechanism could be correlated to changes in cell proliferation.

5-HT 1A receptor activation counteracts c-Fos immunoreactivity induced in serotonin neurons of the raphe nuclei after immobilization stress in the male rat

The serotoninergic system and the 5-HT 1A receptors have been involved in the brain response to acute stress. The aim of our study was evaluate the role of the 5-HT 1A receptors in serotoninergic cells of rostral and caudal raphe nuclei under acute immobilization in rats. Double immunocytochemical staining of 5-hydroxy-tryptamine and c-Fos protein and stereology techniques were used to study the specific cell activation in the raphe nuclei neurons in five groups (control group, immobilization group (immobilization lasting 1 h), DPAT group (8-OH-DPAT 0.3 mg/kg, s.c.), DPAT + IMMO group (8-OH-DPAT 0.3 mg/kg, s.c., 30′ prior acute immobilization) and WAY + DPAT + IMMO group (WAY-100635 0.3 mg/kg, s.c. and 8-OH-DPAT 0.3 mg/kg, s.c., 45′ and 30′, respectively, before immobilization). Our results showed an increase in the number of c-Fos immunoreactive nuclei in serotoninergic cells in both dorsal and median raphe nuclei in the immobilized group. The 8-OH-DPAT pretreatment counteracted the excitatory effects of the acute immobilization in these brain regions. In addition, WAY-100635 administration reduced the effect of 8-OH-DPAT injection, suggesting a selective 5-HT 1A receptor role. Raphe pallidus and raphe obscurus did not show any differences among experimental groups. We suggest that somatodendritic 5-HT 1A receptors in rostral raphe nuclei may play a crucial role in both mediating the consequences of uncontrollable stress and the possible beneficial effects of treatment with 5-HT 1A receptor agonists.

Identification of Wake-Active Dopaminergic Neurons in the Ventral Periaqueductal Gray Matter

Recent evidence suggests that dopamine plays an important role in arousal, but the location of the dopaminergic neurons that may regulate arousal remains unclear. It is sometimes assumed that the dopaminergic neurons in the ventral tegmental area that project to the prefrontal cortex and striatum may regulate the state of arousal; however, the firing of these dopaminergic neurons does not correlate with overall levels of behavioral wakefulness. We identified wake-active dopaminergic neurons by combining immunohistochemical staining for Fos and tyrosine hydroxylase (TH) in awake and sleeping rats. Approximately 50% of the TH-immunoreactive (TH-ir) cells in the ventral periaqueductal gray matter (vPAG) expressed Fos protein during natural wakefulness or wakefulness induced by environmental stimulation, but none expressed Fos during sleep. Fos immunoreactivity was not seen in the substantia nigra TH-immunoreactive cells in either condition. Injections of 6-hydroxydopamine into the vPAG, which killed 55-65% of wake-active TH-ir cells but did not injure nearby serotoninergic cells, increased total daily sleep by approximately 20%. By combining retrograde and anterograde tracing, we showed that these wake-active dopaminergic cells have extensive reciprocal connections with the sleep-wake regulatory system. The vPAG dopaminergic cells may provide the long-sought ascending dopaminergic waking influence. In addition, their close relationship with the dorsal raphe nucleus will require reassessment of previous studies of the role of the dorsal raphe nucleus in sleep, because many of those experiments may have been confounded by the then-unrecognized presence of intermingled wake-active dopaminergic neurons.

Darkness induced neuroplastic changes in the serotoninergic system of the chick retina

Sensory experience is critical for the formation of neuronal circuits and it is well known that neuronal activity plays a crucial role in the formation and maintenance of synapses. In the vertebrate retina, exposure to different environmental conditions results in structural, physiological, neurochemical and pharmacological changes. Serotoninergic (5HT) amacrine cells of the chicken retina are bistratified interneurons whose primary dendrites descend through the inner nuclear layer (INL) to branch in the inner plexiform layer (IPL) forming two plexi, an outer network, localized to sublamina 1, and an inner network, localized to sublamina 4 and 5 of the IPL. Their development is temporally correlated with the establishment of synapses in the retina and with the emergence of the typical adult electroretinogram. It is unknown, however, which role these cells play in processing visual information and whether visual deprivation modifies their phenotype. Here, we show that, in the chicken, red-light rearing from hatching to postnatal day 12 significantly alters the stratification pattern of 5HT amacrine cells, inhibiting their age-dependent pruning measured with morphometric and densitometric procedures; as well as increasing serotonin immunoreactivity measured as relative optical density. This change in dendritic arborization, accompanied by an increase in serotonin concentration in dark adapted conditions, may decrease visual threshold, thus increasing visual sensitivity.

3,4-metilenodioximetanfetamina (‘éxtasis’): efectos emocionales y cognitivos a largo plazo y depleción serotoninérgica

The main objective of this study is to describe the different neuropsychological deficits associated to the consumption of 3,4-methylenedioxymethamphetamine (MDMA or 'ecstasy'), as well as the growing evidence that attributes these deficits to the selective axonal damage to serotoninergic cells brought about by this substance.MDMA is an amphetamine derivative that, like its precursor, has properties as a stimulant. Part of its chemical structure is similar to that of the hallucinogen mescaline with which it shares the capacity to alter perception. Nevertheless, the primary pharmacological effect of this substance, which is what usually leads to its use and abuse, is chiefly linked to an intense positive emotional state. This effect on the individual's mood is also usually accompanied by numerous feelings of empathy, sociability and closeness, which turn this drug into a powerful entactogenic agent (a term used in psychotherapy to describe a state of wellbeing, closeness and emotional self-awareness produced by certain compounds). The antidepressant and entactogenic effects induced by an acute dose of MDMA can be accounted for by the notable increase in serotonin bioavailability triggered by the drug. Repeated consumption of MDMA, however, ends up affecting many functions that have been related to the serotoninergic systems, such as sleep, appetite, attention and memory, or one's emotional state.Most of the neuropsychological disorders found in individuals who take ecstasy on a regular basis can be explained by the selective neurodegeneration processes that the drug appears to produce in the serotonin terminals of the brain in the long run.

Circadian change in tryptophan hydroxylase protein levels within the rat intergeniculate leaflets and raphe nuclei

Serotonin (5-HT) is involved in the synchronisation of the mammalian circadian clock located in the suprachiasmatic nuclei of the hypothalamus (SCN). This clock is synchronised by light (photic cues) and by non-photic cues. Non-photic cues are notably conveyed to the SCN by a direct 5-HT pathway arising from the mesencephalic median raphe nucleus (MRN). Furthermore, an indirect projection conveys non-photic inputs by 5-HT fibres from the mesencephalic dorsal raphe nucleus (DRN) to the intergeniculate leaflets of the thalamus (IGL) which project to the SCN. In the rat, the quantitative distribution of tryptophan hydroxylase (TpH), used as an index of 5-HT synthesis, was studied by in situ immunoautoradiography in both the serotoninergic cell bodies area of the raphe nuclei and the serotoninergic terminal field of the IGL. Under a 12 h light: 12 h dark (LD 12:12), TpH protein amount exhibited a rhythmic variation within the IGL. The maximum levels were reached at the day/night transition. In both MRN and the lateral groups of the DRN, TpH variations were opposite to those observed in the IGL. Such phase opposition was reported previously in the MRN/SCN pathway and was correlated with a rhythmic release of 5-HT within the SCN [Eur J Neurosci 15 (2002) 833]. Thus, the daily rhythmicity of TpH levels observed in DRN-IGL pathway may be correlated with a rhythmic release of 5-HT in the IGL at the beginning of the night. Under constant darkness, TpH rhythmic variations in the two serotoninergic pathways were maintained and similar to those observed under light/dark cycle. These results demonstrate the existence of a circadian endogenous functioning in the 5-HT neurones projecting to the rat circadian system.