Neuromodulatory role of serotonin in the anticonvulsant activity of 2-phenylbenzoate of 3-diethylamino-1-propanol.HCl (JAW-669)

The role of serotonin in the mediation of the anticonvulsant activity of JAW-669 was investigated against maximal electric shock (MES)-induced seizures in mice. A dose-dependent protection against seizures was provided by JAW-669 (4, 6 and 8 mg/kg, IP) and the calculated ED50 value was 6.01 mg/kg, IP. Pretreatment of mice with 5-hydroxytryptophan (50 mg/kg, IP) 2 hr before the administration of JAW-669 (6.01 mg/kg, IP) was found to cause a 40% increase in the ability of JAW-669 to provide protection against MES-induced seizures. Similar pretreatment with tryptophan (100 mg/kg, IP, 1 hr) caused a 30% decrease in the anticonvulsant activity of JAW-669. Prior administration of p-chlorophenylalanine (300 mg/kg, IP, 48 hr) and methysergide (10 mg/kg, IP; 0.5 hr) before administration of JAW-699 caused a 66% and 74% decrease, respectively, in the ability of JAW-669 to provide protection against MES-induced seizures. These results suggest a facilitatory role of serotonin in the anticonvulsant activity of JAW-669.     

Calcium-independent afterdepolarization regulated by serotonin in anterior thalamus

Previous studies have identified an afterdepolarization (ADP) in thalamocortical neurons that is mediated by an upregulation of the hyperpolarization-activated current I(h). This ADP has been suggested to play a key role in the generation of spindle oscillations. In the lateral geniculate nucleus, upregulation of I(h) has been shown to be signaled by a rise in intracellular calcium leading to the activation of adenylate cyclase and formation of cAMP. However, it is unclear how generalizable this mechanism is to other thalamic nuclei. We have used whole cell recording to examine the electrophysiological properties of neurons of the anterodorsal thalamic nucleus, a nucleus thought not to undergo spindle oscillations. We now report that cells in this nucleus also display an ADP mediated by I(h). Surprisingly, the ADP and the underlying upregulation of I(h) persisted even after buffering intracellular calcium and blocking calcium influx. These results indicate that, in neurons of the anterodorsal thalamic nucleus, an I(h)-mediated ADP can occur through a mechanism that does not involve a rise in intracellular calcium. We next examined the possibility that this calcium-independent ADP might be modulated by serotonin. Serotonin produced a robust enhancement in the amplitude of the ADP even after strong buffering of intracellular calcium and blockade of calcium channels. These results indicate that neurons of the anterodorsal thalamic nucleus display a calcium-independent, I(h)-mediated ADP and that this ADP is a target for regulation by serotonin. These findings identify a novel mechanism by which serotonin can regulate neuronal excitability.     

Neuromodulatory transmitter systems in the cortex and their role in cortical plasticity

Cortical neuromodulatory transmitter systems refer to those classical neurotransmitters such as acetylcholine and monoamines, which share a number of common features. For instance, their centers are located in subcortical regions and send long projection axons to innervate the cortex. The same transmitter can either excite or inhibit cortical neurons depending on the composition of postsynaptic transmitter receptor subtypes. The overall functions of these transmitters are believed to serve as chemical bases of arousal, attention and motivation. The anatomy and physiology of neuromodulatory transmitter systems and their innervations in the cerebral cortex have been well characterized. In addition, ample evidence is available indicating that neuromodulatory transmitters also play roles in development and plasticity of the cortex. In this article, the anatomical organization and physiological function of each of the following neuromodulatory transmitters, acetylcholine, noradrenaline, serotonin, dopamine, and histamine, in the cortex will be described. The involvement of these transmitters in cortical plasticity will then be discussed. Available data suggest that neuromodulatory transmitters can modulate the excitability of cortical neurons, enhance the signal-to-noise ratio of cortical responses, and modify the threshold for activity-dependent synaptic modifications. Synaptic transmissions of these neuromodulatory transmitters are mediated via numerous subtype receptors, which are linked to multiple signal transduction mechanisms. Among the neuromodulatory transmitter receptor subtypes, cholinergic M(1), noradrenergic beta(1) and serotonergic 5-HT(2C) receptors appear to be more important than other receptor subtypes for cortical plasticity. In general, the contribution of neuromodulatory transmitter systems to cortical plasticity may be made through a facilitation of NMDA receptor-gated processes.     

Temporal properties of feedforward and feedback pathways between the thalamus and visual cortex in the ferret

This study examines the temporal properties of geniculocortical and corticogeniculate (CG) pathways that link the lateral geniculate nucleus (LGN) and primary visual cortex in the ferret. Using electrical stimulation in the LGN to evoke action potentials in geniculocortical and CG axons, results show that conduction latencies are significantly faster in geniculocortical neurons than in CG neurons. Within each pathway, axonal latency and visual physiology support the view of sub-classes of neurons. By examining the timing of visual responses and the latency of CG feedback, estimates indicate that visual information can reach the cortex and return to the LGN as early as 60 msec following the onset of a visual stimulus. These findings place constraints on the functional role of corticogeniculate feedback for visual processing.     

Neuromodulatory role of acetylcholine in visually-induced cortical activation: behavioral and neuroanatomical correlates

Acetylcholine is released in the primary visual cortex during visual stimulation and may have a neuromodulatory role in visual processing. The present study uses both behavioral and functional neuroanatomy investigations to examine this role in the rat. In the first set of experiments the cholinergic system was lesioned with 192 immunoglobulin G (IgG) saporin and the visual acuity and performance in a visual water maze task were assessed. The cholinergic lesion did not affect the visual acuity measured pre- and post-lesion but it did reduce the efficiency to learn a novel orientation discrimination task measured post-lesion. In order to better understand the involvement of the cholinergic system in the neuronal activity in the visual cortex c-Fos expression induced by patterned visual stimulation was further investigated. Results obtained following lesion of the cholinergic fibers (192 IgG-saporin or quisqualic acid), muscarinic inhibition (scopolamine), or NMDA receptor inhibition (CPP) were compared with control conditions. Double and triple immunolabeling was used in order to determine the neurochemical nature of the activated cortical cells. The results demonstrated that patterned stimulation elicited a significant increase in c-Fos immunolabeled neurons in layer IV of the contralateral primary visual cortex to the stimulated eye which was completely abolished by cholinergic fibers lesion as well as scopolamine administration. This effect was independent of NMDA receptor transmission. The c-Fos activation was predominantly observed in the glutamatergic spiny stellate cells and less frequently in GABAergic interneurons. Altogether, these results demonstrate a strong involvement of the basal forebrain cholinergic system in the modulation of post-synaptic visual processing, which could be related to cognitive enhancement or attention during visual learning.     

Evidence for a projection from the perireticular thalamic nucleus to the dorsal thalamus in the adult rat and ferret

Summary During early development, the perireticular thalamic nucleus is very large (i.e. has many cells) and has a strong projection to the dorsal thalamus and to the cerebral neocortex. By adulthood, the nucleus has much reduced in size and only a few cells remain. It is not clear whether these perireticular cells that remain into adulthood maintain their connections with the dorsal thalamus and with the neocortex. This study examines this issue by injecting neuronal tracers into various nuclei of the dorsal thalamus (dorsal lateral geniculate nucleus, medial geniculate complex, ventroposteromedial nucleus, lateral posterior nucleus, posterior thalamic nucleus) and into different areas of the neocortex (somatosensory, visual, auditory). After injections of tracer into the individual nuclei of the rat and ferret dorsal thalamus, retrogradely-labelled perireticular cells are seen. In general, after each injection, the retrogradely-labelled perireticular cells lie immediately adjacent to a group of retrogradely-labelled reticular cells. For instance, after injections into the medial geniculate complex, perireticular cells adjacent to the auditory reticular sector are retrogradely-labelled, whilst after an injection into the dorsal lateral geniculate nucleus, retrogradely-labelled perireticular cells adjacent to the visual reticular sector are seen. By contrast, injections of tracer into various areas of the rat and ferret neocortex result in no retrogradely-labelled cells in the perireticular nucleus. Thus, unlike during perinatal development when perireticular cells project to both neocortex and dorsal thalamus, perireticular cells in the adult seem to project to the dorsal thalamus only: the perireticular projection to the neocortex appears to be entirely transient.     

Connecting thalamus and cortex: the role of ephrins

The complex task of wiring up the brain during embryonic development is achieved by a multitude of guidance signals acting in complex combinations to drive growing axons to their proper targets. The somatosensory system provides an extensively studied model system featuring many universal mechanisms of neural development. In rodents, it constitutes an important model to study how precise topographic connections are achieved. Recent evidence suggests that the Eph/ephrin family of guidance molecules is of pivotal importance for the development of the somatosensory system. Members of Eph/ephrin family are thought to be involved in the global presorting of thalamic axons projecting to the cortex, in labeling specific cortical areas for innervation, in providing topographic cues within the target area, and in distinguishing cortical layers for intracortical wiring. The Eph/ephrin system also seems to contribute to the formation of specific corticothalamic feedback projections. So far, the functions of only a few members of the Eph/ephrin family have been examined, but expression analysis indicates complex combinatorial effects of these signaling molecules. Understanding the Eph/ephrin wiring code is expected to yield new insights into the development and plasticity of brain circuits involved in higher functions.     

Neuromodulatory function of neuropeptides in the normal CNS

Neuropeptides are small protein molecules produced and released by discrete cell populations of the central and peripheral nervous systems through the regulated secretory pathway and acting on neural substrates. Inside the nerve cells, neuropeptides are selectively stored within large granular vesicles (LGVs), and commonly coexist in neurons with low-molecular-weight neurotransmitters (acetylcholine, amino acids, and catecholamines). Storage in LGVs is responsible for a relatively slow response to secretion that requires enhanced or repeated stimulation. Coexistence (i.e. the concurrent presence of a neuropeptide with other messenger molecules in individual neurons), and co-storage (i.e. the localization of two or more neuropeptides within individual LGVs in neurons) give rise to a complicated series of pre- and post-synaptic functional interactions with low-molecular-weight neurotransmitters.The typically slow response and action of neuropeptides as compared to fast-neurotransmitters such as excitatory/inhibitory amino acids and catecholamines is also due to the type of receptors that trigger neuropeptide actions onto target cells. Almost all neuropeptides act on G-protein coupled receptors that, upon ligand binding, activate an intracellular cascade of molecular enzymatic events, eventually leading to cellular responses. The latter occur in a time span (seconds or more) considerably longer (milliseconds) than that of low-molecular-weight fast-neurotransmitters, directly operating through ion channel receptors. As reviewed here, combined immunocytochemical visualization of neuropeptides and their receptors at the ultrastructural level and electrophysiological studies, have been fundamental to better unravel the role of neuropeptides in neuron-to-neuron communication.     

Relaxation in ferret ventricular myocytes: role of the sarcolemmal Ca ATPase

In ferret ventricular myocytes the rate of intracellular Ca concentration [Ca]_i decline and relaxation is remarkably fast (compared with rabbit and rat) under conditions where both the sarcoplasmic reticulum Ca uptake and Na/Ca exchange are inhibited. Here we explore the possibility that this rapid [Ca]_i decline in ferret cells is attributable to the sarcolemmal Ca ATPase by using carboxyeosin (a potent inhibitor of the sarcolemmal Ca-ATPase). We compare the effects of carboxyeosin with those of elevated extracellular [Ca] ([Ca]_o) (a thermodynamic approach to limit Ca transport by the sarcolemmal Ca ATPase). In rabbit cells, carboxyeosin and high [Ca]_o slowed [Ca]_i decline similarly and both virtually abolished [Ca]_i decline when mitochondrial Ca uptake was also inhibited. In ferret cells, carboxyeosin treatment produced these same effects on [Ca]_i decline, but high [Ca]_o did not mimic them. Moreover, only in carboxyeosintreated ferret cells did additional inhibition of mitochondrial Ca uptake nearly abolish [Ca]_i decline. We conclude that, carboxyeosin loading can inhibit the sarcolemmal Ca-ATPase in intact myocytes; that this pump seems likely to be responsible for the much faster relaxation observed in ferret cells after block of SR Ca accumulation and Na/Ca exchange transport and that the sarcolemmal Ca pump apparently has different characteristics in rabbit and ferret ventricular myocytes. Present address: Centro de Engenharia Biomédica Caixa Postal 6040, Universidade Estadual de Campinas (UNICAMP), 13081 Campinas, SP, Brazil     

Studies on the neuroendocrine role of serotonin

The aim of the thesis was to investigate in male Wistar rats, the involvement of serotonin (5-HT) and 5-HT receptors in the regulation of the gene expression of hypothalamic hormones and in the secretion of the pituitary gland hormones prolactin (PRL), adrenocorticotropic hormone (ACTH), vasopressin (AVP) and oxytocin in basal and stress conditions. Furthermore, to study the significance of some distinctive central nuclei in these processes, and the metabolism of 5-HT in the hypothalamus and the dorsal raphe nucleus (DRN). The experiments were focused on (1) determination of involved neurons and nuclei (2) the hypothalamic level and (3) the pituitary gland level of regulation. The studies were typically performed in vivo but some studies were performed in vitro. Stereotactically neurotoxic lesion with 5,7-dihydroxy-5-HT in the dorsal raphe nucleus (DRN) or the hypothalamic paraventricular nucleus (PVN) reduced the ACTH and AVP response to stress, indicating an importance of these structures for this response. In situ hybridization on rat brain slices with oligopeptides showed an increase of corticotropin releasing hormone (CRH) mRNA in the PVN and proopiomelanocortin in the anterior pituitary lobe upon stimulation of the 5-HT1A, 5-HT1B, 5-HT2A and 5-HT2C receptors. Stimulation of 5-HT2A+2C receptors increased AVP mRNA in the PVN but not in the supraoptic nucleus (SON), whereas the level of oxytocin (OT) mRNA was increased both in the SON and the PVN and this effect was in addition mediated via 5-HT1A+1B receptors. Serotonin infused directly into the PVN by microdialysis stimulated local release of AVP. CRH was found to have a major role but not a complete responsibility in the 5-HT-induced release of ACTH, since immunoneutralisation of CRH inhibited the POMC gene expression and the ACTH response and since 5-HT and 5-HT antagonists were able to modulate the ACTH release from anterior pituitary gland cells in vitro. Through the years of investigation, the classification of the 7 main groups of 5-HT receptors (5-HT1 - 5-HT7) has changed due to molecular biological characterisation of the receptors and new receptors have been identified. With a battery of 5-HT agonists and antagonists several pharmacological experiments were performed with systemically or central administration of compounds and radioimmuno assay of plasma for pituitary gland hormone levels. Specific substances were not available for all 5-HT receptors and subreceptors thus some conclusions are a based on combination of experiments. The 5-HT induced PRL response is mediated via 5-HT1A, 5-HT2A, 5-HT2C and 5-HT3 receptors. In addition an involvement of 5-HT1B, 5-HT5 or 5-HT7 receptors seem possible. The ACTH response to 5-HT is mediated via 5-HT1A, 5-HT1B, 5-HT2A and 5-HT2C receptors and an involvement of the 5-HT4, 5-HT5 and 5-HT7 receptors is proposed. Peripheral secretion of AVP upon stimulation with 5-HT is mediated via 5-HT2C, 5-HT4 and 5-HT7 receptors but not 5-HT1A receptors. The secretion of OT is primarily mediated via 5-HT1A, 5-HT2C and 5-HT4 receptors and probably also 5-HT1B, 5-HT2A, 5-HT5A and 5-HT7 receptors. Physical and psychological stress activates hippocampal and hypothalamic 5-HT neurons. In contrast to other stress factors, restraint stress increases the content of 5-HT in the DRN but do not increase the metabolism of 5-HT and does not induce changes in hypothalamic levels of 5-HT. Large variations are found in the literature with different kinds of stress, different measurements and different time schedules. Restraint or ether stress induced secretion of PRL involves 5-HT2 and 5-HT3 receptors, whereas the ACTH secretion is mediated via 5-HT1A, 5-HT2A and 5-HT2C receptors. In the present study restraint stress increased AVP secretion, but opposite findings has reported possibly due to differences in the stress procedure. The 5-HT2, 5-HT3 and 5-HT4 receptor is involved in the AVP response to restraint whereas the OT response involves the 5-HT1A and the 5-HT2 receptor. The 5-HT2 receptor is involved in the OT response to dehydration or haemorrhage, whereas the AVP responses to these stressors probably do not involve 5-HT. It can be concluded that 5-HT is involved in basal and stress-induced regulation of PRL, ACTH, AVP and oxytocin mainly via the 5-HT2A+2C receptors but other receptors are also important but differs from hormone to hormone. Serotonin affect the secretion of CRH and ACTH both at the hypothalamic, pituitary portal and pituitary gland level, and possibly also at the adrenal level.     

The role of serotonin in hot flushes

Hot flushes are experienced in those periods of the female life when estrogen levels are low. Hormone replacement therapy is thus the first choice for treatment of hot flushes. However this treatment is not always accepted or contraindicated for a variety of reasons. Estrogen (and progestogen) strongly interact with a number of neurotransmitters and this has led to a range of non-hormonal treatments including compounds that act via the noradrenergic or dopaminergic systems as well as herbal remedies. These treatments (which are shortly reviewed) are not always successful. Surprisingly, apart from treatment with some selective serotonin (5-HT) reuptake inhibitors (SSRI's), up till now, little attention is given to the strong interaction of estrogens with the serotonergic system. These interactions are shortly reviewed. Based on these interactions, a hypothesis on the genesis of hot flushes is postulated. Especially the 5-HT_2A receptor subtype may play a key role in the occurrence of hot flushes. A number of arguments that support this hypothesis are discussed.     

A limited role for mediodorsal thalamus in devaluation tasks

Six experiments were performed to determine the role of mediodorsal thalamus (MD) in the devaluation task, varying the type of contingencies (Pavlovian or operant), the number of reinforcers (one vs. two), and the order of experiments (in na?ve or experimentally experienced rats). MD-lesioned rats were impaired in devaluation performance when switched between Pavlovian and operant devaluation tasks, but not when switched from one Pavlovian devaluation task to another Pavlovian devaluation task. MD lesions caused no devaluation impairment in a multiple-reinforcer Pavlovian devaluation task. These results suggest that MD lesions impair performance in devaluation tasks as a result of an inability to switch the form of associations made from one type of outcome-encoding association to another. This is in accord with previous literature suggesting that MD is needed for strategy set shifting. The results further suggest that MD is a necessary part of devaluation circuits only in cases in which previous associations need to be suppressed in order for new associations to be learned and control behavior, and otherwise the devaluation circuit does not require MD.     

Modulatory effects of serotonin on excitatory amino acid responses and sensory synaptic transmission in the ventrobasal thalamus

Excitatory amino acid receptors are thought to mediate sensory input to the ventrobasal thalamus. There is evidence for a brainstem serotonergic projection to the ventrobasal thalamus which may have a modulatory role. The possibility that serotonin may selectively modulate responses to excitatory amino acid receptor agonists, and its effects on sensory synaptic transmission has been examined in the rat ventrobasal thalamus in vivo. Iontophoretic ejection of serotonin at low currents produced a marked facilitation of responses to excitatory amino acids. In contrast, excitatory responses to cholinomimetic agonists were attenuated. Synaptic transmission was concomitantly enhanced or unchanged in these circumstances. Higher serotonin ejection currents reversed the facilitation, or inhibited excitatory amino acid responses and synaptic transmission. It is concluded that serotonin can modulate responses to excitatory amino acids relatively selectively and that synaptic transmission of somatosensory information through the ventrobasal thalamus may be susceptible to brainstem serotonergic modulation.     

Psychotogenic drugs and delirium pathogenesis: the central role of the thalamus

Delirium is thought to be a temporary psychiatric disorder resulting from a reduced central cholinergic transmission, combined with an increased dopaminergic transmission. The cholinergic and the dopaminergic systems interact not only with each other but with glutamatergic and gamma-amino-butyric acid (GABA) pathways. Besides the cerebral cortex, critical anatomical substrates of psychosis pathophysiology would comprise the striatum, the substantia nigra/ventral tegmental area, and the thalamus. The thalamus acts as a filter, allowing only the relevant information to travel to the cortex. Drugs of abuse (e.g. PCP, Ecstasy), as well as psychoactive medications frequently prescribed to hospitalized patients (e.g. benzodiazepines, opioids) could compromise the thalamic gating function, leading to sensory overload and hyperarousal. We propose that drug-induced delirium would result from the transient thalamic dysfunction caused by exposure to medications that interfere with central glutamatergic, GABAergic, dopaminergic and cholinergic pathways at critical sites of action. This model provides directions for future studies in neurophysiology, in vivo brain imaging, and psychopharmacology investigating delirium neuropathophysiology.     

The role of melatonin and serotonin in aging

The hypothesis presented in this paper defines aging as a pathological process originating in the pineal gland. This results in a diminished output of melatonin, along with a diminished melatonin to serotonin ratio, leading to a decline in adaptive processes and a predictable syndrome manifested by the "diseases of the aged" (DOA) and subsequent death of the organism. That is, aging is a syndrome of relative melatonin deficiency resulting from the gradual failure of the pineal gland.