Effect of thalamic parafascicularis nucleus stimulation in regulation of serotoninergic transmission in the cat caudate nucleus: involvement of autoreceptors in the dorsalis raphe nucleus

The mechanisms involved in parafascicularis nucleus control on serotoninergic neurons projecting into the caudate nucleus were investigated in "encéphale-isole" cats. The effects of unilateral stimulation of the parafascicularis nucleus on the release of newly synthesized [3H]serotonin were simultaneously determined in the ipsilateral caudate nucleus and the dorsalis raphe nucleus using push-pull cannulae. The actions of various pharmacological treatments performed either in the caudate nucleus or in the dorsalis raphe nucleus were also examined. The electrical or chemical stimulation of the parafascicularis nucleus induced a decrease in striatal [3H]serotonin release and an increase in [3H]serotonin release in the dorsalis raphe nucleus. The blockade of cholinergic (mecamylamine) and glutamatergic (PK 26124) transmissions at the striatal level did not modify the thalamic stimulation-induced effect on serotonin release in the caudate nucleus or in the dorsalis raphe nucleus. However, a decrease induced by parafascicularis nucleus stimulation in serotonin release in the caudate nucleus could not be observed when the autoreceptors present on serotoninergic nerve cell bodies localized in the dorsalis raphe nucleus were blocked by a methiothepin perfusion within the nucleus. These results indicate that the parafascicularis nucleus controls striatal serotonin transmission by inducing changes in the nerve activity of serotoninergic neurons in the dorsalis raphe nucleus via somatodendritic serotonin release and autoreceptors.     

Synaptic organization of the nucleus raphe dorsalis of the cat

Summary This electron microscopic study describes the different types of synaptic terminals found in the nucleus raphe dorsalis of the adult cat. Serial section analysis was used extensively to confirm the nature of the synaptic contact established by the various classes of terminals.Five different classes of terminals are identified according to the shape and packing density of the synaptic vesicles and type of contact they establish. The most common class (RDI-type) contains densely packed, round, agranular synaptic vesicles and establishes asymmetrical synaptic contacts. A second class (RDII-type) also contains spherical synaptic vesicles, but establishes symmetrical synaptic contacts with dendrites of all sizes. Most of the terminals in these two classes contain a few dense-cored synaptic vesicles, but a small sub-group contains many dense-cored vesicles. A third, less frequent, class (RSI-type) contains sparsely packed spherical synaptic vesicles and the majority of these terminals have asymmetrical contacts. A fourth terminal class contains pleomorphic synaptic vesicles (P-type), contacts dendrites of all sizes, and usually establishes symmetrical synaptic contacts. Finally, boutons thought to be the vesicle-filled excrescences of dendrites (postsynaptic dendrites) are found and in some cases the dendritic origin of these profiles was confirmed by serial sectioning. Such boutons containing pleomorphic vesicles are presynaptic to other such dendrites as well as conventional dendrites, and are postsynapticto the other terminal types described.Somata within the nucleus exhibit somatic spines but receive few synaptic contacts. Most axo-somatic terminals contain either round or pleomorphic vesicles and have postsynaptic thickenings intermediate to the symmetric and asymmetric types.     

Cooling of the nucleus raphe dorsalis induces sleep in the cat

Localized moderate cooling (+10°C) of the anterior and ventral part of the raphe dorsalis induced cortical synchronization and ponto-geniculo-occipital (PGO) waves followed sometimes by paradoxical sleep. Cooling the region of the raphe magnus produced cortical arousal. After depletion of brain 5-hydroxytryptamine (5-HT) by para-chlorophenylalanine, cooling the raphe dorsalis resulted in muscle atonia and bursts of high frequency PGO waves but not in cortical synchronization. Following restoration of brain 5-HT by intravenous administration of 5-hydroxytryptophan, cooling again produced cortical synchronization and PGO waves as in the control experiments, but with a diminishing efficacy during repetitive cooling episodes.     

In vivo evidence for acetylcholine control of serotonin release in the cat caudate nucleus: influence of halothane anaesthesia

Using a push-pull cannula technique and an isotopic method for the estimation of [3H]serotonin continuously synthesized from [3H]tryptophan, the effects of acetylcholine were investigated on the in vivo release of [3H]serotonin in the cat basal ganglia and the dorsal raphe nucleus. The unilateral striatal application of acetylcholine (5 x 10(-5) M) reduced local release of [3H]serotonin. This effect was mimicked by nicotine (5 x 10(-5) M) and prevented by mecamylamine (10(-6) M. Oxotremorine (5 x 10(-5) M) had no effect on the local release of [3H]serotonin. All these treatments failed to modify [3H]serotonin release in the ipsilateral substantia nigra or in the dorsal raphe nucleus. The superfusion of serotonergic nerve terminals of the caudate nucleus with tetrodotoxin prevented the inhibitory acetylcholine-induced effect on serotonin release. Furthermore, bicuculline (5 x 10(-5) M) in the caudate nucleus blocked the effect of nicotine, while gamma-aminobutyric acid (10(-5) M) induced a decrease in local release of [3H]serotonin. These data strongly suggest that the inhibitory control exerted by acetylcholine on serotonergic transmission could involve gamma-aminobutyric acid interneurons. Acetylcholine-induced changes in [3H]serotonin release were only observed in non-anaesthetized "encéphale isolé" cats and not in halothane-anaesthetized animals. The possibility that such a regulation could be presynaptic (direct or through other neurotransmitters) or related to a change in the activity of the serotonergic raphe-striatal neuronal system is discussed.     

Regulation of dorsal raphe nucleus function by serotonin autoreceptors: a behavioral perspective

Neurotransmission by serotonin (5-HT) is tightly regulated by several autoreceptors that fine-tune serotonergic neurotransmission through negative feedback inhibition at the cell bodies (predominantly 5-HT(1A)) or at the axon terminals (predominantly 5-HT(1B)); however, more subtle roles for 5-HT(1D) and 5-HT(2B) autoreceptors have also been detected. This review provides an overview of 5-HT autoreceptors, focusing on their contribution in animal behavioral models of stress and emotion. Experiments targeting 5-HT autoreceptors in awake, behaving animals have generally shown that increasing autoreceptor feedback is anxiolytic and rewarding, while enhanced 5-HT function is aversive and anxiogenic; however, the role of serotonergic activity in behavioral models of helplessness is more complex. The prevailing model suggests that 5-HT autoreceptors become desensitized in response to stress exposure and antidepressant administration, two seemingly opposite manipulations. Thus there are still unresolved questions regarding the role of these receptors-and serotonin in general-in normal and pathological states.     

Enkephalin-immunoreactive perikarya in the cat raphe dorsalis

The serotonin-containing nucleus raphe dorsalis (RD) of the cat contains numerous leucine-enkephalin immunoreactive cells, throughout its rostral-caudal extent. The distribution of the enkephalin neurons closely parallels the cytoarchitectural boundaries of the RD, as described in previous Nissl preparations. Enkephalin perikarya are most numerous along the midline of the RD, but also extend ventrally, into the dorsal portion of the nucleus centralis superior, and laterally, into the ‘wings’ of the rostral RD, at the level of the IV nucleus. The possible contribution of these enkephalin cells to endogenous pain control systems is discussed.     

Regularity of the spontaneous discharge of neurons in the nucleus raphe dorsalis of the cat

Impulse discharges were recorded from 43 neurons in the nucleus raphe dorsalis of the cat. Of these neurons examined for spontaneous activity, 23 fired in remarkably regular rhythm (clock-like neurons); 18 fired irregularly (non-clock-like neurons); and 2 fired with an unclassified pattern. Almost all clock-like neurons were unresponsive to any kind of peripheral somatic stimuli, while the majority of non-clock-like neurons were excited by both nociceptive and non-nociceptive stimuli or by non-nociceptive stimuli. The somatic receptive fields were large, spanning the entire body.     

Sleep in the cat: raphe dorsalis and vasotocin

To determine the relationship between the raphe nuclei and the sleep-inducing pineal peptide arginine vasotocin (AVT), we investigated the hypnogenic effects of intraventricularly administered AVT in adult cats with electrolytic lesions performed either in the raphe dorsalis nucleus (RDN) or the raphe pontis nucleus (RPN). The following results were observed: (a) RDN lesions produced a quantitative decrease in NREM sleep and a transitory increase in REM sleep and narcoleptic-like alterations of REM sleep in the first 2-5 postoperative days. On the eighth postoperative day, normal (prelesion) sleep parameters were restored. (b) RDN lesions prevented AVT from inducing its hypnogenic effects during the first 5 postoperative days, but the AVT hypnogenic effects reappeared when sleep parameters reached prelesion levels. (c) Although the RPN lesions produced a slight but statistically significant REM sleep decrease, this did not prevent AVT from inducing its hypnogenic effects. We conclude that (a) the anatomic site of AVT action within the brain may be related to RDN; (b) at least some of the sleep disturbances observed after RDN lesions (narcolepsy) may be due to damage of the AVT site of action or to modifications of AVT synthesis or release; (c) RDN belongs to a large neural circuit that includes the pineal gland and habenula, a circuit that may play an important role in the sleep-wake cycle by "fine-tuning" sleep mechanisms. We hypothesize that AVT could be the specific inhibitory neuromodulator of this neural circuit.     

Vesicle-containing dendrites in the nucleus raphe dorsalis of the cat. A serial section electron microscopic analysis

Summary The nucleus raphe dorsalis of the cat was examined by serial section electron microscopy and the presence of vesicle-containing dendrites is reported. Such dendrites were divided into two classes according to their synaptic contact. Dendrites containing round and/or pleomorphic vesicles associated with a clear synaptic specialization which was generally intermediate between a Gray's type I or II were classified as presynaptic dendrites. These presynaptic dendrites were presynaptic to conventional dendrites and dendritic spines. In addition, some profiles containing a sparse population of vesicles which may be dendritic in nature were observed involved in serial synaptic arrangements. A second class of dendrites were characterized by the presence of vesicles which were never found associated with any synaptic membrane specialization. Commonly, the vesicles were densely packed and associated with unusual densities. Serial section analysis of these densities showed that they were not presynaptic dense projections. We suggest that the existence of vesicle-containing dendrites in the nucleus raphe dorsalis of the cat constitute the morphological support for the dendritic release of neurotransmitters.     

Has the raphe dorsalis nucleus an asymmetric function?

Summary Differential pulse voltammetry (DPV) was used to measure 5-hydroxyindoles in the striata of anaesthetized rats. The amplitude of the voltammetry peaks in both striata decreased when 5-hydroxytryptophan (5-HTP) or 5-hydroxytryptamine (5-HT) was injected into the left side of the raphe dorsalis nucleus (RDN), but increased when either substance was injected into the right side. These findings indicate that the 5-hydroxyindole based communication between the striata and the RDN has a lateral asymmetry. The data are discussed with reference to histological observations suggesting a higher density of 5-hydroxyindole containing cell bodies in the lateral region than in the midline of this unpaired nucleus.     

In vivo assessment of the midbrain raphe nuclear regulation of serotonin release in the hamster suprachiasmatic nucleus

Serotonin (5-HT) plays important regulatory roles in mammalian circadian timekeeping; however, little is known concerning the regulation of serotonergic activity in the circadian clock located in the suprachiasmatic nuclei (SCN). By using in vivo microdialysis to measure 5-HT release we demonstrated that electrical or pharmacological stimulations of the dorsal or median raphe nuclei (DRN and MRN, respectively) can alter basal release of 5-HT in the hamster SCN. There were similar increases in SCN 5-HT release after electrical stimulation of either the MRN or DRN, indicating that both could contribute to the serotonergic activity in the SCN. Systemic pretreatment with the 5-HT antagonist metergoline abolished DRN-induced SCN 5-HT release but had little effect on MRN-induced SCN 5-HT release, suggesting different pathways for these nuclei in regulating 5-HT output in the SCN. Microinjections of the 5-HT1A autoreceptor agonist 8-OH-DPAT or antagonist WAY 100635 into the MRN caused significant inhibition and stimulation of SCN 5-HT release, respectively. Both drugs had substantially less effect in the DRN. These differential drug actions indicate that somatodendritic 5-HT1A autoreceptors on MRN neurons provide the prominent raphe autoregulation of 5-HT output in the SCN. Collectively the current results are evidence that DRN as well as MRN neurons can contribute to the regulation of 5-HT release in the hamster SCN. On the basis of the current observations and those from recent anatomic tracing studies of serotonergic projections to SCN it is hypothesized that DRN input to the SCN could be mediated by a DRN --> MRN --> SCN pathway involving a 5-HT-sensitive multisynaptic interaction between the DRN and MRN neurons.     

Collateral projections of nucleus raphe dorsalis neurones to the caudate-putamen and region around the nucleus raphe magnus and nucleus reticularis gigantocellularis pars alpha in the rat

It was examined whether or not the nucleus raphe dorsalis (RD) neurons projecting to the caudate-putamen (CPu) might also project to the motor-controlling region around the nucleus raphe magnus (NRM) and nucleus reticularis gigantocellularis pars alpha (Gia) in the rat. Single RD neurons projecting to the CPu and NRM/Gia by way of axon collaterals were identified by the retrograde double-labeling method with fluorescent dyes, Fast Blue and Diamidino Yellow, which were injected respectively into the CPu and NRM/Gia. Then, serotonin (5-HT)-like immunoreactivity of the double-labeled RD neurons was examined immunohistochemically; approximately 60% of the double-labeled RD neurons showed 5-HT-like immunoreactivity. The results indicated that some of serotonergic and non-serotonergic RD neurons might control motor functions simultaneously at the levels of the CPu and NRM/Gia by way of axon collaterals.     

Hippocampal 5-HT synthesis and release in vivo is decreased by infusion of 8-OHDPAT into the nucleus raphe dorsalis

Infusion of the serotonin-1A (5-HT_1A) agonist, 8-hydroxy-2-di(n-propylamino)tetralin (8-OHDPAT) (0.5, 1.0 and 2.0 μg) into the dorsal raphe, decreased extracellular 5-HT as measured by dialysis in the ventral hippocampus and also decreased 5-HT synthesis in both the hippocampus and the rest of the brain as measured by 5-hydroxytryptophan (5-HTP) accumulation following decarboxylase inhibition by NSD 1015.     

Unchanged density of 5-HT(1A) autoreceptors on the plasma membrane of nucleus raphe dorsalis neurons in rats chronically treated with fluoxetine

5-HT(1A) autoreceptors regulate the firing of 5-HT neurons and their release of 5-HT. In previous immuno-electron microscopic studies, we have demonstrated an internalization of 5-HT(1A) autoreceptors in the nucleus raphe dorsalis (NRD) of rats, after the acute administration of a single dose of the specific agonist 8-hydroxy-2-(di-n-propylamine)tetralin (8-OH-DPAT) or of the selective 5-HT reuptake inhibitor, fluoxetine. Twenty-four hours after either treatment, the receptors were back in normal density on the plasma membrane of NRD neurons. Here, we examined the subcellular localization of these receptors and the in vivo binding of the 5-HT(1A) radioligand 4,2-(methoxyphenyl)-1-[2-(N-2-pyridinyl)-p-fluorobenzamido]ethylpiperazine labeled with [(18)F]fluorine ([(18)F]MPPF) after chronic fluoxetine treatment (10 mg/kg daily for 3 weeks, by minipump). Unexpectedly, after such a treatment, there were no more differences between treated and control rats in either the density of plasma membrane labeling of NRD dendrites, or in the in vivo binding of [(18)F]MPPF, as measured with beta-microprobes. This was in keeping with earlier reports of an unchanged density of 5-HT(1A) receptor binding sites after chronic fluoxetine treatment, but quite unexpected from the strong electrophysiological and biochemical evidence for a desensitization of 5-HT(1A) autoreceptors under such conditions. Indeed, when the fluoxetine-treated rats were challenged with a single dose of 8-OH-DPAT, there was no internalization of the 5-HT(1A) autoreceptors, at variance with the controls. Interestingly, several laboratories have reported an uncoupling of 5-HT(1A) autoreceptors from their G protein in the NRD of rats chronically treated with fluoxetine. Therefore, the best explanation for our results is that, after repeated internalization and retargeting, functional 5-HT(1A) autoreceptors are replaced by receptors uncoupled from their G protein on the plasma membrane of NRD 5-HT neurons. Thus, the regulatory function of these autoreceptors may depend on a dynamic balance among their production, activation, internalization and recycling to the plasma membrane in inactivated (desensitized) form.     

中缝背核在睡眠-觉醒中的作用

中缝背核(DRN)是调控睡眠-觉醒的重要核团,在脑内有着广泛的神经投射区域。DRN位于中脑导水管腹侧,主要由5-羟色胺(5-HT)、γ-氨基丁酸(GABA)、多巴胺(DA)和谷氨酸(Glu)能4类神经元构成。本文主要介绍了DRN的各类神经元在睡眠-觉醒中相关作用的最新研究进展。     

Release of adenosine in vivo from cat caudate nucleus

A push-pull perfusion technique was used to study the release of endogenously synthesized [3H]-adenosine from caudate nucleus in the anaesthetized cat. The spontaneous release of [3H]adenosine newly synthesized from [3H]adenine reached a steady state level 40 min after the beginning of superfusion and continued for 4 h. Potassium and veratridine increased the release of newly synthetized [3H]-adenosine. The action of veratridine was completely blocked by tetrodotoxin. We conclude that spontaneous and evoked release of adenosine occurs in the cat striatum and might potentially affect central nervous function.