Some factors in the regulation of central serotonergic synapses

For a number of years release, reuptake, vesicular storage, and degradation via monoamine oxidase were the exclusive foci in chemical studies of the regulation of central serotonergic synapses. This brief review describes some other factors, including the brain levels of tryptophan and uptake processes in the nerve ending relevant to the substrate, the activity and amount of tryptophan hydroxylase in cell bodies and nerve endings, and a potential regulatable parasynaptic inactivation process, N-methylation. In chronic and acute drug studies some of these factors appear to function to compensate for acute perturbations in central serotonergic synaptic processes.     

REGIONAL TRANSPORT OF TRYPTOPHAN IN RAT BRAIN

Abstract— Tryptophan uptake was studied in brain slices and synaptosomes prepared from regions known to vary in the numbers of serotoninergic cell bodies and nerve endings that they contain. The rate of tryptophan uptake was highest in hypothalamus for both types of preparation. Differences among the regions were much more pronounced in isolated nerve endings (synaptosomes). Loading with tryptophan did not affect the uptake into tissue slices. Tryptophan accumulation in hypothalamus synaptosomes was reduced after intraventricular injection of 5,7–dihydroxytryptamine whereas no change was observed in synaptosomes prepared from cerebellum under the same conditions; accumulation by synaptosomes prepared from the hypothalamic and hippocampal regions was reduced after raphe lesions.     

Intrasynaptosomal conversion of tyrosine to dopamine as an index of brain catecholamine biosynthetic capacity

—The potential for intrasynaptosomal conversion of tyrosine to dopamine was evaluated in the cell bodies (substantia nigra) and nerve terminals (caudate-putamen) of the nigral-striatal dopaminergic pathway. The conversion technique involves measurement of ¹⁴CO₂ evolved from carboxyl-labelled tyrosine in the absence of both exogenous pteridine cofactor and DOPA decarboxylase. Evaluation of apparent K_m values for tyrosine uptake and conversion and observed maximal velocities suggest that conversion is not limited by movement of substrate into the synaptosomes. The results, based on brain regional and subcellular distribution, are consistent with the localization of conversion in nerve endings and suggest a rapid and reliable measure for catecholamine biosynthetic capacity when structural integrity of the nerve ending is maintained.     

Biochemistry and ultrastructure of serotonergic nerve endings in the lobster: serotonin and octopamine are contained in different nerve endings

In this article we report that the distribution of serotonin in the lobster nervous system parallels the distribution of octopamine and that the same tissues that contain endogenous serotonin can synthesize it from tryptophan. Octopamine and serotonin are highly concentrated in a neurosecretory region of the second thoracic roots in association with a group of neurosecretory cells. The roots possess separate high-affinity uptake systems for both serotonin and tryptophan. Radioactive serotonin, accumulated in tissues during incubations with either tritiated serotonin or tritiated tryptophan, can be released, in a calcium-dependent manner, by depolarization with potassium. A detailed morphological examination of the second thoracic roots shows four distinct categories of nerve endings in the vicinity of the neurosecretory cells. Octopamine is synthesized in one of these types of endings and serotonin in another. The high-affinity uptake systems for serotonin and tryptophan are found only in association with the endings that make serotonin. These endings and all the biochemical parameters of serotonin metabolism in the roots are selectively destroyed by previous injection of animals with the neurotoxin 5,7-dihydroxytryptamine.     

Uptake and Incorporation of Docosahexaenoic Acid (DHA) into Neuronal Cell Body and Neurite/Nerve Growth Cone Lipids: Evidence of Compartmental DHA Metabolism in Nerve Growth Factor-Differentiated PC12 Cells

Docosahexaenoic acid (DHA) accumulates in nerve endings of the brain during development. It is released from the membrane during ischemia and electroconvulsive shock. DHA optimizes neurologic development, it is neuroprotective, and rat adrenopheochromocytoma (PC12) cells have decreased PLA₂ activity when DHA is present. To characterize DHA metabolism in PC12 cells, media were supplemented with [³H]DHA or [³H]glycerol. Fractions of nerve growth cone particles (NGC) and cell bodies were prepared and the metabolism of the radiolabeled substrates was determined by thin-layer chromatography. [³H]glycerol incorporation into phospholipids indicated de novo lipid synthesis. [³H]DHA uptake was more rapid in the cell bodies than in the NGC. [³H]DHA first esterified in neutral lipids and later in phospholipids (phosphatidylethanolamine). [³H]glycerol primarily labeled phosphatidylcholine. DHA uptake was compartmentalized between the cell body and the NGC. With metabolism similar to that seen in vivo, PC12 cells are an appropriate model to study DHA in neurons.     

Morphometric analysis of hypoxia-induced synaptic activity in intrapulmonary neuroepithelial bodies

Summary A morphometric analysis has demonstrated ultrastructural changes induced by hypoxia in the epithelial cells and the intracorpuscular nerve endings of the presumed chemoreceptive intrapulmonary neuroepithelial bodies (NEB) of neonatal rabbits.Acute hypoxia stimulates an exocytosis of epithelial dense-core vesicles (DCV) at the level of the morphologically afferent or sensory (type 1 a) intracorpuscular nerve endings of the NEB. Assuming the epithelial cells to be chemoreceptive, this phenomenon could represent a transduction of sensory stimuli.In the morphologically efferent or motor (type 2 and type 1 b) intracorpuscular nerve endings of the NEB, acute hypoxia causes a depletion of synaptic vesicles and an increase in the amount of membrane-bounded cisternae and multivesicular bodies, suggestive of an enhanced synaptic activity of these nerve endings. It is proposed that the chemoreceptor cells could thus in turn be modulated centrifugally by their efferent-like intracorpuscular nerve endings.It has been proposed in our earlier studies that the NEB probably are intrapulmonary chemoreceptors with local secretory activities, reacting to the composition of the inhaled air. By the release of serotonin and peptide substances they may produce a local vasoconstriction in hypoxically aerated lung areas, enabling an intrapulmonary regulation of the V/Q ratio. The present study provides evidence that, in addition to this local effect, NEB could generate centripetal nerve impulses via exocytosis of epithelial DCV at the afferent-like intracorpuscular nerve endings. At the same time they could be modulated by the CNS via their efferent-like intracorpuscular nerve endings.With respect to their innervation, numerous similarities appear to exist morphologically and functionally between the carotid body and the intrapulmonary NEB.     

Effects of combined treatment with mephedrone and methamphetamine or 3,4-methylenedioxymethamphetamine on serotonin nerve endings of the hippocampus

Aims

Mephedrone is a stimulant drug of abuse with close structural and mechanistic similarities to methamphetamine and 3,4-methylenedioxymethamphetamine (MDMA). Although mephedrone does not damage dopamine nerve endings it increases the neurotoxicity of amphetamine, methamphetamine and MDMA. The effects of mephedrone on serotonin (5HT) nerve endings are not fully understood, with some investigators reporting damage while others conclude it does not. Presently, we investigate if mephedrone given alone or with methamphetamine or MDMA damages 5HT nerve endings of the hippocampus.

Main methods

The status of 5HT nerve endings in the hippocampus of female C57BL mice was assessed through measures of 5HT by HPLC and by immunoblot analysis of serotonin transporter (SERT) and tryptophan hydroxylase 2 (TPH2), selective markers of 5HT nerve endings. Astrocytosis was assessed through measures of glial fibrillary acidic protein (GFAP) (immunoblotting) and microglial activation was determined by histochemical staining with Isolectin B4.

Key findings

Mephedrone alone did not cause persistent reductions in the levels of 5HT, SERT or TPH2. Methamphetamine and MDMA alone caused mild reductions in 5HT but did not change SERT and TPH2 levels. Combined treatment with mephedrone and methamphetamine or MDMA did not change the status of 5HT nerve endings to an extent that was different from either drug alone.

Significance

Mephedrone does not cause toxicity to 5HT nerve endings of the hippocampus. When co-administered with methamphetamine or MDMA, drugs that are often co-abused with mephedrone by humans, toxicity is not increased as is the case for dopamine nerve endings when these drugs are taken together.     

Histochemical localization of acetylcholinesterase in the cerebral ganglia of Fasciola hepatica, a parasitic flatworm

Acetylcholinesterase activity was found in the cell bodies and extracellularly in the neuropile of the cerebral ganglia of the adult trematode parasite, Fasciola hepatica. Within neuronal cell bodies of the cerebral ganglion, acetylcholinesterase reaction product was found in the endoplasmic reticulum, in the cisternae of the Golgi apparatus, and in secretory vesicles near the inner (releasing face) cisternae. Acetylcholinesterase reaction product was not seen intracellularly within any nerve processes. The reaction product was found around the somatic cell membranes and in the extracellular space between closely apposed nerve processes in the neuropile. Acetylcholinesterase reaction product was associated with synaptic endings that contained clear spheroidal synaptic vesicles, and the reaction product was localized at the site of synaptic contact between the zone of apposition of the pre- and postsynaptic terminals. This intracellular and extracellular distribution of the enzyme is consistent with its function as the degrading enzyme in cholinergic transmission.     

Immunohistochemical demonstration of calbindin-containing nerve endings in the rat esophagus

Immunoreactivity for calbindin was found in nerve endings with irregular laminar shapes in the rat esophagus. In the myenteric ganglia, laminar endings of a range of sizes formed a complex network and appeared to lie at the surface of the ganglion. The myenteric ganglia that contained nerve endings were most abundant in the upper portion of the eosphagus, their number decreasing orally to anally. Calbindin-immunoreactive nerve cell bodies were scattered throughout the esophagus. Laminar terminals were found in the connective tissue of the lamina propria immediately beneath the epithelium and in the muscularis mucosae. Occasional nerve branches formed a network of aborizing endings that surrounded part of the submucosal arterioles. Immunoreactive nerve endings in the mucosa and submucosa were present only in the upper part of the cervical esophagus. Unilateral vagotomy caused a remarkable decrease in the number of the myenteric ganglia containing the calbindin-immunoreactive laminar endings after 15 days or survival; in some of ganglia, the laminar structures disappeared and nerve endings showing weak immunoreactivity had an indistinct appearance, so that the outline of the ganglia became obscure. In operated rats at 24 days, the number of innervated ganglia was about half that in normal rats. However, there was no change in the morphology and the occurrence of the immunoreactive laminar structures in the mucosa and submucosa after denervation. The results show that many of the laminar endings that are immunoreactive for calbindin in the myenteric ganglia are derived from the vagus nerve. Thus, the calbindin-immunoreactive nerve endings with laminar expansions that are found in the rat eosphageal wall could be sensory receptors.     

Postnatal maturation of neuroepithelial bodies and carotid body innervation: A quantitative investigation in the rabbit

The intrapulmonary airways contain oxygen-sensitive chemoreceptors which may be analogous to the arterial chemoreceptors: the neuroepithelial bodies (NEB). While the NEB are prominent in the neonatal lung, physiological studies indicate that the carotid bodies are still relatively inactive at birth. This points to an unequal degree of development of both during the early neonatal period. As a reflexogenic chemoreceptor function depends on a well-developed innervation, we undertook a comparative investigation of the development of the NEB and the carotid body glomus cell innervation. Two morphological aspects of the innervation of NEB and carotid body glomus cells were quantified in rabbits of different age groups. The total sectional area of intracorpuscular and intraglomerular nerve endings per NEB or glomus cell group, respectively, was measured and the area percentage of mitochondria and synaptic vesicles was determined. In the NEB, no significant difference in total sectional area of the nerve endings between the age groups was observed, while in the carotid body there was a significant increase in the adult age group. In addition, the area percentage of mitochondria and synaptic vesicles of the nerve endings did not change significantly with age in the NEB, while in the carotid body these increased and decreased, respectively, with age. These observations point to a shift from morphologically efferent nerve endings, rich in synaptic vesicles, to morphologically afferent nerve endings, rich in mitochondria. Our interpretation of these findings is that, at birth, the NEB innervation is more mature than the carotid body glomus cell innervation and that the latter matures at a later time than the former. These findings support the theory that the NEB may act as complementary chemoreceptors to the carotid body during the early postnatal period.     

CHOLINE: SELECTIVE ACCUMULATION BY CENTRAL CHOLINERGIC NEURONS

Abstract— Most of the cholinergic input to the hippocampus was destroyed by placement of lesions in the medial septal area. In animals with such lesions we found that hippocampal ChAc activity was reduced by 85–90% and endogenous acetylcholine levels were reduced by more than 80 %. When hippocampal synaptosomes from animals with lesions were incubated with [³H]choline at concentrations of 7.5 nm, 1 μm and 10 μm there was approximately a 60 % reduction in the uptake of [³H]choline, suggesting that cholinergic nerve endings were mainly responsible for [³H]choline uptake. At 0.1 mm concentrations of [³H]choline, there was only a 25 % reduction of choline uptake, suggesting that at higher concentrations of choline there was more nonspecific uptake. The uptake of radiolabelled tryptophan, glutamate and GABA were only slightly or not at all affected by the lesions. There was a significant reduction of uptake of radiolabelled serotonin and norepinephrine, since known monoaminergic tracts were disrupted. Choline uptake was reduced only in brain regions in which cholinergic input was interrupted (i.e. the cerebral cortex and hippocampus) and remained unchanged in other regions (i.e. the cerebellum and striatum). The time course of the reduction in choline uptake was similar to that of the reductions in ChAc activity and endogenous ACh levels; there was no decrease at 1 day, a significant decrease at 2 days, and the maximal decrease at 4 days postlesion. There was a close correlation among choline uptake, ChAc activity and ACh levels in the four brain regions examined (i.e. the striatum, cerebral cortex, hippocampus and cerebellum). Our results suggest that when hippocampal synaptosomes (and perhaps synaptosomes from other brain areas as well) are incubated in the presence of choline, at concentrations of 10 μm m or lower, then cholinergic nerve endings are responsible for the bulk of the choline accumulated by the tissue.     

Distribution of intracardiac neurones and nerve terminals that contain a marker for nitric oxide,NADPH-diaphorase,in the guinea-pig heart

There is strong evidence that NADPH-diaphorase can be used as a marker for neurones that employ nitric oxide as a messenger molecule. In the present study, the NADPH-diaphorase activity of intracardiac neurones and nerve terminals in whole-mount stretch preparations and sections of the newborn and adult guinea-pig atria and interatrial septum has been examined histochemically. Together with epicardial, endothelial and endocardial cells, which displayed some NADPH-diaphorase staining, a subpopulation of intracardiac neurones exhibited moderate-heavy labelling for NADPH-diaphorase, while the majority of neurones were only lightly stained or negative. Intracardiac ganglia containing positive neuronal cell bodies were located between the epicardial cells and atrial myocytes in four main regions: in association with the superior and inferior vena cavae, the points of entry of the pulmonary veins, and within the interatrial septum. Nerve terminals exhibiting NADPH-diaphorase activity were seen throughout the atrial tissue, forming basket-like endings around intracardiac neuronal cell bodies; varicose terminals were also observed on atrial myocytes and other non-neuronal structures. A proportion of the nerve fibres was clearly of intrinsic origin, other terminals may well have originated from neuronal cell bodies present outside the heart.     

High-resolution imaging demonstrates dynein-based vesicular transport of activated Trk receptors

Target-derived neurotrophins signal from nerve endings to the cell body to influence cellular and nuclear responses. The retrograde signal is conveyed by neurotrophin receptors (Trks) themselves. To accomplish this, activated Trks may physically relocalize from nerve endings to the cell bodies. However, alternative signaling mechanisms may also be used. To identify the vehicle wherein the activated Trks are located and transported, and to identify associated motor proteins that would facilitate transport, we use activation-state specific antibodies in concert with immunoelectron microscopy and deconvolution microscopy. We show that the'activated Trks within rat sciatic nerve axons are preferentially localized to coated and uncoated vesicles. These vesicles are moving in a retrograde direction and so accumulate distal to a ligation site. The P-Trk containing vesicles, in turn, colocalize with dynein components, and not with kinesins. Collectively, these results indicate activated Trk within axons travel in vesicles and dynein is the motor that drives these vesicles towards the cell bodies.     

Calcium channel activity in rat brain synaptosomes: Effects of neuroleptics and other factors regulating phosphorylation and transmitter release

Neuroleptic drugs inhibit depolarization-induced Ca uptake in nerve endings, having IC₅₀ values in the micromolar range. Dopamine and a variety of other substances including opiates and PGE₁ are inactive. The effect is probably not mediated by the interaction of the neuroleptics with calmodulin, which itself is a potent inhibitor of stimulated Ca uptake. Dibutyryl cyclic AMP, but not fluoride, increases K⁺-stimulated Ca uptake. Phosphatidic acid, which is an intermediate in transmitter-stimulated phosphatidylinositol turnover, acts as a Ca ionophore in nerve endings and enhances K⁺-stimulated Ca uptake at a relatively low concentration. Carbamyl choline, a known stimulator of phosphatidylinositol turnover, did not, however, cause a significant increase in K⁺-stimulated Ca uptake. Treatment of the nerve ending fraction with relatively small amounts of phospholipase A₂ greatly inhibited depolarization-induced Ca uptake, demonstrating the importance of phospholipids for the functioning of the potential-dependent Ca channel in nerve endings. These studies suggest that the regulation of voltagesensitive Ca channels in nerve endings may be one mechanism controlling transmitter release.     

Identification of Different Types of Spinal Afferent Nerve Endings That Encode Noxious and Innocuous Stimuli in the Large Intestine Using a Novel Anterograde Tracing Technique

In mammals, sensory stimuli in visceral organs, including those that underlie pain perception, are detected by spinal afferent neurons, whose cell bodies lie in dorsal root ganglia (DRG). One of the major challenges in visceral organs has been how to identify the different types of nerve endings of spinal afferents that transduce sensory stimuli into action potentials. The reason why spinal afferent nerve endings have been so challenging to identify is because no techniques have been available, until now, that can selectively label only spinal afferents, in high resolution. We have utilized an anterograde tracing technique, recently developed in our laboratory, which facilitates selective labeling of only spinal afferent axons and their nerve endings in visceral organs. Mice were anesthetized, lumbosacral DRGs surgically exposed, then injected with dextran-amine. Seven days post-surgery, the large intestine was removed. The characteristics of thirteen types of spinal afferent nerve endings were identified in detail. The greatest proportion of nerve endings was in submucosa (32%), circular muscle (25%) and myenteric ganglia (22%). Two morphologically distinct classes innervated myenteric ganglia. These were most commonly a novel class of intraganglionic varicose endings (IGVEs) and occasionally rectal intraganglionic laminar endings (rIGLEs). Three distinct classes of varicose nerve endings were found to innervate the submucosa and circular muscle, while one class innervated internodal strands, blood vessels, crypts of lieberkuhn, the mucosa and the longitudinal muscle. Distinct populations of sensory endings were CGRP-positive. We present the first complete characterization of the different types of spinal afferent nerve endings in a mammalian visceral organ. The findings reveal an unexpectedly complex array of different types of primary afferent endings that innervate specific layers of the large intestine. Some of the novel classes of nerve endings identified must underlie the transduction of noxious and/or innocuous stimuli from the large intestine.     

Neurofilament Proteins Are Synthesized in Nerve Endings from Squid Brain

Abstract: It is generally believed that the proteins of the nerve endings are synthesized on perikaryal polysomes and are eventually delivered to the presynaptic domain by axoplasmic flow. At variance with this view, we have reported previously that a synaptosomal fraction from squid brain actively synthesizes proteins whose electrophoretic profile differs substantially from that of the proteins made in nerve cell bodies, axons, or glial cells, i.e., by the possible contaminants of the synaptosomal fraction. Using western analyses and immunoabsorption methods, we report now that (a) the translation products of the squid synaptosomal fraction include neurofilament (NF) proteins and (b) the electrophoretic pattern of the synaptosomal newly synthesized NF proteins is drastically different from that of the IMF proteins synthesized by nerve cell bodies. The latter results exclude the possibility that NF proteins synthesized by the synaptosomal fraction originate in fragments of nerve cell bodies possibly contaminating the synaptosomal fraction. They rather indicate that in squid brain, nerve terminals synthesize NF proteins.     

Regulation of acetylcholine synthesis in presynaptic endings of cholinergic CNS neurons

Data on acetylcholine (ACh) synthesis in nerve cells are summarized and the mechanism of regulation of this process is described. Under conditions of relative rest on moderate synaptic activity the ACh concentration in the compartment of its synthesis in cholinergic nerve endings is probably maintained at a level corresponding to equilibrium of the reaction catalyzed by the enzyme choline-acetyltransferase (CAT). ACh release is followed by its transport from the compartment of synthesis into the compartment of secretion and automatic resynthesis of new ACh, until equilibrium is restored in the compartment of synthesis. At the same time synaptic activity and ACh release promote synthesis of new ACh by the following pathways. First, a fall in the ACh concentration in the nerve endings disinhibits carriers for choline, and facilitates choline transfer from the extracellular fluid into the cell in accordance with the electrochemical gradient. Second, hydrolysis of liberated ACh increases the choline concentration in the extracellular fluid in the neighborhood of the nerve endings. Third, postactivation hyperpolarization of the nerve endings facilitates transport of choline and an increase in its concentration in the nerve endings. Fourth, there are grounds for considering that stimulation of muscarine receptors promotes a further increase in the choline concentration in the region of the nerve endings by intensification of phosphatidylcholine hydrolysis in postsynaptic cells. Fifth, a decrease in the acetyl-CoA content on account of ACh resynthesis increases pyruvate dehydrogenase activity and acetyl-CoA production. Sixth, it is possible that an increase in the Ca⁺⁺ concentration in nerve endings promotes direct transport of acetyl-CoA from the mitochondria into the cytosol of nerve endings, where ACh is synthesized. It is postulated that under conditions of intensive synaptic activity the rate of supply of acetyl-CoA and choline and also CAT activity in the nerve endings may be factors limiting the velocity of ACh resynthesis.Institute of Physiology, Czechoslovak Academy of Sciences, Prague. Translated from Neirofiziologiya, Vol. 16, No. 5, pp. 603–611, September–October, 1984.     

Differential pre- and postsynaptic effects of desipramine on cardiac sympathetic nerve terminals in RHF

Right heart failure (RHF) is characterized by chamber-specific reductions of myocardial norepinephrine (NE) reuptake, beta-receptor density, and profiles of cardiac sympathetic nerve ending neurotransmitters. To study the functional linkage between NE uptake and the pre- and postsynaptic changes, we administered desipramine (225 mg/day), a NE uptake inhibitor, to dogs with RHF produced by tricuspid avulsion and progressive pulmonary constriction or sham-operated dogs for 6 wk. Animals receiving no desipramine were studied as controls. We measured myocardial NE uptake activity using [(3)H]NE, beta-receptor density by [(125)I]iodocyanopindolol, inotropic responses to dobutamine, and noradrenergic terminal neurotransmitter profiles by glyoxylic acid-induced histofluorescence for catecholamines, and immunocytochemical staining for tyrosine hydroxylase and neuropeptide Y. Desipramine decreased myocardial NE uptake activity and had no effect on the resting hemodynamics in both RHF and sham animals but decreased myocardial beta-adrenoceptor density and beta-adrenergic inotropic responses in both ventricles of the RHF animals. However, desipramine treatment prevented the reduction of sympathetic neurotransmitter profiles in the failing heart. Our results indicate that NE uptake inhibition facilitates the reduction of myocardial beta-adrenoceptor density and beta-adrenergic subsensitivity in RHF, probably by increasing interstitial NE concentrations, but protects the cardiac noradrenergic nerve endings from damage, probably via blockade of NE-derived neurotoxic metabolites into the nerve endings.     

The sphincter of the efferent filament artery in teleost gills: I. Structure and parasympathetic innervation

Previous studies have shown the existence of a sphincter in the efferent filament artery of the teleost gill and its constrictory response to acetylcholine (ACH) and vagal stimulation. This study deals with the muscular organization of this sphincter and the distribution of its innervation as elucidated by degeneration methods and cytochemistry. The sphincter innervation is supplied by the protrematic vagus nerves. Nerve endings filled with cholinergic-type vesicles are located in close association with the adventitial smooth muscle cells and display a strong acetylcholinesterase (ACHE) activity. Section of the protrematic vagus nerve induces a nearly complete degeneration of the sphincter innervation. ACHE-positive nerve cell bodies are present both in the sphincter area and in the protrematic vagus nerve. These results suggest that innervation of the sphincter in the efferent filament artery is cholinergic through the activity of postganglionic axons of the parasympathetic system.     

Regulation of release processes in central serotoninergic neurons

Different technical, physiological and biochemical aspects concerning the study of the release of 5-HT are discussed herein. Isotopic methods are the most suitable techniques since these allow the release of 3H-5-HT to be measured after having determined the identity of the labelled compounds formed from 3H-tryptophan by co-chromatography. Under these conditions, the 3H-amine released in the superfusates comes from serotoninergic nerve endings, since tryptophan hydroxylase is exclusively localized in serotoninergic neurons. Moreover, it appears that newly synthesized 5-HT is preferentially released. The release of 5-HT is dependent on neuronal activity, but is not always linked to the synthesis of 5-HT. The increase in the firing rate of serotoninergic cell bodies by a local application of glutamate in the area of the nucleus raphe dorsalis induces a marked increase n the release of 5-HT in the caudate nucleus; an opposite effect is observed after cooling this region. The local depolarization of serotoninergic terminals located in the caudate nucleus increases the release of this amine. This effect is blocked by TTX. LSD reduces the stimulating effect of KCl, thus indicating that the release of 5-HT can be controlled at a presynaptic level. In addition, the release of the amine is dependent on the presence of calcium. Serotoninergic neuronal activity can be controlled at the preterminal or at the cell body levels by the activity of other neuronal systems. The effects of the release of dopamine from dendrites, and that of GABA in the substantia nigra are reported herein. Furthermore, changes in the activity of the dopaminergic, gabaergic and serotoninergic systems innervating the nucleus raphe dorsalis modulate the release of 5-HT, measured both in the caudate nucleus and in the nucleus raphe magnus. Finally, it has been reported that the release of 5-HT can be estimated in the raphe nuclei dorsalis and magnus. It has been shown that the amounts of 3H-5-HT continuously formed from 3H-TRP and released in the nucleus raphe dorsalis are much greater than those estimated in the caudate nucleus or in the substantia nigra. Although the quantities of endogenous 5-HT measured in the nucleus raphe dorsalis are the highest in the brain, this structure presents only a few serotoninergic nerve endings. This raises the question of the origin of the 5-HT released in serotoninergic nuclei. A possible dendritic release of 5-HT is discussed.