The role of serotonin release and autoreceptors in the dorsalis raphe nucleus in the control of serotonin release in the cat caudate nucleus

Using a push-pull cannula technique and an isotopic method for estimating [3H]serotonin continuously synthesized from [3H]tryptophan, the effects of changes in the release of serotonin in the dorsalis raphe nucleus on in vivo release of [3H]serotonin in the cat caudate nucleus were investigated. The increase in the release of serotonin in the dorsalis raphe nucleus caused by local application of parachlorophenylethylamine (10(-6) M) reduced striatal [3H]serotonin release. This inhibition in serotonin release in the striatum was blocked by the prior and continuous local superfusion of the dorsal raphe with methiothepin (10(-6) M), a serotonin autoreceptor antagonist. GABA (5 x 10(-5) M) applied to the dorsalis raphe reduced both local and striatal release of [3H]serotonin. However, picrotoxin (10(-5) M), a GABA A receptor antagonist applied locally in the dorsalis raphe nucleus increased [3H]serotonin release while decreasing striatal [3H]serotonin release. This decrease in serotonin release in the striatum was again blocked by continuous superfusion of the raphe with methiothepin. Furthermore, superfusion of serotonergic cell bodies of the dorsalis raphe nucleus with methiothepin alone never altered local release or striatal release of [3H]serotonin. These data strongly suggest that the release of serotonin from the cell body in the dorsalis raphe nucleus phasically controls release of the amine at the axonal nerve ending through serotonergic autoreceptors located on serotonergic nerve cell bodies in the dorsalis raphe nucleus. The origin of the serotonin released in the dorsalis raphe nucleus and the possibility that this type of regulation could be related to changes in nerve impulse conduction of the serotonergic raphe-striatal system are discussed.     

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.     

Effects of unilateral electrical stimulation of various thalamic nuclei on the release of dopamine from dendrites and nerve terminals of neurons of the two nigrostriatal dopaminergic pathways

The role of several motor and intralaminar thalamic nuclei in the regulation of dopamine release from terminals and dendrites of the nigrostriatal dopaminergic neurons was investigated in halothane-anaesthetized cats. For this purpose, the effects of the unilateral electrical stimulation of various thalamic nuclei on the release of newly synthesized [3H]dopamine were simultaneously determined in both substantiae nigrae and caudate nuclei using the push-pull cannula method. The electrical stimulation of the motor nuclei was the only one to induce asymmetric changes in the four structures since [3H]dopamine release was enhanced in the ipsilateral caudate nucleus and reduced in the contralateral structure while opposite responses were observed in the corresponding substantiae nigrae. A reduction of [3H]dopamine release occurred in the four structures or only in the contralateral substantia nigra and caudate nucleus following the stimulation of the parafascicularis nucleus and the adjacent posterior part of the nucleus centrum medianum or of the nucleus centralis lateralis and the adjacent paralaminar part of the nucleus medialis dorsalis, respectively. The stimulation of the anterior part of the nucleus centrum medianum, which in contrast to other thalamic nuclei examined, receives few nigral inputs, selectively enhanced [3H]dopamine release in the contralateral substantia nigra. No significant changes in [3H]dopamine release were seen either in the substantiae nigrae or in the caudate nuclei following the stimulation of midline thalamic nuclei. These results indicate that the motor and intralaminar thalamic nuclei exert multiple and selective influences on the release of dopamine from terminals and/or dendrites of the dopaminergic neurons. They also further support a role of thalamic nuclei in the transfer of information from one substantia nigra to the contralateral dopaminergic neurons. The possible involvement of connections between paired thalamic nuclei was underlined by the observations of evoked potentials in contralateral homologous nuclei following unilateral stimulation of motor, or some intralaminar, nuclei. The present report provides new insights on the mechanisms contributing to the reciprocal and/or bilateral regulations of nigrostriatal dopaminergic pathways.     

Origin and influence of the serotoninergic innervation of the subcommissural organ in the rat

The origin of the serotoninergic innervation of the rat subcommissural organ was studied using radioautography of tritiated serotonin and biochemical determination of endogenous serotonin content after electrolytic lesions of raphe nuclei. The results suggest that this innervation is mainly derived from nuclei raphe centralis superior and raphe dorsalis, each nucleus contributing about one-third of the input. A possible contribution from nucleus raphe pontis is also suggested. Given the different patterns of innervation revealed by silver staining of nerve fibers and the different patterns of secretory activity observed with histochemical methods after the electrolytic lesions, the following working hypothesis is formulated. Nucleus raphe dorsalis would inhibit the synthesis of secretory material in the rat subcommissural organ via medium-sized serotoninergic fibers restricted to the hypendymal region, whereas nucleus raphe centralis superior might inhibit the release of secretory material via rather thin serotoninergic fibers reaching the nuclear level of the ependyma. This hypothesis is in line with the inhibitory effect postulated for the serotoninergic innervation in the rat subcommissural organ in early investigations using serotonin neurotoxins.     

Central serotonergic projections to the nucleus tractus solitarii: evidence from a double labeling study in the rat

Projections from several brainstem serotonergic nuclei to the nucleus tractus solitarii were investigated in the rat. Experiments were performed using a double labeling method combining retrograde radioautographic tracing and serotonin immunohistochemistry. After injection of the radioactive tracer ([3H] wheat germ agglutinin) into the lateral nucleus tractus solitarii, nerve cell bodies exhibiting both radioautographic labeling and immunostaining were detected in all the serotonergic nuclei investigated, namely the nucleus raphe magnus, the ventromedial paragigantocellular nucleus, the nuclei raphe pontis, medianus and dorsalis, the medial lemniscus and the reticulotegmental nucleus of the pons. Most of the double labeled perikarya observed were in the nucleus raphe magnus, the adjacent part of the paragigantocellular nucleus and the nucleus raphe dorsalis. Nerve cell bodies retrogradely labeled but devoid of immunostaining were also observed, together with the double labeled perikarya, within serotonergic nuclei. These results provide direct evidence that brainstem serotonergic neurons contribute to the innervation of the nucleus tractus solitarii. They indicate that the nucleus raphe magnus and the nucleus raphe dorsalis constitute two major sources of central serotonergic projections to the nucleus tractus solitarii.     

The source of the transient serotoninergic input to the developing visual and somatosensory cortices in rat.

For approximately the first two weeks of life, dense serotonin immunoreactivity closely matches the pattern of thalamocortical axons innervating both the granular portion of the primary somatosensory cortex and area 17 in rodents [D'Amato et al. (1987) Proc. natn. Acad. Sci. 84, 4322-4326; Fujimiya et al. (1986) J. comp. Neurol. (1986) 246, 191-201; Rhoades et al. (1990) J. comp. Neurol. 293, 190-207]. This serotonin immunoreactivity is not contained in thalamocortical axons [Rhoades et al. (1990) 293, 190-207] but its source has never been demonstrated. In the present study, a variety of approaches were used to address this issue. The combination of electron microscopy and immunocytochemistry showed that all serotonin immunoreactivity in the developing cerebral cortex was contained in axons and that the terminals of many of these fibers made synapses with the dendrites of cortical cells. Treatment with fluoxetine, a specific inhibitor of serotonin uptake, did not result in a loss of the cortical pattern of serotonin immunoreactivity, indicating that immunoreactive fibers were not labeled solely as a result of serotonin uptake. The combination of retrograde tracing from the primary somatosensory cortex and area 17 with immunocytochemistry demonstrated numerous double-labeled cells in nucleus raphe dorsalis and the median raphe nucleus. Smaller numbers of double-labeled neurons were located in the B9 cell group and the region of the lateral midbrain tegmentum. Large electrolytic lesions that included most of the nucleus raphe dorsalis and median raphe nucleus, but which left the B9 group and more caudal serotoninergic cells undamaged, caused either a substantial reduction in density or complete disappearance of the serotonin pattern in both hemispheres. Unilateral electrolytic lesions of the medial forebrain bundle resulted in a loss of the pattern only on the side of the damage. Injection of the neurotoxin 5,7-dihydroxytryptamine directly into the mesencephalon either abolished or substantially reduced the density of the cortical serotonin immunoreactivity. Injections that produced substantial cell loss in the median raphe nucleus, but only minor cell loss in the nucleus raphe dorsalis had little effect upon the cortical pattern of serotonin immunoreactivity. These results indicate that the dense serotonin immunoreactivity which appears transiently in the visual and somatosensory cortices of perinatal rodents is contained in serotoninergic axons that arise from cells in the nucleus raphe dorsalis and perhaps also the median raphe nucleus.     

Substance P and neurokinin A regulate by different mechanisms dopamine release from dendrites and nerve terminals of the nigrostriatal dopaminergic neurons

Numerous striatal neurons innervating the substantia nigra contain substance P and/or neurokinin A. In contrast to substance P or neurokinin A, little neurokinin B is found in the substantia nigra. This led us to compare the effects of nigral application of these tachykinins on the release of dopamine from dendrites and nerve terminals of nigrostriatal dopaminergic neurons. Experiments were made in halothane-anesthetized cats implanted with one push-pull cannula in the substantia nigra and another in the ipsilateral caudate nucleus [3H]Tyrosine was delivered continuously to each push-pull cannula and the release of newly synthesized [3H]dopamine measured in the superfusate. Unlike substance P or neurokinin A, neurokinin B (10(-8) M) applied for 30 min into the pars compacta of the substantia nigra was without effect on the release of [3H]dopamine from nerve terminals or dendrites. When either substance P (10(-8) M) or neurokinin A (10(-8) M) was applied into the pars compacta, the release of [3H]dopamine from nerve terminals was enhanced. While neurokinin A also stimulated the dendritic release of [3H]dopamine, this was reduced by substance P. At a lower concentration (10(-9) M), neurokinin A induced similar effects to those observed at 10(-8) M whereas substance P (10(-9) M) stimulated moderately [3H]dopamine release from nerve terminals but did not affect the dendritic release of the [3H]amine. When superfused into the pars reticulata, substance P (10(-8) M) still stimulated [3H]dopamine release from nerve terminals but not from dendrites while neurokinin A (10(-8) M) was without effect either in the caudate nucleus or the substantia nigra. Additional experiments were made to determine whether or not substance P (10(-8) M) or neurokinin A (10(-8) M) act directly on nigral dopaminergic neurons when applied into the pars compacta. The effects of substance P on [3H]dopamine release from nerve terminals and dendrites were prevented when 2-amino-6-trifluoromethoxy benzothiazole (10(-5) M), an antagonist of glutamatergic transmission, was applied continuously into the caudate nucleus. In contrast, the stimulatory effects of neurokinin A on [3H]dopamine release from nerve terminals and dendrites were insensitive to 2-amino-6-trifluoromethoxy benzothiazole (10(-5) M). These results suggest that neurokinin A, but not substance P, acts directly on dopaminergic cells. In the light of previous observations, we propose that the effects of substance P on dopaminergic transmission are mediated by a nigro-thalamo-cortico-striatal loop.(ABSTRACT TRUNCATED AT 400 WORDS)     

Forebrain and brainstem afferents to the arcuate nucleus in the rat: potential pathways for the modulation of hypophyseal secretions

Doxorubicin, an anti-oncogenic agent, was used as a retrograde marker to identify arcuate nucleus afferent projections. Injections of this tracer into the arcuate nucleus indicated that the subfornical organ, the organum vasculosum of the lamina terminalis, the nucleus raphe dorsalis and median raphe send projections to the arcuate nucleus. Immunocytochemical procedures were used to demonstrate that the raphe projections to the arcuate nucleus are serotoninergic. This anatomical investigation provides evidence that neural pathways exist between forebrain body fluid and mineral nuclei, mesencephalic serotonin nuclei and the arcuate nuclei.     

Different effects of electrical stimulation of the mesencephalic and pontine reticular formation on the release of dopamine and acetylcholine in the cat caudate nucleus

The effects of unilateral electrical stimulation of the pontine (PRF) and mesencephalic (MRF) reticular formation on the release of acetylcholine (ACh) and of [3H]dopamine continuously synthesised from [3H]tyrosine were examined in both caudate nuclei of halothane-anaesthetised cats implanted with push-pull cannulae. Stimulation of PRF led to a prolonged bilateral increase in the release of [3H]dopamine, whereas a significant reduction in [3H]amine release was observed in the ipsilateral caudate nucleus following stimulation of the MRF. Changes in ACh release were also seen, but they seemed to be independent from those in dopamine release: the release of ACh was enhanced markedly in both caudate nuclei following stimulation of the MRF, whereas a more moderate increase in the release of ACh occurred ipsilaterally following stimulation of the PRF. These data indicate that both the MRF and the PRF are involved in the control of dopaminergic and cholinergic transmission in the basal ganglia.     

Are presynaptic dopamine autoreceptors and postsynaptic dopamine receptors in the rabbit caudate nucleus pharmacologically different?

Slices of the rabbit caudate nucleus were preincubated with [3H]dopamine or [3H]choline and then superfused and stimulated electrically. Apomorphine reduced the stimulation-evoked overflow of tritium over the same concentration range, independently of whether slices had been pre-incubated with [3H]dopamine or with [3H]choline. Each of three antagonists--molindone, sulpiride and metoclopramide--increased the evoked overflow of tritium over the same concentration range in experiments with [3H]dopamine and those with [3H]choline. For each antagonist, the pA2 values against apomorphine obtained in [3H]dopamine experiments and in [3H]choline experiments were very similar. This study is a functional in vitro approach to receptor characterization, as opposed to radioligand binding studies or in vivo investigations. The results show that the dopamine receptor agonist apomorphine and three antagonists are unable to distinguish between the presynaptic, release-inhibiting dopamine autoreceptors and those postsynaptic dopamine receptors which, when activated, depress the release of acetylcholine. Although there are certainly more dopamine receptors in the caudate nucleus, these two physiologically important groups seem to be closely related.     

Reduced high-affinity glutamate uptake sites in the brains of patients with Huntington's disease

The binding of D-[3H]aspartic acid to the high-affinity glutamate uptake system was studied in membrane preparations of postmortem brains from controls and Huntington's disease (HD) subjects. The groups were matched for age and postmortem delay. A large (60-72%) and significant reduction in D-[3H]aspartate binding was observed in both the caudate nucleus and putamen, but not in the frontal cortex of the HD brains. The loss of striatal D-[3H]aspartate binding may reflect a loss of the high-affinity glutamate uptake system contained on the terminals of corticostriatal afferents. In contrast, the binding of [3H]paroxetine to the serotonin uptake system was marginally increased in the caudate nucleus and unchanged in the putamen. It is suggested that the reduction of high-affinity glutamate uptake sites may contribute to the production of the striatal lesion in HD.     

大鼠孤束核内5－羟色胺能纤维的来源──HRP逆行追踪与5－羟色胺免疫组织化学双标记洁研究

用ＨＲＰ注入孤束核逆行追踪和５－羟色胺（５－ＨＴ）免疫组织化学相结合的双标记法，观察大鼠孤束核内的５－ＨＴ能纤维和终末的来源。结果在脑干内见到许多ＨＲＰ单标、５－ＨＴ单标和ＨＲＰ／５－ＨＴ双标细胞，其中ＨＲＰ／５－ＨＴ双标细胞主要分布于脑桥被盖核（２６．５７％）、脑桥网状核（２４．１９％）、中缝大核（２１．１７％），其次为中缝隐核（１２．７４％）、中缝苍白核（７．７８％），少数见于中缝背核、中脑导水管周围灰质和中缝脑桥核等处。而在结状神经节内仅见到ＨＲＰ单标细胞，未见到５－ＨＴ单标和ＨＲＰ／５－ＨＴ双标细胞。以上结果表明，大鼠孤束核的５－ＨＴ能纤维和终末主要来源于脑桥与延髓的５－ＨＴ神经元的投射。本研究提示脑桥与延髓内上述核团的５－ＨＴ神经元通过向孤束核的直接投射参与孤束核内脏功能的调控。     

Ultrastructural relationships between serotonin and dopamine neurons in the rat arcuate nucleus and medial zona incerta: a combined radioautographic and immunocytochemical study

Combined radioautographic and immunocytochemical detection of [3H]serotonin-labeled axon terminals and tyrosine hydroxylase-immunoreactive processes in the same thin sections allowed for electron microscopic demonstration of direct appositions between serotoninergic axonal varicosities and dopaminergic nerve cell bodies and/or dendrites in the anterior part of the arcuate nucleus and in the medial zona incerta. Although no junctional specializations were apparent at the sites of contacts, it is proposed that the observed appositions may represent a serotonin input onto tubero-infundibular and incerto-hypothalamic dopaminergic neurons. This innervation could account for some of the central neuroendocrine effects of serotonin, particularly its regulatory role on prolactin and gonadotropin secretion.     

In vivo presynaptic control of dopamine release in the cat caudate nucleus--II. Facilitatory or inhibitory influence of L-glutamate

The local effects of various concentrations of L-glutamate (from 10(-8) M up to 10(-3) M) on the release of [3H]dopamine synthesized continuously from [3H]tyrosine were examined in the caudate nucleus of halothane-anaesthetized cats implanted with push-pull cannulae. When used at a concentration of 10(-8) M or 10(-7) M, L-glutamate stimulated the release of [3H]dopamine from nerve terminals of the nigrostriatal dopamine neurons. This effect was still observed in the presence of tetrodotoxin (5 X 10(-7) M) but it was antagonized by 2-amino 6-trifluoromethoxy benzothiazole (PK 26124) (10(-5) M), an antagonist dopamine nerve terminals. While no significant change in the release of [3H]dopamine was observed with 10(-6) M L-glutamate, higher concentrations (from 10(-5) M to 10(-3) M) of the amino acid produced a long-lasting reduction in the [3H]transmitter release. This latter effect was also antagonized by PK 26124 (10(-5) M) but, unlike that observed with 10(-8) M L-glutamate, it did not persist in the presence of tetrodotoxin (5 X 10(-7) M). On the contrary, a marked stimulation of the release of [3H]dopamine was seen in the presence of this neurotoxin. The reduction in the release of [3H]dopamine produced by 10(-4) M L-glutamate was also antagonized by bicuculline (10(-5) M) and moreover a marked stimulation of [3H]dopamine release took place in the presence of this gamma-aminobutyric acid (GABA) antagonist. Therefore, high concentrations of L-glutamate exerted an inhibitory presynaptic control on [3H]dopamine release which seemed to be indirect and mediated partly by GABAergic neurons. Since a sustained reduction in the spontaneous release of [3H]dopamine was seen in the presence of PK 26124, the corticostriatal glutamatergic neurons appeared to exert a tonic facilitatory presynaptic influence on dopamine release. This effect was important since it represented 40% of the tetrodotoxin-sensitive release of the [3H]transmitter. The direct (stimulatory) and indirect (inhibitory) presynaptic controls on dopamine release mediated by corticostriatal glutamatergic fibres are discussed in light of previous findings and of the anatomical organization of the caudate nucleus.     

gamma-Aminobutyric acid and 5-hydroxytryptamine interrelationship in the rat nucleus raphe dorsalis: combination of radioautographic and immunocytochemical techniques at light and electron microscopy levels

Serotonin and gamma-aminobutyric acid (GABA) neurons in the nucleus raphe dorsalis were identified by immunocytochemistry using antibodies to 5-hydroxytryptamine or GABA. The pattern of the 5-hydroxytryptamine and GABA immunostaining presented similar features: 5-hydroxytryptamine or GABA immunoreactive somata were fusiform or ovoid (15-20 micron) and positive dendritic profiles were found either without any connection with other nerve elements or in contact with one or several terminals. In addition, some 5-hydroxytryptamine nerve endings were apposed to 5-hydroxytryptamine immunoreactive cell bodies or dendrites; also some GABA-immunopositive terminals were in contact with GABA-immunopositive nerve cell bodies. On the other hand, GABA and 5-hydroxytryptamine patterns may be differentiated in several respects: the 5-hydroxytryptamine-reactive nerve cell bodies were more numerous than the GABA ones. Some small, round (8-10 micron) nerve cell bodies were reactive with GABA antiserum, but no neurons of this type were reactive with a 5-hydroxytryptamine antiserum; finally, GABA nerve terminals were more numerous than 5-hydroxytryptamine ones. In order to understand the relationship between GABA and 5-hydroxytryptamine neurons, radioautographic and immunocytochemical procedures were combined: 5-hydroxytryptamine and GABA immunocytochemistry was combined with radioautography of [3H]GABA and [3H]5-hydroxytryptamine uptake, respectively. Some nerve cell bodies, dendrites or terminals, which were 5-hydroxytryptamine-immunopositive, were also capable of accumulating [3H]GABA and, conversely, some GABA-immunopositive elements were capable of accumulating [3H]5-hydroxytryptamine. Moreover, several nerve elements were reactive with both glutamate decarboxylase and 5-hydroxytryptamine antisera. These data confirm in electron microscopy previous studies suggesting the coexistence of both GABA and 5-hydroxytryptamine in the same neurons. The presence of uptake mechanisms for GABA and 5-hydroxytryptamine may indicate the action of both neurotransmitters in the same neuron. On the other hand, the [3H]GABA-labelled nerve endings in contact with 5-hydroxytryptamine-positive dendrites or nerve cell bodies indicate the possibility of a GABAergic control of the activity of some 5-hydroxytryptamine neurons; this corroborates biochemical and electrophysiological studies whereby a trans-synaptic control of the 5-hydroxytryptamine neurons by GABA may be envisaged.     

Functional heterogeneity of the matrix compartment in the cat caudate nucleus as demonstrated by the cholinergic presynaptic regulation of dopamine release.

Previously, using a new in vitro microsuperfusion procedure, we have demonstrated marked differences in the cholinergic presynaptic regulation of the release of [3H]dopamine continuously synthesized from [3H]tyrosine in two close striosomal- and matrix-enriched areas of the cat caudate nucleus. A tetrodotoxin-resistant stimulatory effect of acetylcholine mediated by muscarinic receptors was observed in both compartments. However, in addition, two opposing types of tetrodotoxin-sensitive acetylcholine-evoked regulation of [3H]dopamine release were only seen in the matrix: one facilitatory, involving nicotinic receptors located on as yet unidentified neurons, and the other inhibitory, mediated by muscarinic receptors located on dynorphin-containing neurons. In the present study, using the same approach, a functional heterogeneity was demonstrated in the matrix. Indeed, in various conditions the effects of acetylcholine (50 microM) on the release of [3H]dopamine were different in a matrix-enriched area (matrix 2) distinct from that previously investigated (matrix 1); these areas being characterized by the presence or absence of islands of striatonigral cells, respectively. As in matrix 1, acetylcholine induced a short-lasting stimulation of [3H]dopamine release in matrix 2 but, in contrast to that observed in matrix 1, the acetylcholine-evoked response in matrix 2 was not modified in the presence of tetrodotoxin (1 microM). Experiments made in the presence of the tetrodotoxin and atropine (1 microM) indicated that both muscarinic and nicotinic receptors are located on dopaminergic nerve terminals in matrix 2 while muscarinic receptors are only present in matrix 1. In the absence of tetrodotoxin, the short-lasting stimulation of [3H]dopamine release was transformed into a long-lasting response in the presence of pempidine (50 microM), in matrix 2 but not in matrix 1 while prolonged responses were seen in both matrix areas in the presence of atropine. Finally, the acetylcholine short stimulatory effect on [3H]dopamine release was transformed into a long stimulatory response in the presence of bicuculline (50 microM) but not naloxone (1 microM) in matrix 2 while the reverse was observed in matrix 1. By providing further evidence for a functional heterogeneity of the matrix, our results suggest that depending on the matrix area investigated, dynorphin- or GABA-containing neurons are involved in the indirect cholinergic inhibitory control of dopamine release.     

Cholinergic and noradrenergic denervations decrease labelled purine release from electrically stimulated rat cortical slices

The origin of cortical purine release was investigated by measuring [3H]purine release from electrically stimulated cortical slices of rats after neurotoxic lesions of cholinergic, noradrenergic and serotoninergic pathways innervating the cortex. Purines were labelled by incubating the cortical slices with [3H]adenine. The 3H efflux at rest and during stimulation, analysed by high performance liquid chromatography, consisted of adenosine, inosine, hypoxanthine and a small amount of nucleotides. Twenty days after unilateral or bilateral lesion of the nucleus basalis a marked decrease in choline acetyltransferase activity was associated with a decrease in [3H]purine release. A linear relationship was found between the decrease in choline acetyltransferase activity and [3H]purine release. A partial recovery in both choline acetyltransferase activity and [3H]purine release was observed eight months after the lesion. Twenty days after intra-cerebroventricular injection of 6-hydroxydopamine a 59% decrease in cortical noradrenaline content was associated with a 44% decrease in [3H]purine release. Conversely, no change in [3H]purine release was found in rats in which a 89% decrease in cortical serotonin content was induced by intra-cerebroventricular injection of 5,7-dihydroxytryptamine. The decrease in [3H]purine release after the lesion of the cholinergic and noradrenergic pathways may depend on metabolic changes, a loss of a stimulating influence of acetylcholine and noradrenaline or may indicate a release of [3H]purine from cholinergic and noradrenergic fibres.     

The significance of nucleus raphe dorsalis and centralis for thermoafferent signal transmission to the preoptic area of the rat

Summary Neuronal spike-trains were recorded extracellularly within the preoptic area, which is generally considered the center of information processing for thermoregulation. The neuronal responses were tested by thermal stimulation of the scrotal and abdominal skin of the rat. After a neuron had been identified as warm responsive, electrical stimulation and/or lesion were applied in medial midbrain to test the influence on the preoptic neurons. Post-stimulus-histograms were determined by using 600 single shocks. Furthermore a small area of the medial midbrain was electrolytically lesioned and the thermal response of the neuron was tested again. All lesion, stimulation and recording sites were examined histologically. Electrical stimulation of either nucleus raphe dorsalis or centralis influenced the responsiveness of the recorded preoptineurons. The post-stimulus-histograms predominantly reveal polysynaptic ascending pathways. Lesion of either nucleus raphe dorsalis or centralis abolished the responsiveness of the recorded preoptic neurons. Lesions lateral to these nuclei were ineffective. By this it is concluded that both nucleus raphe dorsalis and centralis are essentially involved in the thermal signal transmission to the preoptic area.     

大鼠臂旁核内5-羟色胺阳性纤维和终末的来源

本研究采用荧光金(FG)逆行追踪与5-羟色胺(5-HT)免疫荧光组化染色相结合的双重技术观察了臂旁核(PBN)内5-HT阳性神经纤维和终末的来源。将FG注入PBN后,FG逆标神经元主要分布在三叉神经核簇、脑干网状结构外侧小细胞部、中缝核簇和中脑导水管周围灰质(PAG);免疫荧光组化染色的结果显示5-HT阳性神经元主要位于中缝核簇和PAG;在中缝核簇和PAG内可见FG逆标并呈5-HT阳性的双重标记神经元。上述结果表明,中缝核簇和PAG内的5-HT能神经元向PBN发出投射,它们在躯体和内脏感觉信息的传递和调控方面发挥重要作用。     

Release of endogenous 5-hydroxytryptamine in rat ventral hippocampus evoked by electrical stimulation of the dorsal raphe nucleus as detected by microdialysis: sensitivity to tetrodotoxin, calcium and calcium antagonists

We have utilized the brain microdialysis technique in an attempt to measure excitation-secretion coupled release of endogenous 5-hydroxytryptamine in rat brain in vivo and investigated the pharmacology of the voltage-sensitive calcium channel involved in this process. All experiments were carried out using chloral hydrate anaesthetized rats. Ascending serotoninergic neurons were electrically stimulated using an electrode implanted into the dorsal raphe nucleus. A dialysis probe was implanted into the ventral hippocampus and continuously perfused with artificial cerebrospinal fluid containing the selective 5-hydroxytryptamine uptake inhibitor citalopram (1 microM). Twenty-minute perfusates were analysed for endogenous 5-hydroxytryptamine using high performance liquid chromatography with electrochemical detection. Electrical stimulation (cathodal monophasic 1 ms pulses, 300 microA, 2-10 Hz) of the dorsal raphe nucleus for 20 min induced an immediate release of 5-hydroxytryptamine which lasted for the duration of the stimulus and was frequency-dependent. The calculated amount of 5-hydroxytryptamine release per electrical impulse was constant over the frequency range used. Addition of tetrodotoxin (10 microM) to, or omission of calcium from, the perfusion medium reduced the spontaneous output of 5-hydroxytryptamine by 60-70% and caused a near complete inhibition of the effect of low frequency (3 Hz) electrical stimulation of the dorsal raphe nucleus. Local perfusion with cadmium (30 and 300 microM), which is reported to antagonize both N- and L-type voltage-sensitive calcium channels, also caused a pronounced decrease of basal output of 5-hydroxytryptamine and a marked, but not complete inhibition of the effect of nerve stimulation.(ABSTRACT TRUNCATED AT 250 WORDS)