The Cerebral Cortex and Parafascicular Thalamic Nucleus Facilitate In vivo Acetylcholine Release in the Rat Striatum through Distinct Glutamate Receptor Subtypes

Electrical stimulation (ten pulses of 0.5 ms, 10 V applied over 10 s at 10 Hz, 140 pA) delivered bilaterally to the prefrontal cortex or the parafascicular thalamic nucleus of freely moving rats facilitated acetylcholine release in dorsal striata, assessed by trans-striatal microdialysis. The facilitatory effects were blocked by coperfusion with 5 μM tetrodotoxin, suggesting that the release was of neuronal origin. The response of the striatal cholinergic neurons to prefrontal cortical stimulation was short-lived and required a longer period of stimulation (20 min) than the response to thalamic stimulation (4 min) to reach maximal effect. The α-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA)/kainate glutamatergic receptor antagonist 6,7-dinitroquinoxaline-2,3-dione [DNQX; 12 nmol per side, intracerebroventricularly (i.c.v.)] and the AMPA antagonist 6-nitro-7-sulphamoylbenzo(f)quinoxaline-2,3-dione (NBQX; 12 nmol per side, i.c.v. or 12.8 μM infused into the striatum), but not the NMDA-type receptor antagonist MK-801 (0.2 mg/kg, i.p.), abolished the facilitatory effect on striatal acetylcholine release evoked by stimulation of the prefrontal cortex. By contrast, DNQX or NBQX did not prevent the increase in striatal acetylcholine release evoked by parafascicular nucleus stimulation, but MK-801, in accordance with previous results, did so. MK-801 by itself lowered striatal acetylcholine output while DNQX and NBQX did not. The results provide in vivo evidence that the cerebral cortex facilitates cholinergic activity in the dorsal striatum apparently through the non-tonic activation of AMPA-type glutamatergic receptors while the parafascicular nucleus does this through tonic activation of NMDA receptors. Both glutamate receptor types are probably located in the striatum. The overall results suggest that the two pathways operate independently to regulate striatal cholinergic activity through distinct mechanisms.     

Dopamine efflux in the rat striatum evoked by electrical stimulation of the subthalamic nucleus: potential mechanism of action in Parkinson's disease

The precise mechanism whereby continuous high-frequency electrical stimulation of the subthalamic nucleus ameliorates motor symptoms of Parkinson's disease is unknown. We examined the effects of high-frequency stimulation of regions dorsal to and within the subthalamic nucleus on dopamine efflux in the striatum of urethane-anaesthetized rats using constant potential amperometry. Complementary extracellular electrophysiological studies determined the activity of subthalamic nucleus neurons in response to similar electrical stimulation of the subthalamic nucleus. High-frequency stimulation of the subthalamic nucleus increased action potential firing in the subthalamic nucleus only during the initial stimulation period and was followed by a cessation of firing over the remainder of stimulation. Electrical stimulation of the subthalamic nucleus with 15 pulses elicited stimulus-time-locked increases in striatal dopamine efflux with maximal peak effects occurring at 50 Hz frequency and 300 microA intensity. Extended subthalamic nucleus stimulation (1000 pulses at 50 Hz; 300 microA) elicited a similar peak increase in striatal dopamine efflux that was followed by a relatively lower steady-state elevation in extracellular dopamine over the course of stimulation. In contrast, extended stimulation immediately adjacent and dorsal to the subthalamic nucleus resulted in an 11-fold greater increase in dopamine efflux that remained elevated over the course of the stimulation. Immunohistochemical staining for tyrosine hydroxylase revealed catecholaminergic fibers running immediately dorsal to and through the subthalamic nucleus. Taken together, these results suggest that enhanced dopamine release within the basal ganglia may be an important mechanism whereby high-frequency stimulation of the subthalamic nucleus improves motor symptoms of Parkinson's disease.     

Regional variations in the physiology of the rat caudate-putamen. 2. Effects of amphetamine and amphetamine induced dopamine release on basal and cortical stimulation evoked multiple unit activity

Summary. Dopaminergic terminals within the caudate-putamen are located in an ideal position to modulate the corticostriatal system. Since this is the major afferent system of the striatum, dopamine has very powerful effects on striatal electrophysiological activity. The striatum is a regionally specialized multifunctional nucleus. It is therefore important to determine if dopamine has the same modulatory effects within different areas of the nucleus. The effects of 2.5 mg/Kg D-amphetamine (IP) on cortical stimulation evoked and basal multiple unit activity (MUA) was measured in 7 dorsal and 7 ventral striatal areas of the urethane anaesthetized rat. In general, amphetamine caused an increase in the basal activity and a decrease in the cortical stimulation evoked activity. However, there were both qualitative and quantitative regionally dependent differences in these responses. The effect on basal MUA was more pronounced in the dorsal and caudal areas whereas the effect on cortical stimulation evoked MUA was more pronounced in the ventral areas. The electrophysiological effects of amphetamine within the striatum were correlated with its regionally dependent effects on extracellular dopamine. This produced a measure of the effects of striatal dopamine on regional electrophysiological activity. This information was also used to determine the mathematical relationship between dopamine concentration change and the change in MUA. These data indicate that the excitatory effects of amphetamine-induced dopamine release on the non-stimulated MUA progressively increase along the rostro-caudal axis of the nucleus. In addition, the effects were more pronounced in the ventromedial as compared to the ventrolateral areas. These effects correlated best with the rate of change in dopamine concentration. In the dorsal striatum amphetamine-induced increases in dopamine had a regionally homogeneous inhibitory effect on the stimulated MUA. In the ventral striatum however, it had a progressively stronger effect along the rostro-caudal axis. These effects correlated best with the absolute change in dopamine concentration. Received April 17, 2002; accepted December 2, 2002 Published online March 5, 2003 Authors' address: Dr. G. Glynn, School of Pharmacy and Allied Health Professions, Creighton University, 2500 California Plaza, Omaha, NE 68178, U.S.A., e-mail: GGlynn@creighton.edu     

Striatal control of locomotion, intentional actions and of integrating and perceptive activity.

The three parts of the striatum — putamen, caudate nucleus and fundus striati — control intentional actions and not simple movements without conscious representation. According to its integrative function the striatum consists of at least four distinct types of nerve cells, the bulk of which (more than 90%) is formed by the small “spiny” neurons with intrinsic axons. In each part of the striatum nine discrete types of synapses are distinguished: one axo-spinous and one axo-dendritic (or axo-somatic) from the substantia nigra (type I and II); from the cortical fields (type III and VII); from the center median-parafascicular complex of the thalamus (type IV and VII) and type V as axon-collateral of the large efferent neurons; type VI with flattened vesicles of so far unknown origin; type VIII as dendritic terminal of Golgi type II cells; and type IX as intrinsic contacts between the small intrinsic and the large efferent neurons. The transmitter of the nigral afferents is dopamine (DA), that of the cortical afferents glutamic acid, that of the intrinsic synapses acetylcholine (ACh) and that of center median-parafascicular afferents also perhaps ACh. The results of the integrating activities of this synaptic organization are conducted by GABA-ergic, mostly inhibitory, fibers to the substantia nigra and to both segments of the pallidum.The strio-pallidal neuronal chain forms a part of the nonspecific thalamo-cortical activating system, which we prefer to call trunco-thalamic system to avoid the equivocal term, nonspecific, and because it is anatomically more correct. The nonspecific input of the different sensory systems to the cortical sensory regions runs through the neuronal chains of the reticular formation to the trunco-thalamic nuclei which do not project directly to the cortical fields as the intralaminar nuclei and mainly the center median and parafascicular nuclei. These trunco-thalamic nuclei project to the three parts of the striatum or to the outer segment of the pallidum. All these impulses meet, finally, in the outer and inner segments of the pallidum. From there at least two, but probably more, pathways go to thalamic nuclei, such as V.o.a. (the anterior basal part of VL) and VA which project directly to cortical fields specifically or diffusely to the rostral part of the hemisphere and to the parietal lobe. This nonspecific action of the basal ganglia loop on the majority of cortical areas is also demonstrated by eliciting recruiting potentials in gross recording and field potential recordings. Each part of the striatum belongs to the trunco-thalamic activity-regulating system of the cerebral cortex.The direct functional role of the putamen is best revealed by the Hess method of near threshold stimulation in the unanesthetized, unrestrained cat: low frequency stimulation (1–9 Hz) produces a delayed inhibition or motor ritardando of any form of spontaneous or stimulus induced movement and a closing of the eyes with miosis, and intermediate frequency stimulation (15–50 Hz) produces an immediate arrest of all activities and opening of the eyes with mydriasis. A locomotor activity, turning to the side of stimulated putamen, is elicited at the same time and a spontaneous turning to this side is accelerated and a spontaneous turning to the nonstimulated side suppressed. In contrast, low and middle frequency pallidal stimulation elicits only excitatory effects with turning to the contralateral side and pupil dilatation; this is true for animal experiments and during stereotaxic operations in the human.From these data it is concluded that the function of the putamen is at the same time to focus the attention, the emotional participation and the excitability on one single event by simultaneously suppressing and fading out all other happenings and motivational objects, including those coming from the contralateral side. The pallidal function is horizontal locomotion to the contralateral side, the enhancing of the muscle tone of the contralateral side and directing of attention to the contralateral side; without this no conscious mental processes can go on.     

Raclopride or high‐frequency stimulation of the subthalamic nucleus stops cocaine‐induced motor stereotypy and restores related alterations in prefrontal basal ganglia circuits

Motor stereotypy is a key symptom of various neurological or neuropsychiatric disorders. Neuroleptics or the promising treatment using deep brain stimulation stops stereotypies but the mechanisms underlying their actions are unclear. In rat, motor stereotypies are linked to an imbalance between prefrontal and sensorimotor cortico‐basal ganglia circuits. Indeed, cortico‐nigral transmission was reduced in the prefrontal but not sensorimotor basal ganglia circuits and dopamine and acetylcholine release was altered in the prefrontal but not sensorimotor territory of the dorsal striatum. Furthermore, cholinergic transmission in the prefrontal territory of the dorsal striatum plays a crucial role in the arrest of motor stereotypy. Here we found that, as previously observed for raclopride, high‐frequency stimulation of the subthalamic nucleus (HFS STN) rapidly stopped cocaine‐induced motor stereotypies in rat. Importantly, raclopride and HFS STN exerted a strong effect on cocaine‐induced alterations in prefrontal basal ganglia circuits. Raclopride restored the cholinergic transmission in the prefrontal territory of the dorsal striatum and the cortico‐nigral information transmissions in the prefrontal basal ganglia circuits. HFS STN also restored the N‐methyl‐d ‐aspartic‐acid‐evoked release of acetylcholine and dopamine in the prefrontal territory of the dorsal striatum. However, in contrast to raclopride, HFS STN did not restore the cortico‐substantia nigra pars reticulata transmissions but exerted strong inhibitory and excitatory effects on neuronal activity in the prefrontal subdivision of the substantia nigra pars reticulata. Thus, both raclopride and HFS STN stop cocaine‐induced motor stereotypy, but exert different effects on the related alterations in the prefrontal basal ganglia circuits.     

Galanin-evoked acetylcholine release in the rat striatum is blocked by the putative galanin antagonist M15

The effect of the putative galanin (GAL) antagonist M15 on the Gal-evoked release of acetylcholine (ACh) was investigated in the rat striatum using microdialysis and HPLC techniques. GAL (3 nmol/10 microliters), applied in the lateral ventricle (i.c.v.) or perfused through the microdialysis membrane into the striatum, was found to enhance basal ACh release. The GAL-evoked release of striatal ACh was completely blocked by M15 (3 nmol/10 microliters), administered either i.c.v. or into the straitum. This finding suggests that GAL stimulates the basal ACh release in the rat striatum by a direct action of the peptide on striatal GAL receptors.     

Effects of thalamic parafascicular stimulation on the periaqueductal gray and adjacent reticular formation neurons. A possible contribution to pain control mechanisms

To investigate the mechanism of analgesic effect of electrical stimulation of the thalamic parafascicular nucleus (Pf), we studied modulations of neuronal activities in the periaqueductal gray (PAG) and the adjacent reticular formation (RF) in response to Pf electrical and peripheral noxious stimulations in the rat. Extracellular single unit activities were recorded from 129 neurons in the PAG and adjacent RF under light halothane anesthesia. Pf stimulation caused neuronal responses in approximately 80% of the PAG and adjacent RF neurons, and noxious stimulation in 75%, with predominant excitatory responses to either stimulation. When the responses to the two stimuli were tested in the same neurons (n = 69), 91% responding to noxious stimuli also responded to Pf stimuli, again with predominant excitatory responses to either stimulation. The PAG and adjacent RF neurons that were verified antidromically to project to the nucleus raphe magnus (NRM), showed a similar pattern of response (n = 20). These results suggest that a sizeable population of neurons in the PAG and adjacent RF receives excitatory effects from the Pf and noxious afferents, and that part of these neurons projects to the NRM, which inhibits the dorsal horn cells of the spinal cord (the descending pain suppression system). Thus, part of the mechanism of the analgesic effects of Pf stimulation is due to activation of the descending pain suppression system by exciting the PAG and adjacent RF neurons. A possible role of noxious afferents on the negative feedback to pain mediation through this descending system also has to be considered.     

The effects of protoveratrine and germines on the release of acetylcholine from the auerbach plexus of the guinea-pig ileum

The effect of protoveratrine A and germine-3-acetate (GMA) on the release mechanism of acetylcholine from the nerve terminals of the Auerbach plexus in the longitudinal muscle layer of the guinea-pig ileum was studied. Protoveratrine A, GMA and germine potentiated neuroeffector transmission in Auerbach's plexus in the longitudinal muscle preparation provided the neurons were stimulated at low frequencies (less than 10 Hz). Protoveratrine A and GMA enhanced the release of acetylcholine from the nerve terminals during the resting period and at low frequency of stimulation (less than 10 Hz). At continous stimulation with high frequency they were ineffective (greater than 10 Hz). When trains of 20 stimuli, with a pulse interval of 0.1 s (10 Hz) were repeatedly applied with intervals of 0.1 to 20 s between trains, the effect of GMA to increase ACh release depended on the length of the train interval; the longer the resting period between trains the higher was the output of ACh. This fact indicates that the release of ACh increased primarily during the resting periods following single stimuli or trains. The effect of GMA on ACh release proved to be highly temperature-dependent: in the presence of GMA Q10 increasing from 3.25 to 4.92. A high Ca concentration, removal of Mg or lowering of the Na concentration abolished the effect of GMA to enhance ACh release.     

The pedunculopontine nucleus projection to the parafascicular nucleus of the thalamus: an electrophysiological investigation in the rat

Summary. Extracellular electrophysiological recordings of neurons of the parafascicular nucleus of the thalamus were done in normal rats and in rats bearing lesions of either the cerebellar nuclei or the entopeduncular nucleus to investigate the functional control of the pedunculopontine nucleus on the parafascicular nucleus. A total of 97 neurons were recorded in the parafascicular nucleus in intact rats, 83 in rats bearing a chronic electrolytic lesion of the ipsilateral deep cerebellar nuclei, and 69 in rats bearing an ibotenate lesion of the ipsilateral entopeduncular nucleus. Lesions of the cerebellar nuclei or the entopeduncular nucleus were made to evaluate the participation of cerebellothalamic fibers or of polysynaptic basal ganglia circuits in the responses recorded in parafascicular neurons following electrical microstimulation of the ipsilateral pedunculopontine nucleus. Two types of excitation and one type of inhibition were the main responses observed in neurons of the parafascicular nucleus following stimulation of the pedunculopontine nucleus. The first type of excitation, observed in 49.5% of neurons recorded in normal rats, had an onset of 1.8 ± 0.6 ms, lasted 9.2 ± 0.8 ms and was able to follow high frequency stimulation over 300 Hz. The second type of excitation, observed in a smaller percentage of neurons recorded (3.1%), was a long-latency (8.3 ± 0.7 ms) activation lasting 19.0 ± 4.5 ms. It did not follow stimulation frequencies higher than 50–100 Hz. The inhibitory response was observed in 17.5% of the neurons recorded. The latency of this inhibition was 4.5 ± 1.8 ms and the duration 41.9 ± 6.8 ms. In rats bearing a lesion of the deep cerebellar nuclei or of the entopeduncular nucleus, the short-latency activation was still present in 24.1% and 31.9% of neurons recorded, respectively. However, the occurrence of the long-latency excitation rats bearing lesions of either the cerebellum or the entopeduncular nucleus increased to 12.1% and to 17.4%, respectively, while the occurrence of the inhibition rose to 22.9% and to 28.9%. These results show that an excitatory influence on the parafascicular nucleus is exerted by the pedunculopontine nucleus irrespectively of the presence of cerebellofugal fibers. This influence appears to be also independent from the integrity of basal ganglia circuits having a relay at the level of the entopeduncular nucleus. However, the variety of responses recorded suggests that the influences of the pedunculopontine nucleus on the parafascicular nucleus are by far more complex than those exerted on its basal ganglia targets such as the substantia nigra. The results are discussed according to a model of functioning of pedunculopontine fibers directed to thalamic and basal ganglia nuclei. Received December 16, 2002; accepted February 5, 2003 Published online April 22, 2003 Acknowledgements The present study has been supported by grants from Telethon (E0930), Ministero della Salute and Regione Lazio (Progetto Alzheimer), and Progetti d'Ateneo 2001–2002. A. C. and T. F. equally contributed to this work. Authors' address: Dr. T. Florio, Department of Biomedical Technology, University of L'Aquila, Via Vetoio Coppito 2, I-67100 L'Aquila, Italy, e-mail: Florio@univaq.it     

The effect of chronic atypical antipsychotic drugs and haloperidol on amphetamine-induced dopamine release in vivo.

The effect of chronic administration of antipsychotic drugs (21 days in drinking water followed by 3 days drug washout) on the D-amphetamine (1.0 mg/kg, s.c.)-induced increase in dopamine (DA) release in the striatum and the nucleus accumbens of awake, freely-moving rats was investigated with microdialysis. Chronic administration of haloperidol, a typical antipsychotic, (0.5 mg/kg/day), decreased basal extracellular DA release in the striatum and the nucleus accumbens but did not affect D-amphetamine-induced DA release in either region. In marked contrast, chronic administration of three atypical antipsychotic drugs: amperozide (2 mg/kg/day), clozapine (10 mg/kg/day) and melperone (2 mg/kg/day) increased basal extracellular DA and enhanced D-amphetamine-induced DA release in the striatum. In the nucleus accumbens, basal extracellular DA was decreased by chronic amperozide, unchanged by chronic clozapine and increased by chronic melperone. Most significantly, D-amphetamine-induced DA release was inhibited by chronic amperozide or clozapine, but unaffected by chronic melperone in this region. These results suggest that atypical antipsychotic drugs can alter DA release in a region specific manner. In particular, attenuation of amphetamine-like stimulation of DA release with reduced basal DA release in the nucleus accumbens could contribute to the antipsychotic action of amperozide which has a very weak affinity for D2 DA receptors.     

The primate centromedian-parafascicular complex: anatomical organization with a note on neuromodulation

In addition to the cerebral cortex, the striatum receives excitatory input from the thalamus. The centromedian (centre median, CM) and parafascicular (Pf) nuclei are an important source of thalamostriatal projections. Anterograde tract-tracing indicates the CM-Pf complex provides dense afferents to the matrix compartment of the striatum. Whereas CM projects to the entire sensorimotor territory of the striatum, the Pf provides complementary input to the entire associative sector. The Pf also provides lighter input to the nucleus accumbens. Both CM and Pf provide light to moderately dense inputs to other components of the basal ganglia in a largely complementary manner, covering motor or associative-limbic territories of the subthalamic nucleus, globus pallidus and ventral midbrain. In turn, the CM and Pf receive mainly segregated input from parallel motor and associative-limbic circuits of the basal ganglia. The CM and Pf may therefore be considered important participants in parallel processing of motor and associative-limbic information in the basal ganglia. Connections of the CM and Pf with other thalamic nuclei suggest they also participate in integrative functions within the thalamus. In addition, inputs from the brainstem reticular core, reciprocal connections with the cerebral cortex and reticular thalamic nucleus suggest a role in state-dependant information processing. Consideration of the differential connections of the CM and Pf, and better understanding of their role in pathophysiology, may eventually lead to development of an important new target for relief of a variety of neurological and psychiatric disorders.     

Physiological release of striatal acetylcholine (in vivo): effect of somatostatin on dopaminergic-cholinergic interaction

The effects of somatostatin (SOM) on the release of acetylcholine (ACh) and dopamine (DA) from striatum of freely moving rats were studied by transversal microdialysis. Acetylcholine (ACh) and dopamine (DA) were detected by high performance liquid chromatography (HPLC) with electrochemical detection. Somatostatin (0.1, 0.5 and 1 microM), administered locally through the microdialysis probe to the striatum, was able to release dose-dependently ACh from the cholinergic neurons of the striatum. The increase in the extracellular levels of ACh produced by 1 microM SOM in the striatum reached a maximum of 200%. ACh-releasing effect of SOM was completely inhibited by tetrodotoxin indicating that neuronal firing is involved in its effect. Local infusion of sulpiride, 10 microM, D(2) receptor antagonist, potentiated (about 100%) the SOM (1 microM)-induced release of ACh. SOM, 1 microM, was more effective in enhancing the release of ACh in the striatum (two-fold increase) after degeneration of the nigrostriatal DA pathway with 6-hydroxydopamine (6-OHDA) (250 microg/animal, i.c.v.). The D(2) receptor agonists bromcriptine, 10 microM, or apomorphine, 10 microM, completely antagonize SOM-induced release. SOM, 1 microM, enhanced the release of DA (about 400%). These findings indicate that SOM is capable of releasing both ACh and DA in the striatum, however, its effect on ACh release is partially masked unless the D(2) receptor-mediated tonic inhibitory effect of released DA from the nigro-striatal pathway is attenuated.     

Non-NMDA glutamatergic receptors modulate acetylcholine release in the rat subfornical organ area

The present study was designed to examine whether glutamatergic receptor mechanisms modulate the release of acetylcholine (ACh) in the region of the subfornical organ (SFO) using intracerebral microdialysis methods in freely moving rats. Perfusion of either non-N-methyl-d-aspartate (NMDA) agonist quisqualic acid (QA, 50 microM) or kainic acid (KA, 50 microM) through the microdialysis probe significantly enhanced the ACh release in the SFO area. Local perfusion of the non-NMDA receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX, 10 and 50 microM) did not change the basal release of ACh. CNQX (10 microM) administered together with either QA (50 microM) or KA (50 microM) in the SFO area antagonized the stimulant effect of the agonists on the ACh release. In urethane-anesthetized rats, repetitive electrical stimulation (500 microA, 10 Hz) of the medial septum (MS) significantly increased dialysate ACh concentrations in the region of the SFO. The increase in the ACh release elicited by the MS stimulation was significantly potentiated by perfusion of QA (50 microM), and the QA-induced potentiation was prevented by CNQX (10 microM) treated together with QA. These results show that the glutamatergic synaptic inputs enhance the ACh release in the SFO area through non-NMDA receptors. The data further suggest that the septal cholinergic inputs to the SFO area are potentiated by non-NMDA receptor mechanisms.     

A rat brain slice preparation for characterizing both thalamostriatal and corticostriatal afferents

The striatum, the primary input nucleus of the basal ganglia, is crucially involved in motor and cognitive function and receives significant glutamate input from the cortex and thalamus. Increasing evidence suggests fundamental differences between these afferents, yet direct comparisons have been lacking. We describe a slice preparation that allows for direct comparison of the pharmacology and biophysics of these two pathways. Visualization of slices from animals previously injected with BDA into the parafascicular nucleus revealed the presence of axons of thalamic origin in the slice. These axons were especially well-preserved after traversing the reticular nucleus, the location chosen for stimulation of thalamostriatal afferents. Initial characterization of the two pathways revealed both non-NMDA and NMDA receptor-mediated currents at synapses from both afferents and convergence of the afferents in 51% of striatal efferent neurons. Annihilation of action potentials was not observed in collision experiments, nor was current spread from the site of stimulation to striatum found. Differences in short-term plasticity suggest that the probability of release differs for the two inputs. The present work thus provides a novel rat brain slice preparation in which the effects of selective stimulation of cortical versus thalamic afferents to striatum can be studied in the same preparation.     

In vivo release of serotonin in cat dorsal vagal complex and cervical ventral horn induced by electrical stimulation of the medullary raphe nuclei.

Extracellular levels of serotonin (5-hydroxytryptamine; 5-HT) were monitored by microdialysis in the dorsal vagal complex (DVC) and the ventral horn of the spinal cord at the level of the phrenic motor nucleus in decerebrated cats. A selective serotonin uptake inhibitor, alaproclate (10(-4) M) was included in the dialysis probe perfusion fluid to increase basal and stimulated levels of 5-HT. Electrical stimulation (30 Hz, 10 V, 0.5 ms) in the nucleus raphe obscurus, containing neurons projecting to the DVC and to the ventral horn, induced a 2-3-fold increase of the 5-HT release in both these regions. After termination of the stimulation, the release gradually decreased during the following 60 min. Substance P, which coexists with 5-HT in descending neurons, did not significantly affect the 5-HT release when it was added (100 microM) to the probe perfusion fluid. The present findings are in accordance with the hypothesis that prolonged release of 5-HT is responsible for the previously demonstrated long-lasting facilitation of phrenic activity following raphe obscurus stimulation.     

Activation of the Subthalamic Nucleus and Pedunculopontine Tegmentum: Does it Affect Dopamine Levels in the Substantia Nigra,Nucleus Accumbens and Striatum?

Parkinson's disease is a neurodegenerative disorder, of which the most prominent morphological feature is the progressive loss of dopaminergic nigrostriatal neurons. Increased glutamatergic transmission in the basal ganglia has been implicated in the pathophysiology of Parkinson's disease (PD). This study investigated whether death of substantia nigra (SN) dopaminergic neurons could be caused by the hyperactivity of afferent pathways resulting in the release of a toxic dose of excitatory amino acids in the SN. Twice-daily unilateral stimulation of the subthalamic nucleus (STN) for 21 days, using two different pulse frequencies and current strengths, significantly increased amphetamine-induced rotation, whereas sham stimulated rats showed significantly reduced rotation. Striatal and SN dopamine (DA) levels were unaffected when compared to naı̈ve and sham stimulated rats. However, levels of the DA metabolite, homovanillic acid (HVA), were significantly higher in the ipsilateral anterior striata of rats that had been stimulated at high frequency (100 Hz) and low current (100 μA) as compared to sham treated animals. Stimulation of the pedunculopontine tegmentum (PPT), using a single kainic acid injection, did not affect DA concentration in the ipsilateral striatum and nucleus accumbens when compared to sham-treated rats. DA levels in the contralateral striatum and nucleus accumbens of lesioned rats were significantly higher than ipsilateral levels. DOPAC/DA ratios were lower in the contralateral striatum and nucleus accumbens, suggesting decreased DA turnover. Glutamic acid decarboxylase activity was significantly higher in the ipsilateral than the contralateral SN. The physical manifestations of PD require a large reduction in caudate and putamen DA levels and no such depletion was measured in this study. These results, therefore, do not support the hypothesis that Parkinson's disease may result from an overstimulation of substantia nigral DAneurons by glutamate afferents originating from the STN or PPT.     

Nitric oxide (NO) increases acetylcholine release from and inhibits smooth muscle contraction of guinea-pig gastric fundus

Experiments were carried out to investigate the interaction between nitric oxide (NO) and cholinergic neurotransmission in smooth muscle strips of guinea-pig gastric fundus. Electrical field stimulation (2 Hz, 1 ms, 360 shocks) evoked atropine-sensitive contractions. Dimethylphenylpiperazinium (DMPP) (100 microM), a nicotinic receptor agonist, reversed the stimulation-evoked contraction and resulted in relaxation. Nomega-nitro-L-arginine (L-NNA) (100 microM), an NO synthase inhibitor, significantly increased the amplitude of stimulation-evoked contraction and abolished the effect of DMPP. Electrical stimulation increased the release of [3H]acetylcholine ([3H]ACh) from the tissue strips above the basal levels. Neither L-NNA (100 microM) nor DMPP (100 microM) alone influenced the basal release of [3H]ACh. Nomega-nitro-L-arginine (100 microM) decreased the electrical stimulation-evoked release of [3H]ACh. Dimethylphenylpiperazinium increased the stimulation-evoked release of [3H]ACh but had no effect in the presence of L-NNA. It is suggested that in guinea-pig gastric fundus, endogenous NO released in response to field stimulation has an opposite effect at the pre- and postsynaptic sites: it increases the release of ACh from cholinergic nerve terminals but reduces smooth muscle responses to ACh.     

A within-subjects microdialysis/behavioural study of the role of striatal acetylcholine in D1-dependent turning.

In rats lesioned with 6-hydroxydopamine (6-OHDA) the effect of the noncompetitive N-methyl D-aspartate (NMDA) receptor antagonist, MK-801, the dopamine (DA) D2 receptor agonist quinpirole and the A2A adenosine antagonist SCH 58261 was studied on acetylcholine (ACh) release in the lesioned striatum and contralateral turning behaviour stimulated by the administration of the DA D1 receptor agonist CY 208-243. Administration of CY 208-243 (75, 100 and 200 microg/kg) to 6-OHDA-lesioned rats dose-dependently stimulated ACh release and induced contralateral turning. MK-801 (50 and 100 microg/kg) reduced basal ACh release (max 22%) and did not elicit any turning. MK-801 (50 and 100 microg/kg) potentiated the contralateral turning, but failed to modify the stimulation of ACh release elicited by 100 and 200 microg/kg of CY 208-243. MK-801 (100 microg/kg) prevented the increase in striatal ACh release evoked by the lower dose of CY 208-243 (75 microg/kg) but contralateral turning was not observed. The D2 receptor agonist quinpirole (30 and 60 microg/kg) elicited low-intensity contralateral turning and decreased basal ACh release. Quinpirole potentiated the D1-mediated contralateral turning behaviour elicited by CY 208-243 (100 microg/kg), but failed to affect the increase in ACh release elicited by the D1 agonist. The adenosine A2A receptor antagonist SCH 58261 (1 microg/kg i.v.) failed per se to elicit contralateral turning behaviour. SCH 58261 potentiated the contraversive turning induced by CY 208-243 but failed to affect the increase of ACh release. The results of the present study indicate that blockade of NMDA receptors by MK-801. stimulation of DA D2 receptors by quinpirole and blockade of adenosine A2A receptors by SCH 58261 potentiate the D1-mediated contralateral turning behaviour in DA denervated rats without affecting the action of the D1 agonist on ACh release. These observations do not support the hypothesis that the potentiation of D1-dependent contralateral turning by MK-801, quinpirole or SCH 58261 is mediated by changes in D1-stimulated release of ACh in the striatum.     

Response of nucleus accumbens neurons to amygdala stimulation and its modification by dopamine

Extracellular single unit recordings were obtained from the nucleus accumbens of urethane anesthetized rats. It was found that electrical stimulation of the basal lateral and basal medial nuclei of the amygdala produced strong excitatory responses in neurons of the nucleus accumbens, in particular the medial region. Latencies of activation were relatively short with a mean of 10.7 ms.Dopamine applied iontophoretically had a marked attenuating effect on the excitatory response of nucleus accumbens neurons to amygdala stimulation. The spontaneous activity of all neurons recorded from the nucleus accumbens was also suppressed by dopamine, but the excitatory response was more sensitive to dopamine inhibition than the spontaneous activity.Neurons in the nucleus accumbens showed a variety of responses to single-pulse electrical stimulation of the ventral tegmental area (VTA). Some units in the nucleus accumbens received convergent inputs from both the amygdala and the VTA. Stimulation of the VTA also attenuated the response of nucleus accumbens neurons to excitatory inputs from the amygdala. A train of 10 pulses (0.15 ms, 200–600 αA) at 10 Hz delivered to the VTA at 100 ms before stimulation of the amygdala caused attenuation of the original excitatory response. The attenuating effect could be observed irrespective of whether individual single-pulse stimulation of the VTA elicited a response in that particular accumbens neuron or not. 6-Hydroxydopamine injected into the VTA 2 days prior to the recording experiment, or haloperidol injected intraperitoneally 1 h before the recording session, abolished this attenuating effect. However, responses to single-pulse stimulations of the VTA were not abolished. The results suggest that the attenuation of the excitatory response to amygdala stimulation was due to the release of dopamine from mesolimbic dopaminergic neurons. Responses to single-pulse stimulations of the VTA were probably due to activation of non-dopaminergic neurons projecting from the same area.It is suggested as a working hypothesis that this inhibitory effect of dopamine may be an important function of the mesolimbic dopamine pathway in modulating the extent to which limbic structures can exert an influence on the motor system through the accumbens.     

Differences in the regulation of acetylcholine release upon D2 dopamine and N-methyl-D-aspartate receptor activation between the striatal complex of reptiles and the neostriatum of rats.

Activation of the N-methyl-D-aspartate (NMDA) receptor increases and that of the D2 dopamine (DA) receptor inhibits the release of acetylcholine (ACh) from mammalian neostriatal tissue. Similar effects have been described in the ventral striatum of the rat, however, in the caudomedial part of the nucleus accumbens, D2 receptor activation does not inhibit the release of ACh. Likewise, the NMDA-induced stimulation of the release of ACh in this part of the striatum is much smaller. In the present study we demonstrated that in the striatal complex or striatum of reptiles D2 receptor activation did not result in an inhibition of the release of ACh, whereas the release of DA could be inhibited to a significant extent. These findings indicate that, although D2 receptors are present in the striatum of reptiles, these receptors do not regulate the release of ACh in this brain structure. We observed in the striatum of reptiles a potassium induced and calcium-dependent release of [3H]D-aspartate indicating a neurotransmitter role for aspartate or glutamate (GLU). However GLU and NMDA have only a marginal effect on the release of ACh in the striatum of the reptiles as compared to the effects in the neostriatum of the rat. It is concluded that with respect to the effects of D2 and NMDA receptor activation on the release of ACh, the striatum of reptiles bears most similarity to the caudomedial part of the nucleus accumbens.