Safinamide Modulates Striatal Glutamatergic Signaling in a Rat Model of Levodopa-Induced Dyskinesia.

Safinamide (Xadago) is a novel dual-mechanism drug that has been approved in the European Union and United States as add-on treatment to levodopa in Parkinson's disease therapy. In addition to its selective and reversible monoamine oxidase B inhibition, safinamide through use-dependent sodium channel blockade reduces overactive glutamatergic transmission in basal ganglia, which is believed to contribute to motor symptoms and complications including levodopa-induced dyskinesia (LID). The present study investigated the effects of safinamide on the development of LID in 6-hydroxydopamine (6-OHDA)-lesioned rats, evaluating behavioral, molecular, and neurochemical parameters associated with LID appearance. 6-OHDA-lesioned rats were treated with saline, levodopa (6 mg/kg), or levodopa plus safinamide (15 mg/kg) for 21 days. Abnormal involuntary movements, motor performance, molecular composition of the striatal glutamatergic synapse, glutamate, and GABA release were analyzed. In the striatum, safinamide prevented the rearrangement of the subunit composition of N-methyl-d-aspartate receptors and the levodopa-induced increase of glutamate release associated with dyskinesia without affecting the levodopa-stimulated motor performance and dyskinesia. Overall, these findings suggest that the striatal glutamate-modulating component of safinamide's activity may contribute to its clinical effects, where its long-term use as levodopa add-on therapy significantly improves motor function and "on" time without troublesome dyskinesia.

Development of a new toolbox for mouse PET-CT brain image analysis fully based on CT images and validation in a PD mouse model.

Automatic analysis toolboxes are popular in brain image analysis, both in clinical and in preclinical practices. In this regard, we proposed a new toolbox for mouse PET-CT brain image analysis including a new Statistical Parametric Mapping-based template and a pipeline for image registration of PET-CT images based on CT images. The new templates is compatible with the common coordinate framework (CCFv3) of the Allen Reference Atlas (ARA) while the CT based registration step allows to facilitate the analysis of mouse PET-CT brain images. From the ARA template, we identified 27 volumes of interest that are relevant for in vivo imaging studies and provided binary atlas to describe them. We acquired 20 C57BL/6 mice with [18F]FDG PET-CT, and 12 of them underwent 3D T2-weighted high-resolution MR scans. All images were elastically registered to the ARA atlas and then averaged. High-resolution MR images were used to validate a CT-based registration pipeline. The resulting method was applied to a mouse model of Parkinson's disease subjected to a test-retest study (n = 6) with the TSPO-specific radioligand [18F]VC701. The identification of regions of microglia/macrophage activation was performed in comparison to the Ma and Mirrione template. The new toolbox identified 11 (6 after false discovery rate adjustment, FDR) brain sub-areas of significant [18F]VC701 uptake increase versus the 4 (3 after FDR) macro-regions identified by the Ma and Mirrione template. Moreover, these 11 areas are functionally connected as found by applying the Mouse Connectivity tool of ARA. In conclusion, we developed a mouse brain atlas tool optimized for PET-CT imaging analysis that does not require MR. This tool conforms to the CCFv3 of ARA and could be applied to the analysis of mouse brain disease models.

Identification of distinct pathological signatures induced by patient-derived α-synuclein structures in nonhuman primates.

Dopaminergic neuronal cell death, associated with intracellular α-synuclein (α-syn)-rich protein aggregates [termed "Lewy bodies" (LBs)], is a well-established characteristic of Parkinson's disease (PD). Much evidence, accumulated from multiple experimental models, has suggested that α-syn plays a role in PD pathogenesis, not only as a trigger of pathology but also as a mediator of disease progression through pathological spreading. Here, we have used a machine learning-based approach to identify unique signatures of neurodegeneration in monkeys induced by distinct α-syn pathogenic structures derived from patients with PD. Unexpectedly, our results show that, in nonhuman primates, a small amount of singular α-syn aggregates is as toxic as larger amyloid fibrils present in the LBs, thus reinforcing the need for preclinical research in this species. Furthermore, our results provide evidence supporting the true multifactorial nature of PD, as multiple causes can induce a similar outcome regarding dopaminergic neurodegeneration.

Nocistatin inhibits 5-hydroxytryptamine release in the mouse neocortex via presynaptic Gi/o protein linked pathways.

BACKGROUND AND PURPOSE: Nocistatin (NST) is a neuropeptide generated from cleavage of the nociceptin/orphanin FQ (N/OFQ) precursor. Evidence has been presented that NST acts as a functional antagonist of N/OFQ, although NST receptor and transduction pathways have not yet been identified. We previously showed that N/OFQ inhibited [(3)H]5-hydroxytryptamine ([(3)H]5-HT) release from mouse cortical synaptosomes via activation of NOP receptors. We now investigate whether NST regulates [(3)H]5-HT release in the same preparation. EXPERIMENTAL APPROACH: Mouse and rat cerebrocortical synaptosomes in superfusion, preloaded with [(3)H]5-HT and stimulated with 1 min pulses of 10 mM KCl, were used. KEY RESULTS: Bovine NST (b-NST) inhibited the K(+)-induced [(3)H]5-HT release, displaying similar efficacy but lower potency than N/OFQ. b-NST action underwent concentration-dependent and time-dependent desensitization, and was not prevented either by the NOP receptor antagonist [Nphe(1) Arg(14),Lys(15)]N/OFQ(1-13)-NH(2) (UFP-101) or by the non-selective opioid receptor antagonist, naloxone. Contrary to N/OFQ, b-NST reduced [(3)H]5-HT release from synaptosomes obtained from NOP receptor knockout mice. However, both N/OFQ and NST were ineffective in synaptosomes pre-treated with the G(i/o) protein inhibitor, Pertussis toxin. NST-N/OFQ interactions were also investigated. Co-application of maximal concentrations of both peptides did not result in additive effects, whereas pre-application of maximal b-NST concentrations partially attenuated N/OFQ inhibition. CONCLUSIONS AND IMPLICATIONS: We conclude that b-NST inhibits [(3)H]5-HT release via activation of G(i/o) protein linked pathways, not involving classical opioid receptors and the NOP receptor. The present data strengthen the view that b-NST is, per se, a biologically active peptide endowed with agonist activity.

Synthesis and structure-activity relationships of deltorphin analogues.

In order to study the structure-activity relationships of natural opioid deltorphins (H-Tyr-D-Met-Phe-His-Leu-Met-Asp-NH2 and H-Tyr-D-Ala-Phe-Asp [or Glu]-Val-Val-Gly-NH2), 15 analogues were synthesized by the solution method. Their activities were determined in binding studies based on displacement of mu- and delta-receptor selective radiolabels from rat brain membranes and in two bioassays, using guinea pig ileum and mouse vas deferens. The obtained data indicate that the high delta-selectivity of deltorphins can be due to the constitution/conformation of the C-terminal part and, at least in part, to preselection by charge.

Nociceptin/orphanin FQ receptors modulate glutamate extracellular levels in the substantia nigra pars reticulata. A microdialysis study in the awake freely moving rat.

Intracerebral microdialysis was employed in awake freely moving rats to investigate the effects of nociceptin/orphanin FQ receptor ligands on glutamate extracellular levels in the substantia nigra pars reticulata. Nociceptin/orphanin FQ, ineffective at 0.1 microM, induced a prolonged stimulation of nigral glutamate levels at 1 and 10 microM (mean effect of 137+/-9 and 167+/-13%, respectively, of basal values). These effects were prevented by the novel nociceptin/orphanin FQ receptor antagonist [Nphe(1)]nociceptin/orphanin FQ(1-13)NH(2) (100 and 300 microM, respectively) but not by the non-selective opioid receptor antagonist naloxone (10 microM). [Nphe(1)]nociceptin/orphanin FQ(1-13)NH(2) (100 microM) inhibited by itself glutamate outflow (maximal reduction to 71+/-4%) while naloxone was ineffective. The nociceptin/orphanin FQ receptor ligand [Phe(1)psi(CH(2)-NH)Gly(2)]nociceptin/orphanin FQ(1-13)NH(2) also facilitated glutamate outflow at 10 microM (mean effect of 145+/-10%). Intranigral perfusion with tetrodotoxin (1 microM) or with the dopamine D(2) receptor antagonist raclopride (1 microM), failed to affect basal glutamate output and prevented the facilitatory effect of nociceptin/orphanin FQ (10 microM). However, perfusion with the GABA(A) receptor antagonist bicuculline (10 microM) increased local glutamate extracellular levels by itself and attenuated the effect of the peptide. Our data suggest that nociceptin/orphanin FQ increases glutamate extracellular levels in the substantia nigra pars reticulata via activation of nociceptin/orphanin FQ receptors located on non-glutamatergic, possibly dopaminergic and GABAergic, neuronal elements.

Functional neuroanatomy of the nigrostriatal and striatonigral pathways as studied with dual probe microdialysis in the awake rat--I. Effects of perfusion with tetrodotoxin and low-calcium medium.

In the present study we employed the dual probe approach to investigate functional interactions between the nigrostriatal dopaminergic and striatonigral GABAergic pathways in the awake, freely moving rat and their role in motor function. One microdialysis probe of concentric design was implanted in the substantia nigra pars reticulata and another in the ipsilateral dorsolateral striatum. Perfusion with a low-Ca2+ (0.1 mM) medium and with the voltage-dependent Na(+)-channel blocker tetrodotoxin (10 microM) was alternatively performed in both brain regions and the dialysate dopamine, glutamate and GABA levels were simultaneously measured in the dorsolateral striatum, whereas GABA levels alone were monitored in the substantia nigra. Perfusion with a low-Ca2+ medium in the substantia nigra pars reticulata did not affect local GABA levels, but transiently increased striatal dopamine release (+40%) without modifying striatal glutamate and GABA levels. Conversely, intranigral perfusion with tetrodotoxin transiently increased local GABA levels (+40%), while it decreased striatal dopamine (-60%) and increased glutamate (+70%) and GABA (+50%) levels. Perfusion with a low-Ca2+ medium in the dorsolateral striatum reversibly decreased local dopamine (-70%), glutamate (-20%) and GABA (-20%) levels, while local perfusion with tetrodotoxin decreased dopamine (-70%), increased glutamate (+30%) but did not affect dialysate GABA levels in this brain area. Neither of these intrastriatal treatments significantly affected GABA levels in the substantia nigra. Intranigral but not intrastriatal perfusion with tetrodotoxin was also associated with an increase in spontaneous locomotor activity as expressed by contralateral turning. Intranigral and intrastriatal perfusion with low-Ca2+ medium did not influence locomotor activity. On the basis of these neurochemical and behavioural findings, we propose a new dynamic model for the study of motor behaviour as mediated by basal ganglia circuitry.

Functional neuroanatomy of the nigrostriatal and striatonigral pathways as studied with dual probe microdialysis in the awake rat--II. Evidence for striatal N-methyl-D-aspartate receptor regulation of striatonigral GABAergic transmission and motor function.

In the present study we used the dual probe approach to investigate striatal N-methyl-D-aspartate receptor regulation of GABA release from the substantia nigra pars reticulata of the awake, freely moving rat. One microdialysis probe of concentric design was implanted in the dorsolateral striatum and another in the ipsilateral substantia nigra pars reticulata. Perfusion with N-methyl-D-aspartate (100 microM) in the dorsolateral striatum decreased local dopamine release (-25%) and increased both glutamate (+40%) and GABA (+35%) release. Moreover, perfusion with N-methyl-D-aspartate (100 microM) in the dorsolateral striatum increased GABA release (+20%) in the substantia nigra pars reticulata. Perfusion with the lower (10 microM) N-methyl-D-aspartate concentration in the dorsolateral striatum did not affect striatal dopamine, glutamate and GABA release or nigral GABA release. Intrastriatal perfusion with the N-methyl-D-aspartate receptor antagonist dizocilpine maleate (10 microM), at a dose which by itself did not affect basal striatal or nigral neurotransmitter levels, prevented the effects of striatal perfusion with N-methyl-D-aspartate on both striatal and nigral neurotransmitter release. Intrastriatal dizocilpine maleate was also perfused concurrently with intranigral tetrodotoxin (10 microM) (see accompanying paper). Intrastriatal perfusion with dizocilpine maleate prevented the tetrodotoxin-induced rise in both striatal and nigral GABA levels and profoundly reduced the tetrodotoxin-induced contralateral turning. In addition, intrastriatal dizocilpine maleate delayed the increase in striatal glutamate release evoked by intranigral tetrodotoxin without affecting the associated decrease in striatal dopamine release. The present study demonstrates that N-methyl-D-aspartate receptors in the dorsolateral striatum regulate GABA release in the substantia nigra pars reticulata of the awake rat and provides evidence that this regulation plays a key role in motor function.

5-Hydroxytryptamine-mediated effects of nicotine on endogenous GABA efflux from guinea-pig cortical slices.

1. The effect of nicotine on endogenous basal GABA outflow was studied in guinea-pig cerebral cortex slices. 2. Nicotine 1.86-18.6 mumol l-1 significantly decreased the basal, tetrodotoxin-sensitive GABA efflux, whereas at higher concentrations (186-620 mumol l-1) nicotine increased it. The inhibition was prevented by mecamylamine while the facilitation was blocked by mecamylamine, (+)-tubocurarine and tetrodotoxin. 3. The effect of nicotine was due to an indirect 5-hydroxytryptaminergic action. In fact, MDL 72222 (1 mumol l-1) completely prevented the alkaloid inhibition and methysergide (1 mumol l-1) reversed the facilitation into inhibition; concomitant treatment with methysergide and MDL 72222 antagonized the effect of nicotine at 186 mumol l-1 4. Lower concentrations of 5-HT (3-10 mumol l-1) decreased, whereas higher concentrations (30-100 mumol l-1) increased, spontaneous GABA outflow. The inhibition of GABA efflux was prevented by MDL 72222 whereas the facilitation was reversed by methysergide (1 mumol l-1) into inhibition, and prevented by MDL 72222 1 mumol l-11. 5. These results suggest that, by activating nicotinic receptors present on 5-hydroxytryptaminergic terminals, nicotine releases 5-HT which, in turn, inhibits or increases the secretory activity of cortical GABA interneurones via 5-HT3 and methysergide-sensitive receptors, respectively.

Modulation of 5-hydroxytryptamine efflux from rat cortical synaptosomes by opioids and nociceptin.

The modulation of [(3)H]-5-hydroxytryptamine ([(3)H]-5-HT) efflux from superfused rat cortical synaptosomes by delta, kappa, mu and ORL(1) opioid receptor agonists and antagonists was studied. Spontaneous [(3)H]-5-HT efflux was reduced (20% inhibition) by either 0.5 microM tetrodotoxin or Ca(2+)-omission. Ten mM K(+)-evoked [(3)H]-5-HT overflow was largely Ca(2+)-dependent (90%) and tetrodotoxin-sensitive (50%). The delta receptor agonist, deltorphin-I, failed to modulate the K(+)-evoked neurotransmitter efflux up to 0.3 microM. The kappa and the mu receptor agonists, U-50,488 and endomorphin-1, inhibited K(+)-evoked [(3)H]-5-HT overflow (EC(50)=112 and 7 nM, respectively; E(max)=28 and 29% inhibition, respectively) in a norBinaltorphimine- (0.3 microM) and naloxone- (1 microM) sensitive manner, respectively. None of these agonists significantly affected spontaneous [(3)H]-5-HT efflux. The ORL(1) receptor agonist nociceptin inhibited both spontaneous (EC(50)=67 nM) and K(+)-evoked (EC(50)=13 nM; E(max)=52% inhibition) [(3)H]-5-HT efflux. The effect of NC was insensitive to naloxone (up to 10 microM), but was antagonized by [Nphe(1)]nociceptin(1-13)NH(2) (a novel selective ORL(1) receptor antagonist; pA(2)=6.7) and by naloxone benzoylhydrazone (pA(2)=6.3). The ORL(1) ligand [Phe(1)psi(CH(2)-NH)Gly(2)]nociceptin(1-13)NH(2) also inhibited K(+) stimulated [(3)H]-5-HT overflow (EC(50)=64 nM; E(max)=31% inhibition), but its effect was partially antagonized by 10 microM naloxone. It is concluded that the ORL(1) receptor is the most important presynaptic modulator of neocortical 5-HT release within the opioid receptor family. This suggests that the ORL(1)/nociceptin system may have a powerful role in the control of cerebral 5-HT-mediated biological functions.

Effect of acute and subchronic nicotine treatment on cortical efflux of [3H]-D-aspartate and endogenous GABA in freely moving guinea-pigs.

1. The [3H]-D-aspartate preloading of the parietal cortex of freely moving guinea-pigs equipped with epidural cups makes it possible to investigate drug effects on the efflux of this radiolabel, assumed as a marker of the glutamatergic structures underlying the cup. In the same model, the efflux of [3H]-gamma-aminobutyric acid ([3H]-GABA) and endogenous GABA can be measured. 2. Nicotine, 0.9-3.6 mg kg-1, s.c., or 3-5 micrograms, i.c.v., increased the efflux of [3H]-D-aspartate but reduced that of GABA. 3. These effects were mediated through mecamylamine-sensitive receptors but the ganglionic blocking agent was devoid of any primary activity. 4. The inhibition of GABA efflux induced by nicotine 3.6 mg kg-1, s.c., was abolished by methysergide 2 mg kg-1, i.p. and was reduced by naloxone 3 mg kg-1, i.p. pretreatment, suggesting the involvement of tryptaminergic and opioid systems. In contrast, muscarinic and catecholamine antagonists were ineffective. 5. Chronic treatment with nicotine (3.6 mg kg-1, twice daily for 16 days) reduced the facilitatory effect of [3H]-D-aspartate and abolished the inhibition of endogenous GABA efflux. 6. A slight increase in the number of nicotinic binding sites (by use of [3H]-nicotine as ligand) was found in the neocortex of chronically treated guinea-pigs. 7. The higher degree of tolerance to chronic nicotine treatment shown by GABA as compared with [3H]-D-aspartate efflux suggests that adaptative changes of the inhibitory neuronal pools prevail. This may contribute to the reinforcing and addictive properties of nicotine.

AMPA receptor activation regulates the glutamate metabotropic receptor stimulated phosphatidylinositol turnover in human cerebral cortex slices.

The effect of excitatory amino acids (EAA) on phosphatidylinositol (PI) turnover in human cerebral cortical slices was investigated. Trans-1-aminocyclopentane-1,3-dicarboxylic acid (ACPD) increased inositol phosphate (IP) formation in the 1-1000 microM range. Quisqualic acid (QA) was maximally effective at 10-100 microM, showing an inverse correlation between concentration and effect in the 100-1000 microM range. The glutamate metabotropic receptor antagonist 2-amino-3-phosphonopropionic acid (AP3), the ionotropic non-NMDA receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) and the NMDA channel blocker dizolcipine (MK-801) failed to prevent the PI response to ACPD (1000 microM). However, CNQX (100 microM) modified the concentration-response curve of QA reducing the effect of QA 10 microM by approx. 50% and enhancing that of QA 1000 microM by 2-fold. In addition, CNQX (100 microM) together with MK-801 (100 microM) unmasked the ability of L-glutamate (L-GLU) 3000 microM to stimulate PI turnover. The effect of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) on the EAA-induced PI turnover was also studied. AMPA (0.1-1 microM) potentiated the response to submaximal (30 microM) ACPD and (1 microM) QA concentrations. However, higher AMPA concentrations (10 microM) failed to synergize with ACPD 30 microM and, in addition, inhibited the PI turnover maximally stimulated by QA 10 microM. These results further support the presence of the glutamate metabotropic receptor in the human neocortex. In addition, they show the occurrence of a concentration-related dual interaction between AMPA and glutamate metabotropic receptor activation in the IP formation in this brain area.

Reciprocal dopamine-glutamate modulation of release in the basal ganglia.

Dopaminergic and glutamatergic transmissions have long been known to interact at multiple levels in the basal ganglia to modulate motor and cognitive functions. One important aspect of their interactions is represented by the reciprocal modulation of release. This topic has been the object of interest since the late 70's, particularly in the striatum and in midbrain dopaminergic areas (substantia nigra and ventral tegmental area). Analysis of glutamate-dopamine interactions in the control of each other's release is complicated by the fact that both glutamate and dopamine act on multiple receptor subtypes which can exert different effects. Therefore, glutamatergic modulation of dopamine release has been reviewed by analyzing the effects of glutamatergic selective receptor agonists and antagonists in the striatum (both motor and limbic portions) and in midbrain dopaminergic areas, as revealed by in vitro (slices, cell cultures, synaptosomes) and in vivo (push-pull, microdialysis and voltammetry techniques) experimental approaches. The same approach has been followed for dopaminergic modulation of glutamate release. The facilitatory nature of glutamate modulating both presynaptic and dendritic dopamine release has clearly emerged from in vitro studies. However, evidence is presented that, at least in the striatum and in the nucleus accumbens of awake rats, glutamate-mediated inhibitory effects may also occur. In vitro and in vivo experiments in the striatum and midbrain dopaminergic areas mainly depict dopamine as an inhibitory modulator of glutamate release. However, in vivo studies reporting dopamine D1 receptor mediated facilitatory effects are also considered. Therefore, the general notion that glutamate and dopamine act oppositely to regulate each other's release, is only partly supported by the available data. Conversely, the nature of the interaction between the two neurotransmitters seems to vary depending on the experimental approach, the brain area considered and the subtype of receptor involved.

Excitatory amino acids (EAAs) stimulate phosphatidylinositol turnover in adult rat striatal slices: interaction between NMDA and EAA metabotropic receptors.

The effect of excitatory amino acids (EAAs) on phosphatidylinositol (PI) turnover in adult rat striatal slices was investigated. Quisqualic acid (QA), alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA), kainic acid (KA), ibotenic acid (IBO) and N-methyl-D-aspartic acid (NMDA) maximally increased inositol phosphate (IP) formation at 10 microM while trans-1-amino-cyclopentane-1,3-dicarboxylic acid (ACPD) was maximally effective at 100 microM. The NMDA channel blocker dizolcipine (MK-801) counteracted the effect of NMDA 10 microM and IBO 10 microM while it potentiated that of IBO 100 microM and IBO 1000 microM. Conversely, the non-NMDA receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) prevented the effect of AMPA and KA and reduced that of QA (all at 10 microM). Lowering extracellular Ca2+ concentrations ([Ca2+]0) differentially affected the PI response to EAAs. The ACPD 30 microM effect was unchanged at low [Ca2+]0 (but abolished when EGTA 2 mM was added), while that of ACPD 100 microM was halved in 0.1 mM and almost abolished in a nominally free Ca2+ medium. NMDA 10 microM and AMPA 10 microM were ineffective at low [Ca2+]0 while NMDA 100 microM, ineffective in a 1.2 mM Ca2+ medium, strongly stimulated IP formation in 0.1 mM Ca2+ but not in a nominally free Ca2+ medium. The effect of NMDA on EAA metabotropic receptor agonist stimulated PI turnover was also studied. NMDA 10 microM potentiated the effect of ACPD 30 microM. This positive cooperation persisted at low [Ca2+]0 but not in the presence of EGTA. Conversely, NMDA 100 microM prevented the effect of ACPD 100 microM. This negative interference was reversed when Ca2+ was omitted from the medium. This study shows that in the adult rat striatum both EAA metabotropic and ionotropic receptor activation increases IP formation. A positive and negative interaction between NMDA and metabotropic receptor activation was also found to regulate PI turnover. The role of [Ca2+]0 in subserving the PI response to EAAs was made evident.

Nociceptin/orphanin FQ and neurotransmitter release in the central nervous system.

In this article, the effect of nociceptin (orphanin FQ) on transmitter release in the central nervous system in vitro and in vivo is reviewed. Nociceptin inhibits the electrically or K(+)-evoked noradrenaline, dopamine, serotonin, and glutamate release in brain slices from guinea-pig, rat, and mouse. This effect is usually naloxone-resistant but antagonized by OP(4) receptor antagonists like [Phe(1)psi(CH(2)-NH)Gly(2)]-nociceptin(1-13)NH(2). In the rat in vivo, nociceptin diminishes acetylcholine release in the striatum, reduces dopamine release, and prevents the stimulatory effect of morphine on this transmitter in the nucleus accumbens and also elevates extracellular glutamate and gamma-aminobutyric acid levels in mesencephalic dopaminergic areas. The effect of nociceptin on the mesencephalic dopaminergic system might explain its actions on motor behavior.

Inhibitory effect of NMDA receptor activation on quisqualate-stimulated phosphatidylinositol turnover in the human cerebral cortex.

The effect of excitatory amino acids (EAA) on the phosphatidylinositol (PI) turnover in human cerebral cortical slices was investigated. Quisqualic acid (QA) and, to lesser extent, ibotenic acid (IBO) at 10(-5)-10(-3) M increased inositol phosphate (IP) accumulation. L-Glutamic acid (L-glu), kainic acid (KA), alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid and N-methyl-D-aspartic acid (NMDA) were ineffective. NMDA dose-dependently antagonized the QA facilitatory effect. Such inhibition was prevented by the NMDA receptor complex antagonists (+)-5-methyl-10,11-dihydro-5H-dibenzo[a, d]cyclohepten-5,10-imine (MK-801) and by 3[+/-)-2-carboxypiperazin-4-yl)propyl-1-phosphonic acid. The effect of IBO (but not that of QA) was greatly potentiated by MK-801. These data suggest that the EAA metabotropic receptor described in the rodent brain is also present in human cerebral cortex and is negatively modulated by the NMDA receptor.

Dopamine D1 and D2 receptor antagonism differentially modulates stimulation of striatal neurotransmitter levels by N-methyl-D-aspartic acid.

The effects of local perfusion with the dopamine D1 receptor antagonist SCH 23390 and the dopamine D2 receptor antagonist raclopride on basal and N-methyl-D-aspartate (NMDA) stimulated dopamine, gamma-aminobutyric acid (GABA) and glutamate levels in the dorsolateral striatum were monitored using in vivo microdialysis in the halothane anaesthetized rat. Local perfusion (90 min) with SCH 23390 and raclopride (1 and 10 microM) dose dependently increased basal striatal dopamine release whereas GABA and glutamate dialysate levels were unaffected. Local perfusion (10 min) with the 1 mM concentration of NMDA increased striatal dopamine, GABA and glutamate outflow. However, when perfused together with SCH 23390 (1 microM) NMDA inhibited glutamate efflux. Moreover, in the presence of SCH 23390 (10 microM) the NMDA induced rise in dopamine and GABA levels was partly prevented. On the other hand, raclopride 1 microM enhanced the NMDA stimulated GABA efflux while at 10 microM it partly counteracted the NMDA induced dopamine release. These data demonstrate that NMDA induced changes in striatal neurotransmitter outflow are effectively modified by dopamine D1 and D2 receptor blockade.

Glutamate antagonists prevent morphine withdrawal in mice and guinea pigs.

The effects of excitatory amino acid antagonists on increased cortical acetylcholine release and behavioral hyperactivity induced by naloxone in morphine tolerant guinea pigs and mice were studied. The results show that the N-methyl-D-aspartic acid (NMDA) antagonist MK-801 (0.1-1 mg/kg, i.p.) injected 30 min before naloxone (3 mg/kg, s.c.) dose-dependently prevented the neurochemical and behavioral signs of morphine withdrawal in guinea pigs and mice. The non-selective antagonist glutamic acid diethylester only at 100 mg/kg i.p. reduced the naloxone-induced increase of cortical acetylcholine release without affecting the behavioral changes. These findings indicate that the activation of excitatory amino acid receptors, mainly the NMDA receptors, plays a relevant role in the expression of opiate abstinence.

Identification and targeting policies for computer-controlled infusion pumps.

In this article a series of useful techniques is presented to improve the adaptation capabilities of computer-controlled infusion pumps: an identification algorithm for the time constant of the effect compartment, a smoothing technique for estimation based on noisy data, and an infusion policy to target any effect site concentration with no overshoot.

A reliable gait phase detection system.

A new highly reliable gait phase detection system, which can be used in gait analysis applications and to control the gait cycle of a neuroprosthesis for walking, is described. The system was designed to detect in real-time the following gait phases: stance, heel-off, swing, and heel-strike. The gait phase detection system employed a gyroscope to measure the angular velocity of the foot and three force sensitive resistors to assess the forces exerted by the foot on the shoe sole during walking. A rule-based detection algorithm, which was running on a portable microprocessor board, processed the sensor signals. In the presented experimental study ten able body subjects and six subjects with impaired gait tested the device in both indoor and outdoor environments (0-25 degrees C). The subjects were asked to walk on flat and irregular surfaces, to step over small obstacles, to walk on inclined surfaces, and to ascend and descend stairs. Despite the significant variation in the individual walking styles the system achieved an overall detection reliability above 99% for both subject groups for the tasks involving walking on flat, irregular, and inclined surfaces. In the case of stair climbing and descending tasks the success rate of the system was above 99% for the able body subjects and above 96 % for the subjects with impaired gait. The experiments also showed that the gait phase detection system, unlike other similar devices, was insensitive to perturbations caused by nonwalking activities such as weight shifting between legs during standing, feet sliding, sitting down, and standing up.