Substantia Nigra Pars Reticulata Neurons Research Articles

Involvement of N-methyl- d-aspartate receptor in the delayed transneuronal regression of substantia nigra neurons in rats

The substantia nigra pars reticulata (SNr) receives both inhibitory GABAergic and excitatory glutamatergic afferents from diverse origins. Ischemic injury to the striatum and/or the globus pallidus causes delayed transneuronal death of the SNr neurons, in the course of which neuronal disinhibition induced by loss of GABAergic inputs is supposed to trigger a lethal hypermetabolic process. In the in vivo experiment presented herein, we clarified the role of glutamatergic action via the N-methyl- d-aspartate receptor in this cell death process. Continuous intraventricular infusion (0.5 μl/h) of the N-methyl- d-aspartate receptor antagonist MK-801 (1000 μg/ml), or of saline (control group) was initiated 24 h after 2 h of transient middle cerebral artery (MCA) occlusion in rats, by which massive ischemic injury was produced in the striatopallidal regions. The measured rectal temperature was not significantly altered in the MK-801-infused and in the control rats throughout the time period examined. The rats were killed at 15 days after MCA occlusion. The volume of the focal ischemic infarction of the MK-801-infused group did not significantly differ from that of controls. Also, MK-801-infusion did not significantly ameliorate the nigral atrophy subsequent to MCA occlusion. In association with a marked depletion of GABAergic afferent fibers, neuronal cell number in the ipsilateral SNr was significantly decreased in the control group. In contrast, the neuronal cell loss in the nucleus was completely prevented in the MK-801-infusion group. The data suggested that withdrawal of GABAergic inputs may cause a severe imbalance between excitation and inhibition of the SNr neurons and may eventually result in neurotoxicity mediated by the N-methyl- d-aspartate receptor. Suppression of glutamatergic excitatory effects by suitable drugs may be a reasonable therapy for the transneuronal death of the SNr neurons.

The novel antiepileptic drug levetiracetam (ucb L059) induces alterations in GABA metabolism and turnover in discrete areas of rat brain and reduces neuronal activity in substantia nigra pars reticulata

Levetiracetam (( S)-α-ethyl -2-oxo-pyrrolidine acetamide, ucb L059) is a novel anticonvulsant drug presently in clinical development. Its mechanism of action is unknown although a recently reported novel specific binding site for [ 3H]levetiracetam, unique to brain, may be involved. This binding site has not yet been characterized, but some evidence suggested a possibly indirect interaction with the GABA system. We therefore examined levetiracetam's effects on GABA metabolism and turnover in several rat brain regions after systemic administration of anticonvulsant doses. Furthermore, in order to study functional effects of levetiracetam on a well defined system of GABAergic neurons in a brain region that has been critically involved in anticonvulsant drug action, we examined levetiracetam's action on spontaneous firing of substantia nigra pars reticulata (SNR) neurons in anesthetized rats. Although levetiracetam did not alter the activity of the GABA synthesizing and degrading enzymes glutamic acid decarboxylase (GAD) and GABA aminotransferase (GABA-T) in vitro, systemic administration induced significant alterations in these enzymes in several brain regions, indicating that these enzyme alterations were no direct drug effects but a consequence of postsynaptic changes in either GABAergic or other neurotransmitter-related systems. In the striatum, levetiracetam, 170 mg/kg i.p., induced a significant increase in GABA-T activity while GAD activity markedly decreased. When GABA turnover was estimated after inhibition of GABA-T by aminooxyacetic acid (AOAA), treatment with levetiracetam (given 15 min prior to injection of AOAA) significantly reduced GABA turnover in the striatum. Since the substantia nigra pars reticulata (SNR) receives a strong GABAergic input from the striatum, we examined if the alterations in GABA metabolism and turnover in the striatum led to functional alterations in neuronal activity in the SNR by recording single unit activity of SNR neurons after i.p. injection of levetiracetam. While injection of vehicle did not affect SNR neuronal activity, a significant decrease in spontaneous neuronal firing was recorded after levetiracetam. Since a substantial body of evidence suggests that the SNR is a critical site at which decrease of neuronal firing results in protection against various seizure types, the suppressive effect of levetiracetam on SNR activity may contribute to the anticonvulsant action of this drug.

Turning behavior induced by injections of glutamate receptor antagonists into the substantia nigra of the rat.

We have found recently that muscimol microinjections into the subthalamic nucleus produce contralateral turning activity [Murer and Pazo (1993) NeuroReport, 4:1219-1222]. To test the hypothesis that a reduced glutamate action on substantia nigra pars reticulata neurons mediates this turning response, we examined the effect of unilateral intranigral microinjections of the AMPA/kainate receptor antagonist 6,7-dinitro-quinoxaline-2,3-dione (DNQX) and the competitive N-methyl-D-aspartate (NMDA) receptor antagonist DL-2-amino-5-phosphonovaleric acid (AP-5). DNQX and AP-5 both produced a dose-dependent contralateral turning response, while vehicle administration did not induce turning activity. Application of glutamate receptor antagonists at adjacent regions of the mesencephalic tegmentum were also ineffective. Coadministration of NMDA or AMPA significantly reduced the turning response induced by AP-5 or DNQX, respectively. Lesions of the nigrostriatal pathway by 6-hydroxydopamine did not modify the response to DNQX or AP-5 administration into the nigra. However, their behavioral effects were significantly reduced by a lesion of the ipsilateral subthalamic nucleus. Our results show that the blockade of a tonic input acting on AMPA/kainate and NMDA receptors located at the substantia nigra produces contralateral turning behavior. The effect seems to involve pars reticulata cells since this area remains unchanged after destruction of dopaminergic neurons. The subthalamic nucleus seems to be the endogenous source of the agonist acting on the nigral glutamate receptors related to turning behavior.

Local pressure application of cannabinoid agonists increases spontaneous activity of rat substantia nigra pars reticulata neurons without affecting response to iontophoretically-applied GABA

This study tested the hypothesis that cannabinoid agonists, applied locally into the pars reticulata of substantia nigra (SNpr), could modulate striatonigral transmission, without affecting the response of SNpr neurons to iontophoretically-applied GABA. Multibarreled glass capillary electrode assemblies were used for extracellular recording of the spontaneous electrical activity of single SNpr cells in anesthetized rats. Local pressure ejection of the cannabinoid agonists Win 55212-2 (WIN2) and CP 55940 increased SNpr spontaneous firing rate by 13–46%, similar to the effects of systemic injections. Neither WIN2 nor CP 55940 had an effect on the slowing of SNpr neuron activity in response to iontophoretic GABA. Local pressure application of Win 55212–3 (the much less active enantiomer of WIN2) produced an insignificant decrease in SNpr firing rate. Similarly, locally applied vehicle (45% 2-hydroxypropyl-β-cyclodextrin) produced insignificant decreases in SNpr firing. A second application of cannabinoid agonist produced a much smaller effect, suggesting desensitization. Increasing the interval between CP 55940 applications to 45 min showed recovery of sensitivity to the agonist. Local application of the cannabinoid antagonist, SR 141716A, significantly decreased spontaneous cell firing by 34%. CP 55940, when given immediately following or concurrently with the antagonist application failed to produce the expected increase in discharge rate over baseline. A second application of CP 55940 45 min later produced a 26% increase in firing rate. Bicuculline methiodide (BMI) was applied locally causing a significant increase in SNpr cell firing. CP 55940, when locally administered concurrently with bicuculline methiodide, had no further effect on the firing rate of the cell. Based on the reported presynaptic localization of cannabinoid receptors in SNpr, these findings suggest that cannabinoids act within the SNpr to modulate striatonigral neurotransmission presynaptically. The effect of SR 141716A suggests that an endogenous cannabinoid may mediate striato-nigral transmission.

Amphetamine-induced release of dendritic dopamine in substantia nigra pars reticulata: D1-mediated behavioral and electrophysiological effects.

Dopamine (DA) released from dendrites of substantia nigra dopaminergic neurons potentially is in a position to modulate basal ganglia outputs from the substantia nigra pars reticulata (SNR) via stimulation of D1 receptors on the terminals of striatonigral afferents. The effects of endogenous DA release in the SNR were examined in rats using behavioral activation, multiunit activity of SNR neurons, and cortical EEG pattern as dependent measures. Unilateral infusion of amphetamine (AMPH) into SNR (10 micrograms/0.5 microliter; 5 min) produced a short-lasting behavioral activation that was blocked by coinfusion of the D1 DA receptor antagonist SCH 23390 (0.5 micrograms). Multiunit recordings of SNR neurons in anesthetized rats showed that AMPH, infused as above, produced a rapid decrease in SNR activity. This decrease was maximal (approximately 90%) during the first 10 min postinfusion, followed by a gradual return to baseline levels. Coinfusion of SCH 23390 blocked the AMPH-induced decrease in SNR activity, although by itself this drug produced a 40% decrease in activity. Cortical EEG acquired during the SNR infusions/recordings showed a short-duration change in pattern immediately after AMPH infusion. A relative shift in power from the lowest frequency interval determined (0.8-2.7 Hz) to the next higher frequency interval (2.7-6.8 Hz) was observed which could be prevented by coinfusion of SCH 23390. Thus, dendritically released DA can inhibit the activity of SNR neurons via local stimulation of D1 receptors. This effect is associated with a brief behavioral activation and EEG desynchronization.

Electrophysiological investigation of adenosine trisphosphate-sensitive potassium channels in the rat substantia nigra pars reticulata

Adenosine trisphosphate-sensitive potassium (K-ATP) channels in the substantia nigra pars reticulata were studied in rat brain slices using whole-cell patch clamp recording. Substantia nigra pars reticulata neurons were identified as such by their spontaneous action potential firing at mean rate of 15.3 Hz, virtual absence of hyperpolarization-activated inward current I h, and unresponsiveness to dopamine (30 μM), quinpirole (10 μM) and [Met]enkephalin (10 μM). Intracellular dialysis with Mg 2+-ATP-free pipette solutions caused a slowly developing membrane hyperpolarization (13±4 mV), accompanied by a cessation of action potential firing, or an outward current (79±30 pA at around −60 mV), which were reversed by the sulphonylurea K-ATP channel blockers tolbutamide (100 μM) and glibenclamide (3 μM). When Mg 2+-ATP (2 mM) was included in the recording pipette no membrane hyperpolarization or outward current was observed. Neither the sulphonylureas nor the potassium channel activator lemakalim (200 μM) altered membrane potential, firing rate or holding current under these recording conditions. The outward current induced by dialysis with Mg 2+-ATP-free solutions reversed polarity negative to −94±9 mV (9 cells), close to the estimated K + equilibrium potential (−105 mV) for the conditions used, and was associated with a conductance increase that was blocked by Ba 2+ (100 μM). The current blocked by the sulphonylureas had a similar reversal potential (−97±7 mV; 13 cells), and both currents were voltage independent over the range −50 to −100 mV with slope conductance of approximately 2.0 nS. Outward synaptic currents were evoked by single shock electrical stimulation, in the presence of glutamate receptor antagonists, at a holding potential of −50 mV. These synaptic currents were blocked by bicuculline (10 μM) and reversed polarity at around −65 mV, close to the Cl − equilibrium potential, and were thus mediated by GABA A receptors. They were reversibly depressed by 37±14% in lemakalim (200 μM) in 6/12 cells tested, an effect that was partially reversed by tolbutamide (200 μM). It is concluded that functional K-ATP channels are present both presynaptically and postsynaptically in the substantia nigra pars reticulata. Postsynaptic K-ATP channels may control excitability in conditions where intracellular ATP is reduced, whereas presynaptic K-ATP channels, sensitive to the potassium channel activator lemakalim, can modulate the release of GABA, which probably arises from fibres of extranigral origin. Pharmacological differences between these two sites could be exploited to treat epilepsies, dyskinesias and akinesia.

Responses of substantia nigra pars reticulata neurons to intrastriatal D1 and D2 dopaminergic agonist injections in the rat

Extracellular neuronal recordings were performed in the substantia nigra pars reticulata (SNr) of normal and 6-hydroxydopamine (6-OHDA)-lesioned rats during intrastriatal D1 and D2 dopaminergic-agonist injections. Three types of responses were observed, inhibition, excitation and/or regularization of neuronal discharge patterns. In normal rats, variable responses were observed after D2 injections and a predominance of inhibition after D1 injections. In lesioned rats, the percentage of neurons inhibited and that of neurons regularized increased following D2 injections. During sequential intrastriatal D1 and D2 agonist injections, a response to both dopaminergic agents was observed in 22.5% and 55.5% of SNr neurons in normal and lesioned rats, respectively. Our data suggests that the physiological relevance of direct and indirect pathways convergence upon a given SNr target neuron depends on their respective synaptic weight, the influence of surrounding SNr neurons and the state of dopamine depletion.

(−)-Stepholidine acts as A D 1 partial agonist on firing activity of substantia nigra pars reticulata neurons in 6-hydroxydopamine-lesioned rats

(−)-Stepholidine (SPD), a novel dopamine (DA) D, and/or D 2 receptor antagonist in normosensitive animals, shows agonistic effects on D 1 receptors in rotational behavior of 6-hydroxydopamine (6-OHDA)-lesioned rats. To further characterize the pharmacological properties of SPD, we investigated the effects of SPD on firing activity of substantia nigra pars reticulata (SNR) neurons in different sensitive models. In control rats, the selective D 1 agonist SKF38393 (4 mg/kg, i.v.) induced inconsistent changes (i.e. increase, decrease or no change) in firing of SNR neurons. These effects were completely antagonized by SPD (i.v.), regardless of the changes induced by SKF38393. SPD (4 mg/kg), per se, increased firing by 30.9 ± 14.4%. In reserpinized rats, SKF38393 also induced SPD-reversible inconsistent changes as in control rats. Nevertheless, SPD per se produced no alteration in firing of SNR neurons. In 6-OHDA-lesioned rats, 5 6 SNR neurons were inhibited by SKF38393. The inhibition was completely abolished by Sch23390, a selective D 1 antagonist (0.5–2 mg/kg), but partially reversed by SPD (1–16 mg/). Moreover, SPD (4 mg/kg) itself caused SNR increased or decreased neuron firing, and these effects were completely reversed by Sch23390 (0.5–2 mg/kg) in 8 12 neurons recorded. These results suggest that SPD acts as a partial agonist to D 1 receptors in 6-OHDA-lesioned rats, but as an antagonist to D 1, receptors in normal and reserpinized rats.

Total number of neurons in the neostriatal, pallidal, subthalamic, and substantia nigral nuclei of the rat basal ganglia: a stereological study using the cavalieri and optical disector methods.

The total number of neurons within six subdivisions of the rat basal ganglia was estimated using unbiased stereological counting methods and systematic random sampling techniques. Six young adult rats were perfuse-fixed, their right cerebral hemispheres were embedded in glycolmethacrylate, and a complete set of serial 40-mu m sections was cut through each hemisphere. After a random start, a systematic subset (e.g., every tenth) of these sections was used to estimate the total volume of each subdivision using Cavalieri's method. The same set of sampled sections was used to estimate the number of neurons in a known subvolume (i.e., the Nv) by the optical disector method. The product of the total volume and the Nv by these methods yields an unbiased estimate of the total number of neurons. It was found that the right basal ganglia consisted, on average, of 2.79 million neostriatal or caudate-putamen neurons (with a coefficient of variation of 0.07), 46,000 external globus pallidus neurons (0.11), 3,200 entopeduncular/internal globus pallidus neurons (0.10), 13,600 subthalamic neurons (0.10), 7,200 substantial nigra pars compacta neurons (0.15), and 26,300 substantia nigra pars reticulata neurons (0.07).

Prefrontal cortex inputs of the nucleus accumbens-nigro-thalamic circuit.

The functional organization of the cortico-nucleus accumbens-substantia nigra pars reticulata circuit was investigated in the rat using combined anatomical and electrophysiological approaches. The nucleus accumbens neurons which project to the substantia nigra pars reticulata are located in a circumscribed region of the core immediately adjacent and extending dorsally to the anterior commissure. As shown by retrograde and anterograde transports of wheatgerm agglutinin conjugated to horseradish peroxidase, the region of the nucleus accumbens related to the substantia nigra was found to receive bilateral inputs from restricted areas of the medial and lateral prefrontal cortex, i.e., prelimbic/medial orbital and dorsal agranular insular areas. The electrical stimulation of these medial and lateral prefrontal cortical areas induced excitatory responses in nucleus accumbens neurons projecting to the dorsomedial substantia nigra pars reticulata. Interestingly, an important proportion (61%) of the nucleus accumbens-nigral cells responding to the stimulation of the lateral prefrontal cortex were also excited by the stimulation of the medial prefrontal cortex, demonstrating the existence of a convergent influence of these cortical areas on single nucleus accumbens cells. Furthermore, the present data also show that the stimulation of the medial prefrontal cortex results in a powerful inhibition of the tonic firing of the substantia nigra pars reticulata neurons. In conclusion, this study reveals the existence of a functional link between the prefrontal cortex (prelimbic/medial orbital and agranular insular areas) and the nucleus accumbens neurons which innervate the dorsomedial region of the substantia nigra pars reticulata. Since the dorsomedial region of substantia nigra pars reticulata is known to project to subfields of the mediodorsal and ventromedial thalamic nuclei related to the prefrontal cortex, the present data further demonstrate the existence of a prefrontal-nucleus accumbens-thalamo-cortical circuit involving the substantia nigra pars reticulata.

Electrophysiological changes in substantia nigra after striatal infarction

The GABAergic efferent pathway from the striatum exerts inhibitory control on the substantia nigra pars reticulata (SNR) neurones. We studied sequential changes in spontaneous single-unit activities in the ipsilateral SNR 1 h, 1 day, 7 days and 14 days after striatal infarction induced by middle cerebral artery occlusion (MCAO) in rats. Compared with a sham-operated group, there was no change in the firing rate 1 h after MCAO, and one day after MCAO the mean firing rate decreased. The firing rate of SNR neurones at 7 and 14 days after MCAO was significantly reduced compared with the sham-operated group. Only two neurones (sham-operated group and 1 h after MCAO group) fired with high frequency. Histological examination revealed degeneration of the ipsilateral SNR 7 and 14 days after MCAO. Our results indicated that SNR neuronal degeneration accompanied by striatal ischaemia does not simply depend on hyperexcitation due to the activation of a disinhibition mechanism.

Electrophysiological changes in substantia nigra after striatal infarction

The GABAergic efferent pathway from the striatum exerts inhibitory control on the substantia nigra pars reticulata (SNR) neurones. We studied sequential changes in spontaneous single-unit activities in the ipsilateral SNR 1 h, 1 day, 7 days and 14 days after striatal infarction induced by middle cerebral artery occlusion (MCAO) in rats. Compared with a sham-operated group, there was no change in the firing rate 1 h after MCAO, and one day after MCAO the mean firing rate decreased. The firing rate of SNR neurones at 7 and 14 days after MCAO was significantly reduced compared with the sham-operated group. Only two neurones (sham-operated group and 1 h after MCAO group) fired with high frequency. Histological examination revealed degeneration of the ipsilateral SNR 7 and 14 days after MCAO. Our results indicated that SNR neuronal degeneration accompanied by striatal ischaemia does not simply depend on hyperexcitation due to the activation of a disinhibition mechanism.

Excitation of rat substantia nigra pars reticulata neurons by 5-hydroxytryptaminein vitro: Evidence for a direct action mediated by 5-hydroxytryptamine2C receptors

Single-unit extracellular and whole-cell patch clamp recording were used to study the actions of exogenously applied 5-hydroxytryptamine on substantia nigra pars reticulata neurons in parasaggital slices of rat midbrain. Seventy-six per cent of substantia nigra pars reticulata cells (254/334) recorded extracellularly were excited by 5-hydroxytryptamine (EC50 = 9.56 microM); in the remainder, inhibitions (13.5%), biphasic responses (4.2%) or lack of response (6.3%) were observed. Using whole-cell patch recording, 5-hydroxytryptamine (10 microM) caused either an inward current (9/9 cells) or a depolarization (3/3 cells) at membrane potentials in the range -50 to -90 mV, which was resistant to tetrodotoxin (4/4 cells), indicating that the predominant, excitatory action of 5-hydroxytryptamine was due to a direct action on substantia nigra pars reticulata neurons. The 5-hydroxytryptamine excitation (recorded extracellularly) was reduced to 24 +/- 6% of control values by methysergide (0.1 microM) and to 17 +/- 5% of control by ketanserin (10 microM), but was unaffected by the 5-hydroxytryptamine antagonists spiperone (0.1 microM), yohimbine (0.1 microM), pindolol (1 microM), GR113808A (1 microM) or ICS 205930 (10 microM). In addition, the 5-hydroxytryptamine excitation was mimicked by the 5-hydroxytryptamine2C receptor--preferring agonist alpha-methyl 5-hydroxytryptamine (10 microM), but the agonists CP93, 129 (0.1-1 microM) and (+/-)-2-dipropylamino-8-hydroxy-1,2,3,4-tetrahydronaphthalene hydrobromide (0.1-1 microM) were without effect. Taken together, this pharmacology indicated involvement of the 5-hydroxytryptamine2C receptor in the 5-hydroxytryptamine excitation, while other candidate receptors known to be present in rat substantia nigra pars reticulata (5-hydroxytryptamine1B, 5-hydroxytryptamine2A and 5-hydroxytryptamine4) could be excluded from consideration. While in accord with current information on the location of 5-hydroxytryptamine receptor subtypes in substantia nigra pars reticulata, and the consequence of activation of neuronal 5-hydroxytryptamine2C receptors, these results contrast with data from in vivo experiments which suggest that the net effect of 5-hydroxytryptamine is to inhibit substantia nigra pars reticulata neurons. The reason for this apparent discrepancy may lie in detailed consideration of the microcircuitry of the substantia nigra pars reticulata. This may lead to a re-evaluation of the influence of 5-hydroxytryptamine on this basal ganglia output relay nucleus, and its role in motor control and the gating of generalized seizure activity.

Reduction in firing rate of substantia nigra pars reticulata neurons by valproate: influence of different types of anesthesia in rats

Nondopaminergic, presumably GABAergic neurons in the substantia nigra pars reticulata (SNR) are thought to function as a gating mechanism for seizure propagation. Systemic administration of anticonvulsant doses of the antiepileptic drug valproate (VPA) has previously been reported to inhibit the firing of nondopaminergic SNR neurons in anesthetized but not in awake, paralyzed and locally anesthetized rats, suggesting that the findings in anesthetized rats were due to an interaction between VPA and the general anesthetic used. In the present study, we determined the influence of different anesthetic measures on the effect of an anticonvulsant dose of VPA (100 mg/kg) on extracellularly recorded spontaneous single unit activity of nondopaminergic SNR neurons in rats. Rats were anesthetized by continuous infusion of the general anesthetic chloral hydrate, the dissociative anesthetic ketamine or the narcotic opioid fentanyl, or were only locally anesthetized and paralyzed. VPA significantly reduced SNR firing in all groups with a time course that matched its anticonvulsant time course in rodents. However, VPA's inhibitory effect on SNR firing was significantly less marked under anesthesia with chloral hydrate than in any of the other groups, indicating that this anesthetic suppresses the action of VPA, which may be related to an interaction with GABA-related processes in the SNR. The closest approximation to the effect of VPA in awake rats was obtained under anesthesia with ketamine, while VPA's inhibitory action on SNR neuronal firing seemed to be enhanced in the fentanyl group, which exhibited the highest baseline firing rates of all groups. Determination of VPA in the SN showed that the difference in VPA's inhibitory effect on SNR neurons was not secondary to differences in local drug concentrations. The data demonstrate that VPA is capable of significantly slowing the spontaneous activity of nondopaminergic SNR neurons, but that the magnitude of this effect depends on the anesthetic measures used. In view of the presumed role of SNR neurons in seizure propagation and the finding that VPA consistently inhibits these neurons at an anticonvulsant dose, the present data suggest that suppression of spontaneous SNR neuronal firing may be an important mechanism through which VPA exerts its anticonvulsant properties.

Reduction of apomorphine-induced rotational behaviour by subthalamic lesion in 6-OHDA lesioned rats is associated with a normalization of firing rate and discharge pattern of pars reticulata neurons.

The effect of subthalamic nucleus (STh) lesion on apomorphine-induced rotational behaviour and unit activity of substantia nigra pars reticulata (SNr) neurons was studied in normal, sham-control and unilateral 6-OHDA-lesioned rats [SN pars compacta (SNc)-lesioned]. In the latter, contraversive rotational behaviour was greatly reduced by an additional ipsilateral STh lesion. A moderate ipsiversive rotation was observed in rats with a single STh lesion. Concurrently, SN unit extracellular recordings were performed in age-matched normal rats, sham-controls for both lesions, STh-lesioned rats, SNc-lesioned rats, and SNc-lesioned rats with an ipsilateral STh lesion (SNc+STh-lesioned). Pars reticulata neurons had a higher mean firing rate in SNc-lesioned rats than in control rats. Furthermore, 68% of SNr neurons in SNc-lesioned rats had a tonic discharge pattern (against 92.3% in control rats) and 32% a mixed or bursting pattern. After STh lesion, a clear decrease in SNr firing rate was observed in SNc-lesioned rats. Moreover, STh lesion improved interspike interval regularity and decreased the occurrence of bursting patterns. In rats with a single STh lesion, the firing rate was no different from that of the sham-controls but the discharge pattern was more regular. These data show that STh lesion decreased apomorphine-induced rotational behaviour in dopamine-depleted animals. This effect could be related to the suppression of the excitatory effect of STh efferents on the SNr neurons. STh lesion both counterbalanced the increased activity of SNr neurons and regularized their discharge pattern.

Effects of a cannabinoid on spontaneous and evoked neuronal activity in the substantia nigra pars reticulata

Single unit electrophysiology was used to explore the role of cannabinoid receptors in the substantia nigra pars reticulata. Intravenous and intraperitoneal injections of the potent and selective synthetic cannabinoid ( R)-(+)-[2,3-dihydro-5-methyl-3-[(4-morpholinyl)methyl]pyrrolo[1,2,3-de]-1,4-benzoxazin-6-yl](1-naphthalenyl) methanone (WIN 55,212-2) produced modest but significant increases in the spontaneous firing rate of neurons in the substantia nigra pars reticulata. In a second set of experiments, WIN 55,212-2 (up to 1.0 mg/kg i.v.) antagonized the inhibition of firing produced in the substantia nigra pars reticulata by electrical stimulation of the striatum. The pharmacological specificity of this effect was demonstrated using the inactive enantiomer WIN 55,212-3. The possibility that WIN 55,212-2 exerts its effects by regulating γ-aminobutyric acid (GABA) release from striatonigral fibers was suggested by the observation that bicuculline (up to 0.5 mg/kg i.v.) reversed the effect of striatal stimulation. It thus appears that cannabinoid receptors on striatonigral neuron terminals may regulate movement by disinhibiting the activity of substantia nigra pars reticulata neurons, perhaps by inhibiting the release of GABA into the substantia nigra pars reticulata.

Gramicidin‐perforated patch recording: GABA response in mammalian neurones with intact intracellular chloride.

1. By the development of a new perforated patch method using gramicidin, the effects of GABA on neurones dissociated from the rat substantia nigra pars reticulata (SNR) were examined without disturbing the intracellular chloride concentration. 2. Using the patch pipette solution containing gramicidin (100 micrograms ml-1), the access resistance dropped to less than 20 M omega within 40 min after making the gigaohm seal. 3. Under current-clamp conditions, GABA caused a hyperpolarization accompanied by a blockade of spontaneous firing. Under voltage clamp at a holding potential (Vh) of -50 mV, GABA evoked an outward current by way of bicuculline- and picrotoxin-sensitive GABAA receptors. 4. A 10-fold change of extracellular chloride concentration resulted in a 58 mV shift of the reversal potential of GABA-induced outward current (EGABA), indicating that the membrane behaves like a chloride electrode in the presence of GABA. 5. The intracellular chloride activities (aCli), calculated with the Nernst equation using both extracellular chloride activity and EGABA values, ranged from 2.8 to 19.7 mM with a mean value of 9.5 mM. The aCli was not affected either by different pipette solutions or by different holding potentials more hyperpolarized than -40 mV. 6. In the recording from SNR neurones in brain slice using the gramicidin-perforated patch-clamp technique, the inhibitory and excitatory postsynaptic currents were recorded in different current directions and the former was blocked by bicuculline. 7. In conclusion, the gramicidin-perforated patch method will disclose previously unknown aspects of biological responses involving Cl-.

Demonstration of axon collateral projections from the substantia nigra pars reticulata to the superior colliculus and the parvicellular reticular formation in the rat

It was revealed in the rat that single neurons in the substantia nigra pars reticulata (SNr) innervated both the superior colliculus (SC) and the parvicellular reticular formation (RFp) in the pons and medulla oblongata by way of axon collaterals. After injecting Fluoro-gold into the lateral part of the SC and Fluoro-ruby into the RFp on the same side, some SNr neurons were double-labeled with both tracers. They were localized in the dorsolateral part of the caudal half of the SNr ipsilateral to the injection sites.

Occurrence of heat shock response in deafferented neurons in the substantia nigra of rats

The substantia nigra is innervated by massive inhibitory GABAergic projections from the striatum and globus pallidus, deafferentation of which is supposed to lead to anterograde trans-synaptic degeneration of the nigral neurons. An immunohistochemical method was used to examine the induction of 72,000 mol. wt heat shock protein in the substantia nigra following cerebral hemitransection or transient middle cerebral artery occlusion. At three and four days post-transection, strong immunoreactivity for 72,000 mol. wt heat shock protein was found in the ipsilateral substantia nigra pars reticulata. Light microscopic observation revealed a number of pars reticulata neurons showing strong immunoreactivity for 72,000 mol. wt heat shock protein in their perikarya and proximal processes. In addition, Golgi-like stained neurons with dystrophic features were occasionally observed in the ipsilateral substantia nigra pars reticulata. The immunoreactivity for 72,000 mol. wt heat shock protein in the ipsilateral pars reticulata gradually declined and almost disappeared by 15 days after transection. No apparent induction of 72,000 mol. wt heat shock protein was found in the substantia nigra pars compacta throughout the time period examined. Massive striatal ischémie injury produced by transient middle cerebral artery occlusion also induced expression of 72,000 mol. wt heat shock protein in the pars reticulata neurons three and four days postoperatively. These findings suggest that deafferentation of the striatal or striatopallidal inputs per se is a harmful stress for the substantia nigra pars reticulata neurons, inducing 72,000 mol. wt heat shock protein synthesis. The present data may contribute to our understanding of the molecular basis of the pathomechanism of the transneuronal regression of substantia nigra pars reticulata neurons, which may occur after removal of inhibitory GABAergic inputs.

Rat substantia nigra pars reticulata neurones are tonically inhibited via GABA A, but not GABA B, receptors in vitro

Extracellular single unit recordings were made from substantia nigra pars reticulata (SNr) neurones in slices of rat brain. Cells fired spontaneous action potentials at 11.4 ± 0.8Hz. The GABA A receptor agonist isoguvacine (1–10 μM) reduced firing rate in a concentration-dependent manner [50% of maximal inhibition (IC 50) with 3.2 μM], as did the GABA B agonist baclofen (0.3–10 μM; IC 50 1.4 μM). The GABA A antagonist bicuculline (30 μM) not only blocked the action of isoguvacine, but also increased the basal firing rate to 187.5 ± 12.6% of control. The GABA B antagonist CGP 55845A (0.1 μM), while blocking the inhibitory action of baclofen, was without effect on spontaneous firing rate, as was strychnine (10 μM), the antagonist of glycine and taurine, and also Met-enkephalin (10 μM). Tiagabine (50 μM), the blocker of GABA uptake, caused an inhibition of firing which could be reversed with bicuculline (30 μM) but not CGP 55845A (1 μM). We conclude that the firing rate of SNr neurones is under tonic inhibition by GABA in vitro, which can be relieved by antagonists of GABA A, but not GABA B receptors, and enhanced by blockade of GABA reuptake. The source of this GABA tone is likely to be from recurrent axon collaterals of SNr neurones themselves.