Reticulata Neurons Research Articles

Dopaminergic Presynaptic Modulation of Nigral Afferents: Its Role in the Generation of Recurrent Bursting in Substantia Nigra Pars Reticulata Neurons

Previous work has shown the functions associated with activation of dopamine presynaptic receptors in some substantia nigra pars reticulata (SNr) afferents: (i) striatonigral terminals (direct pathway) posses presynaptic dopamine D1-class receptors whose action is to enhance inhibitory postsynaptic currents (IPSCs) and GABA transmission. (ii) Subthalamonigral terminals posses D1- and D2-class receptors where D1-class receptor activation enhances and D2-class receptor activation decreases excitatory postsynaptic currents. Here we report that pallidonigral afferents posses D2-class receptors (D3 and D4 types) that decrease inhibitory synaptic transmission via presynaptic modulation. No action of D1-class agonists was found on pallidonigral synapses. In contrast, administration of D1-receptor antagonists greatly decreased striatonigral IPSCs in the same preparation, suggesting that tonic dopamine levels help in maintaining the function of the striatonigral (direct) pathway. When both D3 and D4 type receptors were blocked, pallidonigral IPSCs increased in amplitude while striatonigral connections had no significant change, suggesting that tonic dopamine levels are repressing a powerful inhibition conveyed by pallidonigral synapses (a branch of the indirect pathway). We then blocked both D1- and D2-class receptors to acutely decrease direct pathway (striatonigral) and enhance indirect pathways (subthalamonigral and pallidonigral) synaptic force. The result was that most SNr projection neurons entered a recurrent bursting firing mode similar to that observed during Parkinsonism in both patients and animal models. These results raise the question as to whether the lack of dopamine in basal ganglia output nuclei is enough to generate some pathological signs of Parkinsonism.

Functional heterogeneity of NMDA receptors in rat substantia nigra pars compacta and reticulata neurones

The nigra substantia nigra pars compacta (SNc) and substantia pars reticulata (SNr) form two major basal ganglia components with different functional roles. SNc dopaminergic (DA) neurones are vulnerable to cell death in Parkinson's disease, and NMDA receptor activation is a potential contributing mechanism. We have investigated the sensitivity of whole-cell and synaptic NMDA responses to intracellular ATP and GTP application in the SNc and SNr from rats on postnatal day (P) 7 and P28. Both NMDA current density (pA/pF) and desensitization to prolonged or repeated NMDA application were greater in the SNr than in the SNc. When ATP levels were not supplemented, responses to prolonged NMDA administration desensitized in P7 SNc DA neurones but not at P28. At P28, SNr neurones desensitized more than SNc neurones, with or without added ATP. Responses to brief NMDA applications and synaptic NMDA currents were not sensitive to inclusion of ATP in the pipette solution. To investigate these differences between the SNc and SNr, NR2 subunit-selective antagonists were tested. NMDA currents were inhibited by ifenprodil (10 microM) and UBP141 (4 microM), but not by Zn(2+) (100 nm), in both the SNr and SNc, suggesting that SNc and SNr neurones express similar receptor subunits; NR2B and NR2D, but not NR2A. The different NMDA response properties in the SNc and SNr may be caused by differences in receptor modulation and/or trafficking. The vulnerability of SNc DA neurones to cell death is not correlated with NMDA current density or receptor subtypes, but could in part be related to inadequate NMDA receptor desensitization.

MPTP treatment increases expression of pre-pro-nociceptin/orphanin FQ mRNA in a subset of substantia nigra reticulata neurons

Antagonists selectively inhibiting activation of the nociceptin/orphanin FQ (N/OFQ) receptor reduce motor symptoms in experimental models of Parkinson's disease, and genetic deletion of the ppN/OFQ gene offers partial protection of mid-brain dopamine neurons against the neurotoxin, 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). MPTP increased ppN/OFQ mRNA expression in the substantia nigra (SN). We have evaluated the temporal relationship of dopamine cell loss to increased ppN/OFQ mRNA expression in the substantia nigra after MPTP treatment, and characterized the cellular locations in which increased ppN/OFQ mRNA expression was observed after MPTP treatment. MPTP increased by about 5-fold the number of neurons expressing ppN/OFQ mRNA in the pars reticulata of SN (SNr) by 24 h after treatment and the elevation remained significant for at least 7 days. This period coincided with the timing of the loss of dopamine neurons from the pars compacta of substantia nigra (SNc) after MPTP. The increased expression of ppN/OFQ mRNA co-localized with a neuronal marker in the SNr. MPTP treatment resulted in a small increase in the numbers of neurons expressing ppN/OFQ in the SNc in mice from one mouse colony but the increase did not reach statistical significance in mice from another colony. No changes in ppN/OFQ-mRNA expression were observed in the ventral tegmental area (VTA), the caudate-putamen, the subthalamic nucleus, or in two other brains areas. These results demonstrate that increased N/OFQ expression in the SNr is closely associated with the MPTP-induced loss of dopamine neurons in the SNc in a widely used animal model of Parkinson's disease.

Subthalamic nucleus evokes similar long lasting glutamatergic excitations in pallidal, entopeduncular and nigral neurons in the basal ganglia slice

The subthalamic nucleus (STN) modulates the activity of globus pallidus (GP), entopeduncular nucleus (EP) and substantia nigra pars reticulata (SNr) neurons via its direct glutamatergic projections. To investigate the mechanism by which STN affects activity in these structures and whether STN induced activity is comparable among STN target neurons, we performed patch clamp recordings in a tilted, parasagittal, basal ganglia slice (BGS) that preserves these functional connections. We report that single, brief stimulation of the STN generates a brief monosynaptic AMPA-mediated excitatory postsynaptic current (EPSC) in GP, EP and SNr neurons. A higher intensity, supra-threshold activation evokes a compound EPSC consisting of an early monosynaptic component followed by a slow inward NMDA-mediated current with an overlying barrage of AMPA-mediated EPSCs. These late EPSCs were polysynaptic and gave rise to bursts of spikes that lasted several hundreds of milliseconds. They were eliminated by surgical removal of the STN from the BGS slice, indicating that the STN is required for their generation. Reconstruction of biocytin-filled STN neurons revealed that a third of STN neurons project intra-STN axon collaterals that may underlie polysynaptic activity. We propose that activation of the STN yields comparable long lasting excitations in its target neurons by means of a polysynaptic network.

Noradrenergic lesion of the locus coeruleus increases apomorphine-induced circling behavior and the firing activity of substantia nigra pars reticulata neurons in a rat model of Parkinson's disease

The role of noradrenergic depletion of the locus coeruleus (LC) in the pathophysiology of Parkinson's disease (PD) is still unclear. In the present study, apomorphine-induced circling behavior and extracellular firing activity of substantia nigra pars reticulata (SNr) neurons were examined in rats with unilateral 6-hydroxydopamine lesions of the LC, substantia nigra pars compacta (SNc) and with combined SNc and LC lesions. A moderate contralateral circling was observed in rats with LC lesions after apomorphine. Moreover, the circling behavior was obviously increased by further lesions of LC in SNc-lesioned rats. Extracellular recordings indicated that the firing rate of SNr neurons increased significantly and the firing pattern of these neurons also changed towards more irregular and bursty after SNc lesioning as compared to sham-lesioned rats, while the firing rate and pattern were unaffected in rats with simple lesions of the LC. However, the firing rate of SNr neurons in rats with combined LC and SNc lesions increased significantly when compared to that of rats with simple lesions of the SNc, although the firing pattern was not altered. Furthermore, SNc lesions in rats increased the firing rate of SNr neurons with irregular firing pattern, and additional LC lesions in SNc-lesioned rats increased the firing rate of SNr neurons with regular and irregular firing pattern. These results indicate that lesions of the LC intensify apomorphine-induced circling behavior and lead to a further hyperactivity of SNr neurons in a rat model of PD, suggesting that LC-noradrenergic system is involved in the motor dysfunction of PD.

Cocaine‐induced stereotypy is linked to an imbalance between the medial prefrontal and sensorimotor circuits of the basal ganglia

The dysfunction of basal ganglia circuits related to stereotyped motor activity was analysed using the well-established model of cocaine-induced stereotypy in the rat. We examined and compared the neurochemical and electrophysiological effects occurring in medial prefrontal and sensorimotor basal ganglia circuits of the dorsal striatum after cocaine injection in sensitized and non-sensitized rats. Acute injections of cocaine (25 mg/kg), not inducing stereotyped behaviour, affected both medial prefrontal and sensorimotor circuits in a similar way: (i) a mild and delayed increase and decrease of N-methyl-D-aspartate-evoked dopamine and acetylcholine release, respectively and (ii) a marked decrease of cortically evoked inhibition of substantia nigra pars reticulata neurons revealing an imbalance of information transmission between the direct and indirect trans-striatal pathways. In contrast, following sensitization to cocaine, a challenge injection of the same dose of cocaine, generating strong stereotyped behaviour, provoked neurochemical and electrophysiological effects only in the medial prefrontal but not in the sensorimotor circuits: (i) a strong increase of dopamine and decrease of acetylcholine release in the medial prefrontal territory of the dorsal striatum and (ii) a reduction of all inhibitory and excitatory components of the responses evoked in substantia nigra pars reticulata by medial prefrontal stimulation. Therefore, these data disclose distinct reactivity of the medial prefrontal and sensorimotor circuits of the basal ganglia to repeated cocaine administration leading to stereotyped behaviour induced by subsequent cocaine challenge. Thus, we suggest that stereotyped behaviour is correlated to an imbalance between the medial prefrontal and sensorimotor circuits of the basal ganglia resulting in a loss of control of motor behaviour.

Nitric oxide- and cGMP-active compounds affect the discharge of substantia nigra pars reticulata neurons: in vivo evidences in the rat

The nitric oxide (NO)-active drugs influence on the bioelectric activity of neurons of the pars reticulata of the substantia nigra was studied in urethane-anesthetized rats. A first group of animals was treated with 7-nitro-indazole (7-NI), a preferential inhibitor of neuronal NO synthase. In a second group of rats, electrophysiological recordings were coupled with microiontophoretic administration of Nomega-nitro-L-arginine methyl ester (L-NAME, a NO synthase inhibitor), 3-morpholino-sydnonimin-hydrocloride (SIN-1, a NO donor) and 8-Br-cGMP (a cell-permeable analogue of cGMP, the main second-messenger of NO neurotransmission). 7-NI and L-NAME caused a statistically significant decrease in the firing rate of most of the responsive cells, while application of SIN-1 and 8-Br-CGMP induced statistically significant excitatory effects. The results suggest a NO mediated excitatory modulation of the SNr neurons activity with a possible involvement of the cGMP pathway.

Regular firing of a single output neuron reduces its own inhibition through endocannabinoids in substantia nigra pars reticulata of juvenile mice

Depolarization-induced suppression of inhibition in substantia nigra pars reticulata suggests that burst-like activity but not regular firing suffices to activate presynaptic endocannabinoid CB1 receptors. To more closely determine the type of activity required, we applied gramicidin perforated patch recording under visual control to substantia nigra slices of juvenile mice. We found that evoked inhibitory postsynaptic currents (eIPSCs) were reduced in amplitude by the spontaneous firing of a neuron under study, whereas silencing this neuron enhanced inhibitory responses. Autonomous firing reduced eIPSCs to 78%±2% in a time- but not frequency-dependent manner. The phenomenon which we termed firing-induced suppression of inhibition was cannabinoid receptor subtype 1–dependent, whereas adenosine A1 receptors played only a minor role. Depletion of intracellular Ca2+ stores abolished the firing-induced suppression of inhibition suggesting that Ca2+ release from internal stores is necessary for the production of endocannabinoids during autonomous firing. We suggest that the Ca2+ influx during autonomous activity of pars reticulata neurons suffices to selectively dampen incoming inhibition from striatal neurons because it is amplified by ryanodine receptor-mediated Ca2+ release from intracellular stores.

Synaptic plasticity, dopamine and Parkinson's disease: one step ahead

The progressive loss of substantia nigra pars compacta neurons that characterizes Parkinson's disease pathology leads to impaired levels of dopamine in several key structures of the basal ganglia neuronal circuit and subsequent alteration of the thinly regulated balance between activities of the output nuclei—the internal segment of the globus pallidus and the substantia nigra pars reticulata —that is considered essential for normal function of the circuit (Fig. 1A). Fig. 1 The basal ganglia circuit. Potential implications of the LTP of GABAergic signals onto substantia nigra pars reticulata neurons. ( a) During physiological conditions the thinly regulated activity of the direct and indirect pathways controls the activity of the output nuclei. ( b ) During Parkinson's disease, in the absence of a pharmacological treatment, dopamine deficiency causes overactivity of the indirect pathway and reduced activity of the inhibitory GABAergic direct pathway, disinhibiting the output nuclei and thus causing excessive inhibition of the motor thalamus. In this condition, high frequency stimulation does not induce a lasting change in GABAergic field potential amplitude in the substantia nigra pars reticulata . ( c ) After the administration of l-DOPA, the same high-frequency stimulation protocol induces potentiation of the GABAergic field potential amplitudes, reducing the excessive inhibitory activity that the output nuclei exert on the motor thalamus. ( d ) Finally, after the chronic administration of l-DOPA, uncontrolled potentiation of the GABA-mediated inhibition of the output nuclei might result in long-term suppression of their firing rate which in turn might lead to pathological disinhibition of the thalamus and the subsequent onset of pathological hyperkinetic behaviours, such as l-DOPA-induced dyskinesias. Note that, in the Figure, inhibitory GABAergic connections are represented in red, excitatory glutamatergic connections in green and dopaminergic connections in black. Although many advances have been made in the field of Parkinson's disease research, the precise mechanisms leading to symptom …

Complex EPSCs evoked in substantia nigra reticulata neurons are disrupted by repetitive stimulation of the subthalamic nucleus

Although substantia nigra reticulata (SNR) neurons fire bursts of action potentials during normal movement, excessive burst firing correlates with symptoms of Parkinson's disease. A major excitatory output from the subthalamic nucleus (STN) to the SNR is thought to provide the synaptic impetus for burst firing in SNR neurons. Using patch pipettes to record from SNR neurons in rat brain slices, we found that a single electrical stimulus delivered to the STN evokes a burst of action potentials. Under voltage-clamp conditions, STN stimulation evokes a complex EPSC that is comprised of an initial monosynaptic EPSC followed by a series of late EPSCs superimposed on a long-lasting inward current. Using varied stimulation frequencies, we found that the initial EPSC was significantly reduced or abolished after 2 s of 50-100 Hz STN stimulation. However, only 4 s of 1 Hz stimulation was required to abolish the late component of the complex EPSC. We suggest that differential effects of repetitive STN stimulation on early and late components of complex EPSCs may help explain the frequency-dependent effects of deep brain stimulation of the STN that is used in the treatment of Parkinson's disease.

Differential localization of GABAA receptor subunits within the substantia nigra of the human brain: An immunohistochemical study

Gamma-aminobutyric acid(A) (GABA(A)) receptors (GABA(A)R) are inhibitory heteropentameric chloride ion channels comprising a variety of subunits and are localized at postsynaptic sites within the central nervous system. In this study we present the first detailed immunohistochemical investigation on the regional, cellular, and subcellular localisation of alpha(1), alpha(2), alpha(3), beta(2,3), and gamma(2) subunits of the GABA(A)R in the human substantia nigra (SN). The SN comprises two major regions, the SN pars compacta (SNc) consisting of dopaminergic projection neurons, and the SN pars reticulata (SNr) consisting of GABAergic parvalbumin-positive projection neurons. The results of our single- and double-labeling studies demonstrate that in the SNr GABA(A) receptors contain alpha(1), alpha(3), beta(2,3), and gamma(2) subunits and are localized in a weblike network over the cell soma, dendrites, and spines of SNr parvalbumin-positive nonpigmented neurons. By contrast, GABA(A)Rs on the SNc dopaminergic pigmented neurons contain predominantly alpha(3) and gamma(2) subunits; however there is GABA(A)R heterogeneity in the SNc, with a small subpopulation (6.5%) of pigmented SNc neurons additionally containing alpha(1) and beta(2,3) GABA(A)R subunits. Also, in the SNr, parvalbumin-positive terminals are adjacent to GABA(A)R on the soma and proximal dendrites of SNr neurons, whereas linear arrangements of substance P-positive terminals are adjacent to GABA(A) receptors on all regions of the dendritic tree. These results show marked GABA(A)R subunit hetereogeneity in the SN, suggesting that GABA exerts quite different effects on pars compacta and pars reticulata neurons in the human SN via GABA(A) receptors of different subunit configurations.

Glycine receptors in the striatum, globus pallidus, and substantia nigra of the human brain: An immunohistochemical study

Glycine receptors (GlyRs) are heteropentameric chloride ion channels that facilitate fast-response, inhibitory neurotransmission in the mammalian spinal cord and brain. GlyRs have four functional subunits, alpha1-3 and beta, which likely exist in heteromeric alphabeta combinations. Mutations in GlyR alpha1 and beta subunits are well known for their involvement in hyperekplexia, a paroxysmal motor disorder. In this study we present the first detailed immunohistochemical investigation at the regional, cellular, and subcellular levels of GlyRs in the human basal ganglia. The results show that GlyRs are present at the regional level in low concentrations in the striatum and globus pallidus and are present in the highest concentrations in the substantia nigra. At the cellular level, GlyRs are present only in discrete populations of neurons immunoreactive for choline acetyltransferase (ChAT), parvalbumin, and calretinin in the human striatum, on a subpopulation of parvalbumin- and calretinin-positive neurons in the globus pallidus, and in the substantia nigra GlyRs are present on approximately three-fourths of all pars compacta and one-third of all pars reticulata neurons. They also form a distinct band of immunoreactive neurons in the intermedullary layers of the globus pallidus. At the subcellular level in the substantia nigra pars reticulata (SNr), GlyRs show a localized distribution on the soma and dendrites that partially complements but does not overlap with the distribution of gamma-aminobutyric acid (GABA)A receptors. Our results demonstrate the precise cellular and subcellular localization of GlyRs in the human basal ganglia and suggest that glycinergic receptors may play an important complementary role to other inhibitory receptors in modulating cholinergic, dopaminergic, and GABAergic neuronal pathways in the basal ganglia.

Ca2+ release‐dependent hyperpolarizations modulate the firing pattern of juvenile GABA neurons in mouse substantia nigra pars reticulata in vitro

A phasic activation of small-conductance Ca(2+)-dependent K(+) channels (SK channels) underlies spike-afterhyperpolarizations and spike-independent, transient hyperpolarizations in juvenile substantia nigra neurons. Outward current pulses that cause the spike-independent hyperpolarizations result from ryanodine receptor-mediated Ca(2+) release from intracellular stores. To study the modulation of excitability by the outward current pulses, we recorded from GABAergic pars reticulata neurons of mice at postnatal days 12-16. We induced a prolongation of SK channel open states by 1-ethyl-2-benzimidazolinone (1-EBIO). In addition to a prolongation of spike-afterhyperpolarizations, 1-EBIO (200 microm) potentiated outward current pulses by increasing their duration. Neurons were manipulated by current injection to display continuous or discontinuous discharge. Despite the prolongation of the outward current pulses by 1-EBIO, continuous action potential discharge became more regular, although its frequency declined. Durations of silent periods (periods of >2x average interspike interval) increased. Caffeine (1 mm) further increased the duration of such silent periods. Caffeine, however, had no effect at short interspike intervals (<600 ms). Cyclopiazonic acid (10 microm) silenced discharge in 1-EBIO, but discharge reappeared with the depletion of Ca(2+) stores. We conclude that the modulation of excitability by an activation of SK channels through ryanodine receptor-mediated release of Ca(2+) critically depends on the frequency of discharge. Outward current pulses occur only if interspike intervals exceed the duration of spike-afterhyperpolarizations. In this instance, the phasic, spike-independent activation of SK channels supports pauses to interrupt autonomous discharge in juvenile GABAergic pars reticulata neurons.

Stability of substantia nigra pars reticulata neuronal discharge rates during dopamine receptor blockade and its possible mechanisms

It is hypothesized that substantia nigra pars reticulata neurons become overactive during a deficit of dopamine transmission. In this study, we examined how acute dopamine receptor blockade (SCH23390 and eticlopride) affects impulse activity of substantia nigra pars reticulata neurons and their response to iontophoretic gamma-amino-n-butyric acid in awake, unrestrained rats. No changes in discharge rate were found during complete dopamine receptor blockade, but these neurons showed a diminished response to gamma-amino-n-butyric acid, suggesting gamma-amino-n-butyric acid receptor hyposensitivity. This may result from tonic increase in gamma-amino-n-butyric acid input from the striatum and globus pallidus, which are activated during dopamine receptor blockade. As substantia nigra pars reticulata neurons are autoactive and resistant to tonic increases in gamma-amino-n-butyric acid input, changes in their responsiveness to phasic gamma-amino-n-butyric acid inputs, not tonic increase discharge rate, may underlie movement disturbance following dopamine deficit.

General anesthesia as a factor affecting impulse activity and neuronal responses to putative neurotransmitters

Although it is evident that general anesthesia should affect impulse activity and neurochemical responses of central neurons, there are limited studies in which these parameters were compared in both awake and anesthetized animal preparations. We used single-unit recording coupled with iontophoresis to examine impulse activity and responses of substantia nigra pars reticulata (SNr) neurons to GABA, glutamate (GLU), and dopamine (DA) in rats in awake, unrestrained conditions and during chloral hydrate anesthesia. SNr neurons in both conditions had similar organization of impulse flow, but during anesthesia, they have lower mean rates and discharge variability than in awake conditions. In individual units, discharge rate in awake, quietly resting rats was almost three-fold more variable than during anesthesia. These cells in both conditions were highly sensitive to iontophoretic GABA, but the response was stronger during anesthesia. In contrast to virtually no responses to GLU in awake conditions, most SNr neurons during anesthesia were excited by GLU; the response occurred preferentially in slow-firing units, which were atypical of awake conditions. Consistent with no postsynaptic DA receptors on SNr neurons, iontophoretic DA was ineffective in altering discharge rates in awake conditions, but often induced weak excitations during anesthesia. Although SNr neurons are autoactive, generating discharges without any excitatory input (i.e., in vitro), their impulse activity and responses to natural neurochemical inputs are strongly affected by general anesthesia. Some alterations appear to be specific to the general anesthetic used, while others probably reflect changes in the activity of afferent inputs, brain metabolism and neurotransmitter uptake that are typical to any type of general anesthesia. Therefore, an awake, freely moving animal preparation appears to be advantageous for studying impulse activity and neurochemical interactions at single-neuron level during physiologically relevant conditions.

GABAergic mechanisms in regulating the activity state of substantia nigra pars reticulata neurons

Substantia nigra reticulata is the major output structure of the basal ganglia involved in somatosensory integration and organization of movement. While previous work in vitro and in anesthetized animal preparations suggests that these neurons are autoactive and points to GABA as a primary input regulating their activity, single-unit recording coupled with iontophoresis was used in awake, unrestrained rats to further clarify the role of tonic and phasic GABA input in maintenance and fluctuations of substantia nigra reticulata neuronal activity under physiologically relevant conditions. In contrast to glutamate, which was virtually ineffective at stimulating substantia nigra reticulata neurons in awake rats, all substantia nigra reticulata neurons tested were inhibited by iontophoretic GABA and strongly excited by bicuculline, a GABA-A receptor blocker. The GABA-induced inhibition had short onset and offset latencies, a fading response pattern (a rapid decrease in rate followed by its relative restoration), and was independent of basal discharge rate. The bicuculline-induced excitation was inversely related to discharge rate and current (dose)-dependent in individual units. However, the average discharge rate during bicuculline applications at different currents increased to a similar plateau (∼60 impulses/s), which was about twice the mean basal rates. The excitatory effects of bicuculline were phasically inhibited or completely blocked by brief GABA applications and generally mimicked by gabazine, another selective GABA antagonist. These data as well as neuronal inhibitions induced by nipecotic acid, a selective GABA uptake inhibitor, suggest that substantia nigra reticulata neurons in awake, quietly resting conditions are under tonic, GABA-mediated inhibition. Therefore, because of inherent autoactivity and specifics of afferent inputs, substantia nigra reticulata neurons are very sensitive to phasic alterations in GABA input, which appears to be the primary factor determining fluctuations in their activity states under physiological conditions. While these cells are relatively insensitive to direct activation by glutamate, and resistant to a continuous increase in GABA input, they appear to be very sensitive to a diminished GABA input, which may release them from tonic inhibition and determine their functional hyperactivity.

Up-Regulation in Expression of Vesicular Glutamate Transporter 3 in Substantia Nigra but Not in Striatum of 6-Hydroxydopamine-Lesioned Rats

Overactivity of the glutamatergic system is suggested to be closely related to the onset and pathogenesis of Parkinson’s disease. Vesicular glutamate transporters (VGLUT1, T2 and T3) are a group of glutamate transporters in neurons that are responsible for transporting glutamate into synaptic vesicles and they are key elements for homeostasis of glutamate neurotransmission. The present study was aimed to investigate the expression of VGLUT1, T2 and T3 proteins after the onset of Parkinson’s disease. A rat model of Parkinson’s disease, the 6-hydroxydopamine-lesioned rat, was employed. Immunocytochemistry revealed that VGLUT1, T2 and T3 immunoreactivity was not modulated in the striatum of the lesioned rat. Western blotting analyses also showed that there was no change in the expression of T1, T2 and T3 proteins in the striatum. In contrast, no VGLUT1 protein was detected in the substantia nigra. After the lesion, levels of VGLUT2 immunoreactivity and protein were not modulated. Significant increase of VGLUT3 immunoreactivity was observed in the perikarya of GABAergic substantia nigra pars reticulata neurons (+14.7%) although VGLUT3 protein was not modulated in the nigral tissues. VGLUT3 in GABAergic neurons is suggested to play a role in GABA synthesis. The present results may therefore implicate that VGLUT1 and T2 are not modulated in the striatum and the substantia nigra of the 6-hydroxydopamine-lesioned rat and only VGLUT3 plays a role in pathogenesis of Parkinson’s disease.

Activity of substantia nigra pars reticulata neurons during smooth pursuit eye movements in monkeys

Nuclei within the basal ganglia (BG), in particular the substantia nigra pars reticulata (SNr), subthalamic nucleus (STN) and caudate nucleus, are known to be involved in the generation of rapid or saccadic eye movements. Neurons in the SNr are active tonically and generally show a pause, but also increase, in discharge rate, for the appearance of visual stimuli and the generation of saccades. Recent experimental results in oculomotor regions of the brainstem reveal overlap in the neuronal pathways used for saccades and smooth pursuit, or slow tracking, eye movements. Whether the overlap of processing for saccades and pursuit extends to the oculomotor BG is unknown. In the present report, we were interested in whether the overlap between the pursuit and saccadic systems extends into the oculomotor BG. Using single-neuron recording and electrical stimulation techniques, we tested whether neurons within the saccade portion of the BG, the SNr, could be involved in smooth pursuit eye movements. Monkeys were required to follow visual targets with either a smooth eye movement or a saccade while we recorded from SNr neurons. We report here on SNr neuronal activity that was modulated during the performance of visually guided saccades and also during the initiation and the maintenance of smooth pursuit eye movements. Importantly, the modulation of neuronal activity during pursuit was present even when catch-up saccades were absent. The majority of SNr neurons was active tonically and their discharge ceased during pursuit, although some neurons also increased their discharge rate during smooth pursuit, similar to the behaviour reported for saccades. We also found that electrical stimulation of the SNr during the initiation of pursuit suppressed ipsiversive and, in some cases, enhanced contraversive pursuit. Our combined recording and stimulation results are consistent with the hypothesis that the overlap between the pursuit and saccadic systems extends, at least somewhat, into the BG and that the signal conveyed by the SNr can be used by the pursuit system. Like the signal for saccades, the SNr may provide a permissive disinhibition for pursuit eye movements. We hypothesize that alterations in this signal in BG diseased states such as Parkinson's may explain in part the deficits observed in smooth pursuit eye movements of these patients.

Multisensory responses and receptive field properties of neurons in the substantia nigra and in the caudate nucleus

The basal ganglia are widely regarded as structures involved in sensorimotor co-ordination, but little is known about the sensory background of their function. We publish here descriptions of the excitatory sensory responses and receptive field properties of the visual, auditory, somatosensory and multisensory caudate nucleus and substantia nigra pars reticulata neurons. Altogether 111 caudate nucleus and 124 substantia nigra sensory neurons were recorded in halothane-anaesthetized, immobilized, artificially ventilated cats. The sensory properties of the caudate and nigral neurons were found to be quite similar. A majority of the units were unimodal while a significant proportion of them were multisensory. The visual and the somatosensory modalities predominated for both nuclei. The sensory receptive fields were extremely large. The visual and auditory receptive fields covered the whole physically approachable sensory field and the somatosensory receptive fields covered the whole body surface of the animal. The receptive field properties of the multisensory caudate and nigral units were similar to those of the unimodal neurons. We observed no signs of retinotopical or somatotopical organization within the basal ganglia. The particular sensory receptive field properties, together with the significant number of multisensory neurons in the basal ganglia, suggest the existence of a multisensory pathway of presumably tectal origin involving the caudate nucleus and the substantia nigra that may serve for the sensory feedback of motor actions co-ordinated by the basal ganglia.

Two pathways for the activation of small-conductance potassium channels in neurons of substantia nigra pars reticulata

Neurons in substantia nigra pars reticulata express the messenger RNA for SK2 but not for SK3 subunits that form small-conductance, Ca 2+-dependent K + channels in dopamine neurons. To determine pathways for the activation of small-conductance, Ca 2+-dependent K + channels in substantia nigra pars reticulata neurons of rats and mice, we studied effects of the selective blocker of small-conductance, Ca 2+-dependent K + channels, apamin (0.01 or 0.3μM). Apamin diminished the afterhyperpolarization following each action potential and induced burst discharges in substantia nigra pars reticulata neurons. Apamin had a robust effect already at a low (10nM) concentration consistent with the expression of the SK2 subunit. Afterhyperpolarizations were also reduced by the Ca 2+ channel blockers Ni 2+ (100μM) and ω-conotoxin GVIA (1μM). Depletion of intracellular Ca 2+ stores did not change the afterhyperpolarization. However, we observed outward current pulses that occurred independently from action potentials and were abrogated by apamin. Apart from a faster time course, they shared all properties with spontaneous hyperpolarizations or outward currents that ryanodine receptor-mediated Ca 2+ release from intracellular stores induces in juvenile dopamine neurons. Sensitization of ryanodine receptors by caffeine silenced substantia nigra pars reticulata neurons. This effect was abolished by the depletion of intracellular Ca 2+ stores. We conclude that SK2 channels in substantia nigra pars reticulata neurons are activated by Ca 2+ influx through at least two types of Ca 2+ channels in the membrane and by ryanodine receptor-mediated Ca 2+ release from intracellular stores. Ryanodine receptors do not amplify small-conductance, Ca 2+-dependent K + channel activation by the Ca 2+ influx during a single spike. Yet, ryanodine receptor-mediated Ca 2+ release and, thereby, an activation of small-conductance, Ca 2+-dependent K + channels by intracellular Ca 2+ are available for excitability modulation in these output neurons of the basal ganglia system.