RESUMO
The debilitating psychomotor symptoms of Huntington's disease (HD) are linked partly to degeneration of the basal ganglia indirect pathway. At early symptomatic stages, before major cell loss, indirect pathway neurons exhibit numerous cellular and synaptic changes in HD and its models. However, the impact of these alterations on circuit activity remains poorly understood. To address this gap, optogenetic- and reporter-guided electrophysiological interrogation was used in early symptomatic male and female Q175 HD mice. D2 dopamine receptor-expressing striatal projection neurons (D2-SPNs) were hypoactive during synchronous cortical slow-wave activity, consistent with known reductions in dendritic excitability and cortical input strength. Downstream prototypic parvalbumin-expressing external globus pallidus (PV+ GPe) neurons discharged at 2-3 times their normal rate, even during periods of D2-SPN inactivity, arguing that defective striatopallidal inhibition was not the only cause of their hyperactivity. Indeed, PV+ GPe neurons also exhibited abnormally elevated autonomous firing ex vivo Optogenetic inhibition of PV+ GPe neurons in vivo partially and fully ameliorated the abnormal hypoactivity of postsynaptic subthalamic nucleus (STN) and putative PV- GPe neurons, respectively. In contrast to STN neurons whose autonomous firing is impaired in HD mice, putative PV- GPe neuron activity was unaffected ex vivo, implying that excessive inhibition was responsible for their hypoactivity in vivo Together with previous studies, these data demonstrate that (1) indirect pathway nuclei are dysregulated in Q175 mice through changes in presynaptic activity and/or intrinsic cellular and synaptic properties; and (2) prototypic PV+ GPe neuron hyperactivity and excessive target inhibition are prominent features of early HD pathophysiology.SIGNIFICANCE STATEMENT The early symptoms of Huntington's disease (HD) are linked to degenerative changes in the action-suppressing indirect pathway of the basal ganglia. Consistent with this linkage, the intrinsic properties of cells in this pathway exhibit complex alterations in HD and its models. However, the impact of these changes on activity is poorly understood. Using electrophysiological and optogenetic approaches, we demonstrate that the indirect pathway is highly dysregulated in early symptomatic HD mice through changes in upstream activity and/or intrinsic properties. Furthermore, we reveal that hyperactivity of external globus pallidus neurons and excessive inhibition of their targets are key features of early HD pathophysiology. Together, these findings could help to inform the development and targeting of viral-based, gene therapeutic approaches for HD.
Assuntos
Doença de Huntington , Núcleo Subtalâmico , Animais , Gânglios da Base , Feminino , Globo Pálido/fisiologia , Doença de Huntington/metabolismo , Masculino , Camundongos , OptogenéticaRESUMO
KEY POINTS: Reciprocally connected GABAergic external globus pallidus (GPe) and glutamatergic subthalamic nucleus (STN) neurons form a key network within the basal ganglia. In Parkinson's disease and its models, abnormal rates and patterns of GPe-STN network activity are linked to motor dysfunction. Using cell class-specific optogenetic identification and inhibition during cortical slow-wave activity and activation, we report that, in dopamine-depleted mice, (1) D2 dopamine receptor expressing striatal projection neurons (D2-SPNs) discharge at higher rates, especially during cortical activation, (2) prototypic parvalbumin-expressing GPe neurons are excessively patterned by D2-SPNs even though their autonomous activity is upregulated, (3) despite being disinhibited, STN neurons are not hyperactive, and (4) STN activity opposes striatopallidal patterning. These data argue that in parkinsonian mice abnormal, temporally offset prototypic GPe and STN neuron firing results in part from increased striatopallidal transmission and that compensatory plasticity limits STN hyperactivity and cortical entrainment. ABSTRACT: Reciprocally connected GABAergic external globus pallidus (GPe) and glutamatergic subthalamic nucleus (STN) neurons form a key, centrally positioned network within the basal ganglia. In Parkinson's disease and its models, abnormal rates and patterns of GPe-STN network activity are linked to motor dysfunction. Following the loss of dopamine, the activities of GPe and STN neurons become more temporally offset and strongly correlated with cortical oscillations below 40 Hz. Previous studies utilized cortical slow-wave activity and/or cortical activation (ACT) under anaesthesia to probe the mechanisms underlying the normal and pathological patterning of basal ganglia activity. Here, we combined this approach with in vivo optogenetic inhibition to identify and interrupt the activity of D2 dopamine receptor-expressing striatal projection neurons (D2-SPNs), parvalbumin-expressing prototypic GPe (PV GPe) neurons, and STN neurons. We found that, in dopamine-depleted mice, (1) the firing rate of D2-SPNs was elevated, especially during cortical ACT, (2) abnormal phasic suppression of PV GPe neuron activity was ameliorated by optogenetic inhibition of coincident D2-SPN activity, (3) autonomous PV GPe neuron firing ex vivo was upregulated, presumably through homeostatic mechanisms, (4) STN neurons were not hyperactive, despite being disinhibited, (5) optogenetic inhibition of the STN exacerbated abnormal GPe activity, and (6) exaggerated beta band activity was not present in the cortex or GPe-STN network. Together with recent studies, these data suggest that in dopamine-depleted mice abnormally correlated and temporally offset PV GPe and STN neuron activity is generated in part by elevated striatopallidal transmission, while compensatory plasticity prevents STN hyperactivity and limits cortical entrainment.
Assuntos
Globo Pálido , Núcleo Subtalâmico , Animais , Gânglios da Base , Dopamina , Camundongos , Vias Neurais , NeurôniosRESUMO
The development of methodology to identify specific cell populations and circuits within the basal ganglia is rapidly transforming our ability to understand the function of this complex circuit. This mini-symposium highlights recent advances in delineating the organization and function of neural circuits in the external segment of the globus pallidus (GPe). Although long considered a homogeneous structure in the motor-suppressing "indirect-pathway," the GPe consists of a number of distinct cell types and anatomical subdomains that contribute differentially to both motor and nonmotor features of behavior. Here, we integrate recent studies using techniques, such as viral tracing, transgenic mice, electrophysiology, and behavioral approaches, to create a revised framework for understanding how the GPe relates to behavior in both health and disease.
Assuntos
Gânglios da Base/fisiologia , Corpo Estriado/fisiologia , Globo Pálido/fisiologia , Doença de Parkinson/fisiopatologia , Animais , Globo Pálido/anatomia & histologia , Humanos , Vias Neurais/fisiologia , Neuroglia/fisiologia , Neurônios/fisiologiaRESUMO
The frequency and pattern of activity in the reciprocally connected GABAergic external globus pallidus (GPe) and glutamatergic subthalamic nucleus (STN) are closely related to motor function. Although phasic, unitary GPe-STN inputs powerfully pattern STN activity ex vivo, correlated GPe-STN activity is not normally observed in vivo. To test the hypothesis that the GPe's influence is constrained by short-term synaptic depression, unitary GPe-STN inputs were stimulated in rat and mouse brain slices at rates and in patterns that mimicked GPe activity in vivo. Together with connectivity estimates these data were then used to simulate GPe-STN transmission. Unitary GPe-STN synaptic connections initially generated large conductances and transmitted reliably. However, the amplitude and reliability of transmission declined rapidly (τ = 0.6 ± 0.5 s) to <10% of their initial values when connections were stimulated at the mean rate of GPe activity in vivo (33 Hz). Recovery from depression (τ = 17.3 ± 18.9 s) was also longer than pauses in tonic GPe activity in vivo. Depression was the result of the limited supply of release-ready vesicles and was in sharp contrast to Calyx of Held transmission, which exhibited 100% reliability. Injection of simulated GPe-STN conductances revealed that synaptic depression caused tonic, nonsynchronized GPe-STN activity to disrupt rather than abolish autonomous STN activity. Furthermore, synchronous inhibition of tonically active GPe-STN neurons or phasic activity of GPe-STN neurons reliably patterned STN activity through disinhibition and inhibition, respectively. Together, these data argue that the frequency and pattern of GPe activity profoundly influence its transmission to the STN.
Assuntos
Globo Pálido/fisiologia , Núcleo Subtalâmico/fisiologia , Transmissão Sináptica/fisiologia , Animais , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Técnicas de Cultura de Órgãos , Distribuição Aleatória , Ratos , Ratos Sprague-Dawley , Fatores de TempoRESUMO
The subthalamic nucleus (STN) is traditionally thought to restrict movement. Lesion or prolonged STN inhibition increases movement vigor and propensity, while optogenetic excitation has opposing effects. However, STN neurons often exhibit movement-related increases in firing. To address this paradox, STN activity was recorded and manipulated in head-fixed mice at rest and during self-initiated and self-paced treadmill locomotion. We found that (1) most STN neurons (type 1) exhibit locomotion-dependent increases in activity, with half firing preferentially during the propulsive phase of the contralateral locomotor cycle; (2) a minority of STN neurons exhibit dips in activity or are uncorrelated with movement; (3) brief optogenetic inhibition of the lateral STN (where type 1 neurons are concentrated) slows and prematurely terminates locomotion; and (4) in Q175 Huntington's disease mice, abnormally brief, low-velocity locomotion is associated with type 1 hypoactivity. Together, these data argue that movement-related increases in STN activity contribute to optimal locomotor performance.
Assuntos
Locomoção , Núcleo Subtalâmico , Animais , Núcleo Subtalâmico/fisiologia , Locomoção/fisiologia , Camundongos , Masculino , Neurônios/fisiologia , Doença de Huntington/fisiopatologia , Optogenética , Camundongos Endogâmicos C57BL , MovimentoRESUMO
The subthalamic nucleus (STN) is traditionally thought to restrict movement. Lesion or prolonged STN inhibition increases movement vigor and propensity, while ontogenetic excitation typically has opposing effects. Subthalamic and motor activity are also inversely correlated in movement disorders. However, most STN neurons exhibit movement-related increases in firing. To address this paradox, STN activity was recorded and manipulated in head-fixed mice at rest and during self-initiated treadmill locomotion. The majority of STN neurons (type 1) exhibited locomotion-dependent increases in activity, with half encoding the locomotor cycle. A minority of neurons exhibited dips in activity or were uncorrelated with movement. Brief optogenetic inhibition of the dorsolateral STN (where type 1 neurons are concentrated) slowed and prematurely terminated locomotion. In Q175 Huntington's disease mice abnormally brief, low-velocity locomotion was specifically associated with type 1 hyperactivity. Together these data argue that movement-related increases in STN activity contribute to optimal locomotor performance.
RESUMO
The symptoms of Parkinson's disease (PD) are related to changes in the frequency and pattern of activity in the reciprocally connected GABAergic external globus pallidus (GPe) and glutamatergic subthalamic nucleus (STN). In idiopathic and experimental PD, the GPe and STN exhibit hypoactivity and hyperactivity, respectively, and abnormal synchronous rhythmic burst firing. Following lesion of midbrain dopamine neurons, abnormal STN activity emerges slowly and intensifies gradually until it stabilizes after 2-3 weeks. Alterations in cellular/network properties may therefore underlie the expression of abnormal firing. Because the GPe powerfully regulates the frequency, pattern, and synchronization of STN activity, electrophysiological, molecular, and anatomical measures of GPe-STN transmission were compared in the STN of control and 6-hydroxydopamine-lesioned rats and mice. Following dopamine depletion: (1) the frequency (but not the amplitude) of mIPSCs increased by â¼70%; (2) the amplitude of evoked IPSCs and isoguvacine-evoked current increased by â¼60% and â¼70%, respectively; (3) mRNA encoding α1, ß2, and γ2 GABA(A) receptor subunits increased by 15-30%; (4) the density of postsynaptic gephyrin and γ2 subunit coimmunoreactive structures increased by â¼40%, whereas the density of vesicular GABA transporter and bassoon coimmunoreactive axon terminals was unchanged; and (5) the number of ultrastructurally defined synapses per GPe-STN axon terminal doubled with no alteration in terminal/synapse size or target preference. Thus, loss of dopamine leads, through an increase in the number of synaptic connections per GPe-STN axon terminal, to substantial strengthening of the GPe-STN pathway. This adaptation may oppose hyperactivity but could also contribute to abnormal firing patterns in the parkinsonian STN.
Assuntos
Dopamina/fisiologia , Neurônios Dopaminérgicos/patologia , Globo Pálido/patologia , Transtornos Parkinsonianos/patologia , Núcleo Subtalâmico/patologia , Animais , Proteínas de Transporte/biossíntese , Proteínas de Transporte/genética , Neurônios Dopaminérgicos/fisiologia , Comportamento Exploratório/efeitos dos fármacos , Comportamento Exploratório/fisiologia , Globo Pálido/fisiopatologia , Potenciais Pós-Sinápticos Inibidores/efeitos dos fármacos , Potenciais Pós-Sinápticos Inibidores/fisiologia , Ácidos Isonicotínicos/farmacologia , Masculino , Proteínas de Membrana/biossíntese , Proteínas de Membrana/genética , Camundongos , Camundongos Endogâmicos C57BL , Degeneração Neural , Proteínas do Tecido Nervoso/análise , Proteínas do Tecido Nervoso/biossíntese , Proteínas do Tecido Nervoso/genética , Oxidopamina/toxicidade , Transtornos Parkinsonianos/fisiopatologia , Técnicas de Patch-Clamp , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Ratos , Ratos Sprague-Dawley , Receptores de GABA-A/biossíntese , Receptores de GABA-A/genética , Núcleo Subtalâmico/fisiopatologia , Transmissão Sináptica , Proteínas Vesiculares de Transporte de Aminoácidos Inibidores/biossíntese , Proteínas Vesiculares de Transporte de Aminoácidos Inibidores/genéticaRESUMO
The activity patterns of subthalamic nucleus (STN) neurons are intimately linked to motor function and dysfunction and arise through the complex interaction of intrinsic properties and inhibitory and excitatory synaptic inputs. In many neurons, hyperpolarization-activated cyclic nucleotide-gated (HCN) channels play key roles in intrinsic excitability and synaptic integration both under normal conditions and in disease states. However, in STN neurons, which strongly express HCN channels, their roles remain relatively obscure. To address this deficit, complementary molecular and cellular electrophysiological, imaging, and computational approaches were applied to the rat STN. Molecular profiling demonstrated that individual STN neurons express mRNA encoding several HCN subunits, with HCN2 and 3 being the most abundant. Light and electron microscopic analysis showed that HCN2 subunits are strongly expressed and distributed throughout the somatodendritic plasma membrane. Voltage-, current-, and dynamic-clamp analysis, two-photon Ca(2+) imaging, and computational modeling revealed that HCN channels are activated by GABA(A) receptor-mediated inputs and thus limit synaptic hyperpolarization and deinactivation of low-voltage-activated Ca(2+) channels. Although HCN channels also limited the temporal summation of EPSPs, generated through two-photon uncaging of glutamate, this action was largely shunted by GABAergic inhibition that was necessary for HCN channel activation. Together the data demonstrate that HCN channels in STN neurons selectively counteract GABA(A) receptor-mediated inhibition arising from the globus pallidus and thus promote single-spike activity rather than rebound burst firing.
Assuntos
Dendritos/fisiologia , Potenciais Pós-Sinápticos Excitadores/fisiologia , Canais Iônicos/fisiologia , Inibição Neural/fisiologia , Neurônios/fisiologia , Núcleo Subtalâmico/fisiologia , Transmissão Sináptica/fisiologia , Animais , Dendritos/efeitos dos fármacos , Dendritos/ultraestrutura , Potenciais Pós-Sinápticos Excitadores/efeitos dos fármacos , Canais Disparados por Nucleotídeos Cíclicos Ativados por Hiperpolarização , Canais Iônicos/antagonistas & inibidores , Canais Iônicos/biossíntese , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Inibição Neural/efeitos dos fármacos , Neurônios/efeitos dos fármacos , Neurônios/ultraestrutura , Canais de Potássio , Pirimidinas/farmacologia , Ratos , Ratos Sprague-Dawley , Ratos Wistar , Núcleo Subtalâmico/efeitos dos fármacos , Núcleo Subtalâmico/ultraestrutura , Transmissão Sináptica/efeitos dos fármacosRESUMO
The basal ganglia regulate our behavior through the promotion and suppression of the actions that we perform. A new study has revealed a basal ganglia feedback circuit between the striatum and globus pallidus that can powerfully inhibit locomotion in mice.
Assuntos
Gânglios da Base , Globo Pálido , Animais , Corpo Estriado , Retroalimentação , Locomoção , CamundongosRESUMO
Burst firing of substantia nigra dopamine (SN DA) neurons is believed to represent an important teaching signal that instructs synaptic plasticity and associative learning. However, the mechanisms through which synaptic excitation overcomes the limiting effects of somatic Ca(2+)-dependent K(+) current to generate burst firing are controversial. Modeling studies suggest that synaptic excitation sufficiently amplifies oscillatory dendritic Ca(2+) and Na(+) channel currents to lead to the initiation of high-frequency firing in SN DA neuron dendrites. To test this model, visually guided compartment-specific patch-clamp recording and ion channel manipulation were applied to rodent SN DA neurons in vitro. As suggested previously, the axon of SN DA neurons was typically found to originate from a large-diameter dendrite that was proximal to the soma. However, in contrast to the predictions of the model, (1) somatic current injection generated firing that was similar in frequency and form to burst firing in vivo, (2) the efficacy of glutamatergic excitation was inversely related to the distance of excitation from the axon, (3) pharmacological blockade or genetic deletion of Ca(2+) channels did not prevent high-frequency firing, (4) action potential bursts were invariably detected first at sites that were proximal to the axon, and (5) pharmacological blockade of Na(+) channels in the vicinity of the axon/soma but not dendritic excitation impaired burst firing. Together, these data suggest that SN DA neurons integrate their synaptic input in a more conventional manner than was hypothesized previously.
Assuntos
Potenciais de Ação/fisiologia , Dopamina/metabolismo , Neurônios/fisiologia , Substância Negra/fisiologia , Animais , Axônios/fisiologia , Canais de Cálcio Tipo L/genética , Canais de Cálcio Tipo L/metabolismo , Dendritos/fisiologia , Ácido Glutâmico/metabolismo , Técnicas In Vitro , Camundongos , Camundongos Knockout , Camundongos Transgênicos , Modelos Neurológicos , Neurônios/citologia , Técnicas de Patch-Clamp , Ratos , Ratos Sprague-Dawley , Canais de Sódio/metabolismo , Substância Negra/citologiaRESUMO
Substantia nigra dopamine neurons have been implicated in the initiation and invigoration of movement, presumably through their modulation of striatal projection neuron (SPN) activity. However, the impact of native dopaminergic transmission on SPN excitability has not been directly demonstrated. Using perforated patch-clamp recording, we found that optogenetic stimulation of nigrostriatal dopamine axons rapidly and persistently elevated the excitability of D1 receptor-expressing SPNs (D1-SPNs). The evoked firing of D1-SPNs increased within hundreds of milliseconds of stimulation and remained elevated for ≥ 10 min. Consistent with the negative modulation of depolarization- and Ca2+-activated K+ currents, dopaminergic transmission accelerated subthreshold depolarization in response to current injection, reduced the latency to fire, and transiently diminished action potential afterhyperpolarization. Persistent modulation was protein kinase A dependent and associated with a reduction in action potential threshold. Together, these data demonstrate that dopaminergic transmission potently increases D1-SPN excitability with a time course that could support subsecond and sustained behavioral control.
Assuntos
Dopamina/fisiologia , Neostriado/fisiologia , Neurônios/fisiologia , Neurotransmissores/fisiologia , Receptores de Dopamina D1/fisiologia , Transmissão Sináptica/fisiologia , Animais , Fenômenos Eletrofisiológicos , Feminino , Masculino , Camundongos , Neostriado/citologia , Neostriado/metabolismo , Optogenética , Técnicas de Patch-ClampRESUMO
Abnormal subthalamic nucleus (STN) activity is linked to impaired movement in Parkinson's disease (PD). The autonomous firing of STN neurons, which contributes to their tonic excitation of the extrastriatal basal ganglia and shapes their integration of synaptic input, is downregulated in PD models. Using electrophysiological, chemogenetic, genetic, and optical approaches, we find that chemogenetic activation of indirect pathway striatopallidal neurons downregulates intrinsic STN activity in normal mice but this effect is occluded in Parkinsonian mice. Loss of autonomous spiking in PD mice is prevented by STN N-methyl-D-aspartate receptor (NMDAR) knockdown and reversed by reactive oxygen species breakdown or KATP channel inhibition. Chemogenetic activation of hM3D(Gq) in STN neurons in Parkinsonian mice rescues their intrinsic activity, modifies their synaptic integration, and ameliorates motor dysfunction. Together these data argue that in PD mice increased indirect pathway activity leads to disinhibition of the STN, which triggers maladaptive NMDAR-dependent downregulation of autonomous firing.
Assuntos
Neurônios Dopaminérgicos/patologia , Regulação para Baixo , Mesencéfalo/patologia , Núcleo Subtalâmico/patologia , Animais , Neurônios Dopaminérgicos/efeitos dos fármacos , Regulação para Baixo/efeitos dos fármacos , Peróxido de Hidrogênio/toxicidade , Ativação do Canal Iônico/efeitos dos fármacos , Canais KATP/metabolismo , Masculino , Mesencéfalo/efeitos dos fármacos , Mesencéfalo/fisiopatologia , Camundongos Endogâmicos C57BL , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Atividade Motora/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Oxidopamina , Doença de Parkinson/patologia , Doença de Parkinson/fisiopatologia , Receptores de N-Metil-D-Aspartato/metabolismo , Núcleo Subtalâmico/efeitos dos fármacos , Núcleo Subtalâmico/fisiopatologiaRESUMO
The most effective treatment for late-stage Parkinson's disease is to electrically stimulate the subthalamic nucleus (STN) at high frequencies. Why this strategy works is unclear. The work by Do and Bean shows that the Na channels in STN neurons have distinctive features--like resurgence--that regulate their spiking behavior, providing new insights into the mechanism of DBS.
Assuntos
Neurônios/fisiologia , Canais de Sódio/fisiologia , Núcleo Subtalâmico/fisiologia , Animais , Estimulação Elétrica , Humanos , Doença de Parkinson/fisiopatologia , Doença de Parkinson/terapiaRESUMO
Reciprocally connected glutamatergic subthalamic nucleus (STN) and GABAergic external globus pallidus (GP) neurons normally exhibit weakly correlated, irregular activity but following the depletion of dopamine in Parkinson's disease they express more highly correlated, rhythmic bursting activity. Patch clamp recording was used to test the hypothesis that dopaminergic modulation reduces the capability of GABAergic inputs to pattern 'pathological' activity in STN neurons. Electrically evoked GABA(A) receptor-mediated IPSCs exhibited activity-dependent plasticity in STN neurons, i.e. IPSCs evoked at frequencies between 1 and 50 Hz exhibited depression that increased with the frequency of activity. Dopamine, the D(2)-like dopamine receptor agonist quinpirole and external media containing a low [Ca(2+)] reduced both the magnitude of IPSCs evoked at 1-50 Hz and synaptic depression at 10-50 Hz. Dopamine/quinpirole also reduced the frequency but not the amplitude of miniature IPSCs recorded in the presence of tetrodotoxin. D(1)-like and D(4) agonists were ineffective and D(2/3) but not D4 receptor antagonists reversed the effects of dopamine or quinpirole. Together these data suggest that presynaptic D(2/3) dopamine receptors modulate the short-term dynamics of GABAergic transmission in the STN by lowering the initial probability of transmitter release. Simulated GABA(A) receptor-mediated synaptic conductances representative of control or modulated transmission were then generated in STN neurons using the dynamic clamp technique. Dopamine-modulated transmission was less effective at resetting autonomous activity or generating rebound burst firing than control transmission. The data therefore support the conclusion that dopamine acting at presynaptic D(2)-like receptors reduces the propensity for GABAergic transmission to generate correlated, bursting activity in STN neurons.
Assuntos
Potenciais de Ação/fisiologia , Dopamina/metabolismo , Inibição Neural/fisiologia , Neurônios/fisiologia , Receptores de Dopamina D2/metabolismo , Núcleo Subtalâmico/fisiologia , Transmissão Sináptica/fisiologia , Animais , Relógios Biológicos/fisiologia , Modelos Neurológicos , Plasticidade Neuronal/fisiologia , Ratos , Ratos Sprague-Dawley , Ratos WistarRESUMO
The autonomous firing pattern of neurons in the rat subthalamic nucleus (STN) is shaped by action potential afterhyperpolarization currents. One of these is an apamin-sensitive calcium-dependent potassium current (SK). The duration of SK current is usually considered to be limited by the clearance of calcium from the vicinity of the channel. When the cell is driven to fire faster, calcium is expected to accumulate, and this is expected to result in accumulation of calcium-dependent AHP current. We measured the time course of calcium transients in the soma and proximal dendrites of STN neurons during spontaneous firing and their accumulation during driven firing. We compared these to the time course and accumulation of AHP currents using whole-cell and perforated patch recordings. During spontaneous firing, a rise in free cytoplasmic calcium was seen after each action potential, and decayed with a time constant of about 200 ms in the soma, and 80 ms in the dendrites. At rates higher than 10 Hz, calcium transients accumulated as predicted. In addition, there was a slow calcium transient not predicted by summation of action potentials that became more pronounced at high firing frequency. Spike AHP currents were measured in voltage clamp as tail currents after 2 ms voltage pulses that triggered action currents. Apamin-sensitive AHP (SK) current was measured by subtraction of tail currents obtained before and after treatment with apamin. SK current peaked between 10 and 15 ms after an action potential, had a decay time constant of about 30 ms, and showed no accumulation. At frequencies between 5 and 200 spikes s(-1), the maximal SK current remained the same as that evoked by a single action potential. AHP current did not have time to decay between action potentials, so at frequencies above 50 spikes s(-1) the apamin-sensitive current was effectively constant. These results are inconsistent with the view that the decay of SK current is governed by calcium dynamics. They suggest that the calcium is present at the SK channel for a very short time after each action potential, and the current decays at a rate set by the deactivation kinetics of the SK channel. At high rates, repetitive firing was governed by a fast apamin-insensitive AHP current that did not accumulate, but rather showed depression with increases in activation frequency. A slowly accumulating AHP current, also insensitive to apamin, was extremely small at low rates but became significant with higher firing rates.
Assuntos
Potenciais de Ação/fisiologia , Apamina/farmacologia , Cálcio/metabolismo , Canais de Potássio Ativados por Cálcio de Condutância Baixa/efeitos dos fármacos , Canais de Potássio Ativados por Cálcio de Condutância Baixa/metabolismo , Núcleo Subtalâmico/metabolismo , Animais , Canais de Cálcio/metabolismo , Técnicas de Patch-Clamp , Ratos , Ratos Sprague-Dawley , Núcleo Subtalâmico/citologia , Núcleo Subtalâmico/efeitos dos fármacos , Fatores de TempoRESUMO
The activity of the subthalamic nucleus (STN) is intimately related to movement and is generated, in part, by voltage-dependent Na(+) (Na(v)) channels that drive autonomous firing. In order to determine the principles underlying the initiation and propagation of action potentials in STN neurons, 2-photon laser scanning microscopy was used to guide tight-seal whole-cell somatic and loose-seal cell-attached axonal/dendritic patch-clamp recordings and compartment-selective ion channel manipulation in rat brain slices. Action potentials were first detected in a region that corresponded most closely to the unmyelinated axon initial segment, as defined by Golgi and ankyrin G labelling. Following initiation, action potentials propagated reliably into axonal and somatodendritic compartments with conduction velocities of approximately 5 m s(-1) and approximately 0.7 m s(-1), respectively. Action potentials generated by neurons with axons truncated within or beyond the axon initial segment were not significantly different. However, axon initial segment and somatic but not dendritic or more distal axonal application of low [Na(+)] ACSF or the selective Na(v) channel blocker tetrodotoxin consistently depolarized action potential threshold. Finally, somatodendritic but not axonal application of GABA evoked large, rapid inhibitory currents in concordance with electron microscopic analyses, which revealed that the somatodendritic compartment was the principal target of putative inhibitory inputs. Together the data are consistent with the conclusions that in STN neurons the axon initial segment and soma express an excess of Na(v) channels for the generation of autonomous activity, while synaptic activation of somatodendritic GABA(A) receptors regulates the axonal initiation of action potentials.
Assuntos
Potenciais de Ação/fisiologia , Vias Autônomas/fisiologia , Condução Nervosa/fisiologia , Núcleo Subtalâmico/fisiologia , Transmissão Sináptica/fisiologia , Potenciais de Ação/efeitos dos fármacos , Animais , Anquirinas/análise , Vias Autônomas/efeitos dos fármacos , Vias Autônomas/ultraestrutura , Axônios/efeitos dos fármacos , Axônios/fisiologia , Axônios/ultraestrutura , Dendritos/efeitos dos fármacos , Dendritos/fisiologia , Dendritos/ultraestrutura , Eletrofisiologia , Antagonistas GABAérgicos/farmacologia , Globo Pálido/fisiologia , Globo Pálido/ultraestrutura , Complexo de Golgi/ultraestrutura , Técnicas In Vitro , Masculino , Microscopia Eletrônica , Técnicas de Patch-Clamp , Ácidos Fosfínicos/farmacologia , Propanolaminas/farmacologia , Ratos , Ratos Sprague-Dawley , Receptores de GABA/fisiologia , Sódio/farmacologia , Bloqueadores dos Canais de Sódio/farmacologia , Canais de Sódio/fisiologia , Núcleo Subtalâmico/ultraestrutura , Tetrodotoxina/farmacologia , Ácido gama-Aminobutírico/farmacologiaRESUMO
The motor symptoms of Parkinson's disease are associated with abnormal, correlated, low frequency, rhythmic burst activity in the subthalamic nucleus and connected nuclei. Research into the mechanisms controlling the pattern of subthalamic activity has intensified because therapies that manipulate the pattern of subthalamic activity, such as deep brain stimulation and levodopa administration, improve motor function in Parkinson's disease. Recent findings suggest that dopamine denervation of the striatum and extrastriatal basal ganglia profoundly alters the transmission and integration of glutamatergic cortical and GABAergic pallidal inputs to subthalamic neurons, leading to pathological activity that resonates throughout the basal ganglia and wider motor system.
Assuntos
Dopamina/deficiência , Vias Neurais/fisiopatologia , Doença de Parkinson/fisiopatologia , Núcleo Subtalâmico/fisiopatologia , Animais , Córtex Cerebral/metabolismo , Córtex Cerebral/fisiopatologia , Corpo Estriado/metabolismo , Corpo Estriado/fisiopatologia , Ácido Glutâmico/metabolismo , Humanos , Vias Neurais/metabolismo , Doença de Parkinson/metabolismo , Núcleo Subtalâmico/metabolismo , Transmissão Sináptica/fisiologia , Ácido gama-Aminobutírico/metabolismoRESUMO
The glutamatergic subthalamic nucleus (STN) is a key component of the basal ganglia, a group of subcortical brain nuclei important for voluntary movement and the site of dysfunction in Parkinson's disease. The rate and pattern of STN activity is precisely regulated by the reciprocally connected GABAergic external globus pallidus (GP(e)) and glutamatergic afferents from the cortex. Subthalamic neurons possess intrinsic membrane properties that underlie the autonomous generation of action potentials and complex forms of synaptic integration. Thus, GABA acting at GABA(A) and/or GABA(B) receptors can inhibit/reset autonomous activity by deactivating postsynaptic voltage-dependent Na(+) (Na(v)) channels and generate sufficient hyperpolarization for rebound burst firing, through the de-inactivation of postsynaptic voltage-dependent Ca(2+) (Ca(v)) and Na(v) channels. Feedback inhibition from the GP(e) can therefore paradoxically and transiently increase the efficacy of subsequent excitatory synaptic inputs, and thus enhance the response of the STN to rhythmic input from the cortex. Evidence is also provided that dopamine acting at post- and presynaptic receptors in the STN may, through actions on the integrative properties of STN neurons and activity-dependent synaptic plasticity, be critical for the patterning of STN neuronal activity in vivo. Taken together, these discoveries may be relevant for the emergence of correlated, rhythmic, burst firing in the dopamine-depleted STN of patients with PD.
Assuntos
Vias Neurais/fisiologia , Neurônios/fisiologia , Núcleo Subtalâmico/fisiologia , Ácido gama-Aminobutírico/fisiologia , Potenciais de Ação/fisiologia , Animais , Relógios Biológicos/fisiologia , Dopamina/fisiologia , Humanos , Inibição Neural/fisiologia , Transmissão Sináptica/fisiologiaRESUMO
The motor symptoms of Parkinson's disease (PD) are linked to abnormally correlated and coherent activity in the cortex and subthalamic nucleus (STN). However, in parkinsonian mice we found that cortico-STN transmission strength had diminished by 50%-75% through loss of axo-dendritic and axo-spinous synapses, was incapable of long-term potentiation, and less effectively patterned STN activity. Optogenetic, chemogenetic, genetic, and pharmacological interrogation suggested that downregulation of cortico-STN transmission in PD mice was triggered by increased striato-pallidal transmission, leading to disinhibition of the STN and increased activation of STN NMDA receptors. Knockdown of STN NMDA receptors, which also suppresses proliferation of GABAergic pallido-STN inputs in PD mice, reduced loss of cortico-STN transmission and patterning and improved motor function. Together, the data suggest that loss of dopamine triggers a maladaptive shift in the balance of synaptic excitation and inhibition in the STN, which contributes to parkinsonian activity and motor dysfunction.
Assuntos
Córtex Cerebral/fisiologia , Corpo Estriado/fisiologia , Neurônios Dopaminérgicos/fisiologia , Globo Pálido/fisiologia , Núcleo Subtalâmico/fisiologia , Animais , Comportamento Animal/efeitos dos fármacos , Comportamento Animal/fisiologia , Neurônios Dopaminérgicos/metabolismo , Técnicas de Silenciamento de Genes , Locomoção/efeitos dos fármacos , Locomoção/fisiologia , Potenciação de Longa Duração/fisiologia , Masculino , Camundongos , Camundongos Transgênicos , Inibição Neural/fisiologia , Vias Neurais/fisiologia , Oxidopamina , Doença de Parkinson/fisiopatologia , Receptores de N-Metil-D-Aspartato/genética , Receptores de N-Metil-D-Aspartato/fisiologia , Transmissão Sináptica/fisiologiaRESUMO
Reciprocally connected GABAergic neurons of the globus pallidus (GP) and glutamatergic neurons of the subthalamic nucleus (STN) are a putative generator of pathological rhythmic burst firing in Parkinson's disease (PD). Burst firing of STN neurons may be driven by rebound depolarization after barrages of GABA(A) receptor (GABA(A)R)-mediated IPSPs arising from pallidal fibers. To determine the conditions under which pallidosubthalamic transmission activates these and other postsynaptic GABARs, a parasagittal mouse brain slice preparation was developed in which pallidosubthalamic connections were preserved. Intact connectivity was first confirmed through the injection of a neuronal tracer into the GP. Voltage-clamp and gramicidin-based perforated-patch current-clamp recordings were then used to study the relative influences of GABA(A)R- and GABA(B)R-mediated pallidosubthalamic transmission on STN neurons. Spontaneous phasic, but not tonic, activation of postsynaptic GABA(A)Rs reduced the frequency and disrupted the rhythmicity of autonomous firing in STN neurons. However, postsynaptic GABA(B)Rs were only sufficiently activated to impact STN firing when pallidosubthalamic transmission was elevated or pallidal fibers were synchronously activated by electrical stimulation. In a subset of neurons, rebound burst depolarizations followed high-frequency, synchronous stimulation of pallidosubthalamic fibers. Although GABA(B)R-mediated hyperpolarization was itself sufficient to generate rebound bursts, coincident activation of postsynaptic GABA(A)Rs produced longer and more intense burst firing. These findings elucidate a novel route through which burst activity can be generated in the STN, and suggest that GABARs on STN neurons could act in a synergistic manner to generate abnormal burst activity in PD.