S.T. Kitai

Glutamate decarboxylase immunoreactive neurons in rat neostriatum: their morphological types and populations.

Morphological types and populations of glutamate decarboxylase (GAD)-immunoreactive neurons in rat neostriatum (Str) were studied. Str of colchicine-treated animals contained 3 types of neurons immunoreactive for GAD. The first type, which makes up 80-84% of Str neurons, was medium in size and showed moderate intensity GAD-staining. The somatic morphology of the neurons was identical to the medium-spiny projection neuron. The second type, 3-5% of Str neurons, was small to medium in size and was intensely stained for GAD. The somata of the neurons were round or oval and contained a narrow ring of cytoplasm surrounding the nucleus, which often had nuclear invaginations. There were only a few in each section of the third type, which were large, polygonal, and intensely stained, GAD-immunoreactive neurons, including all 3 types, ranged from 85-87% of the total neuron population. The present study indicated that GABAergic neurons in the Str are not a single morphological type and that most Str projection neurons are GABAergic.

Intracellular study of rat globus pallidus neurons : membrane properties and responses to neostriatal, subthalamic and nigral stimulation

Physiological properties of globus pallidus (GP) neurons were studied intracellularly in anesthetized rats. More than 70% of the neurons exhibited continuous repetitive firing of 2-40 Hz, while others exhibited periodic burst firing or no firing. The repetitively firing neurons exhibited the following properties: spike accommodation; spike frequency adaptation; continuous firing with a frequency of about 100 Hz generated by intracellular current injections; fast anomalous rectification; ramp-shaped depolarization upon injection of depolarizing current; and post-active hyperpolarization. The burst firing neurons evoked a large depolarization with multiple spikes in response to depolarizing current, and a similar response was observed after the termination of hyperpolarizing current. The few neurons which did not fire spontaneous spikes exhibited strong spike accommodation when they were stimulated by current injections. The continuously firing neurons were antidromically activated by stimulation of the neostriatum (Str) (23 of 68), the subthalamic nucleus (STh) (55 of 75), and the substantia nigra (SN) (25 of 46). The antidromic latencies of the 3 stimulus sites were very similar (about 1 ms). None of the burst firing neurons were antidromically activated. Three non-firing neurons evoked antidromic responses only after Str stimulation. Only repetitively firing neurons evoked postsynaptic responses following stimulation of the Str and the STh. Stimulation of the Str evoked initial small EPSPs with latencies of 2-4 ms and strong, short duration IPSPs with latencies of 2-12 ms. Stimulation of the STh evoked short latency EPSPs overlapped with IPSPs. Frequently, these responses induced by Str and STh stimulation were followed by other EPSPs lasting 50-100 ms. These results indicated: (1) that the GP contains at least 3 electrophysiologically different types of neurons; (2) that GP projections to the Str, the STh, and the SN are of short latency pathways; (3) that Str stimulation evokes short latency EPSPs followed by IPSPs and late EPSPs in GP neurons; and (4) that STh stimulation evokes short latency EPSPs overlapped with short latency IPSPs and late EPSPs in GP neurons.

Intracellular study of rat substantia nigra pars reticulata neurons in an in vitro slice preparation: electrical membrane properties and response characteristics to subthalamic stimulation

The electrical membrane properties of substantia nigra pars reticulata (SNR) neurons and their postsynaptic responses to stimulation of the subthalamic nucleus (STH) were studied in an in vitro slice preparation. SNR neurons were divided into two types based on their electrical membrane properties. Type-I neurons possessed (1) spontaneous repetitive firings, (2) short-duration action potentials, (3) less prominent spike accommodations, and (4) a strong delayed rectification during membrane depolarization. Type-II neurons had (1) no spontaneous firings, (2) long-duration action potentials, (3) a prominent spike accommodation, (4) a relatively large post-active hyperpolarization, and (5) a less prominent delayed rectification. These membrane properties were very similar to those observed in substantia nigra pars compacta (SNC) neurons in slice preparations. Features common to both types of neurons include that (1) the input resistance was similar, (2) they showed an anomalous rectification during strong hyperpolarizations, and (3) they were capable of generating Ca potentials. Intracellular responses of both types of SNR neurons to STH stimulation consisted of initial short-duration monosynaptic excitatory postsynaptic potentials (EPSPs) and a short-duration inhibitory postsynaptic potential (IPSP) followed by a long-duration depolarization. The IPSP was markedly suppressed by application of bicuculline methiodide and the polarity was reversed by intracellular injection of Cl-. In the preparations obtained from internal capsule-transected rats, STH-induced EPSPs had much longer durations than those observed in the normal preparations, while the amplitude of IPSPs and succeeding small-amplitude long-duration depolarizations was small. The results indicated that SNR contains two electrophysiologically different types of neurons, and that both types of neurons receive monosynaptic EPSPs from STH and IPSPs from areas rostral to STH.

Electrophysiological properties of pedunculopontine neurons and their postsynaptic responses following stimulation of substantia nigra reticulata.

Membrane properties and postsynaptic responses to stimulation of the substantia nigra reticulata (SNr) of the neurons in rat pedunculopontine nucleus (PPN) were studied in an in vitro parasagittal slice preparation using intracellular recording techniques. Based on electrical membrane properties, PPN neurons were classified into 3 types (types I, II and II). The unique feature of the type I neuron was the low threshold calcium spike while the type II neuron had various inward and outward rectifications. The type III neuron showed no such features as those observed in type I or II neurons. Some recorded neurons were intracellularly labeled with biocytin to study their morphology, and their transmitter phenotype was investigated by immunocytochemistry for choline acetyltransferase (ChAT). The type I and III neurons were found to be non-cholinergic, but 50% of the labeled type II neurons were immunopositive for ChAT. Morphological features of type II neurons were also different from type I or III neurons. The soma of the type II neuron was almost always more than twice as large as that of type I and III neurons. Inhibitory postsynaptic potentials (IPSPs) were induced in all 3 types of PPN neurons following stimulation of SNr. SNr-induced IPSPs were usually followed by a slow depolarizing potential from which rebound spikes were triggered. These rebound excitations were found only in type I and II neurons. These data indicate that heterogeneous groups of neurons exist in the PPN in terms of morphology, transmitter phenotypes and electrical membrane properties.

Electrical membrane properties of rat subthalamic neurons in an in vitro slice preparation.

The electrical membrane properties of subthalamic (STH) neurons and their response characteristics to stimulation of the internal capsule (IC) were studied in an in vitro slice preparation. Most STH neurons recorded exhibited spontaneous repetitive firing. The input resistance of STH neurons was 146 +/- 48 M omega and showed both an anomalous and a delayed rectification when the membrane was hyperpolarized or depolarized by current injections. In neurons with the membrane potential less negative than 65 mV, depolarizing current pulses generated repetitive firing with the maximum frequency of up to 500 Hz. Two types of tetrodotoxin (TTX)-resistant cobalt-sensitive potentials, slow depolarizing potential and slow action potential, were observed in STH neurons. The slow depolarizing potential had a long duration (over 500 ms in some cases) and was able to trigger repetitive firing. The slow action potential had a duration of about 30 ms and triggered a burst of firing. The slow action potential was seen only when the neurons were hyperpolarized to more negative than 65 mV by a current injection. Electrical stimulation of IC evoked monosynaptic inhibitory postsynaptic potentials (IPSPs) in most of the neurons examined. The polarity of IPSPs was reversed in the depolarizing direction by intracellular injection of Cl-. Bath application of bicuculline markedly suppressed IPSPs and unmasked monosynaptic excitatory postsynaptic potentials (EPSPs). The EPSP was able to trigger a slow depolarization with repetitive firing or a slow action potential with burst of firing when the neuron was hyperpolarized by a continuous current injection. The results demonstrated that STH neurons in an in vitro preparation have spontaneous discharges, high input resistance, capability to generate high-frequency firing, and Ca potentials. The pattern of responses of STH neurons to synaptic inputs is dependent on their membrane potentials.

Developmental regulation of a slowly-inactivating potassium conductance in rat neostriatal neurons.

In late embryonic and early post-natal rat neostriatal neurons, the voltage-dependent potassium currents activated by depolarization are largely attributable to a rapidly inactivating A-current and a delayed rectifier current. Over the first 4 weeks of post-natal life, a third potassium current emerges in most cells. This slowly inactivating conductance is distinct from the A-current and delayed rectifier in voltage-dependence, kinetics and pharmacology. The properties of this conductance suggest that it may be of central importance to the integrative behavior of neostriatal neurons by controlling such features as first spike latency and interspike interval.

Intracellular study of rat entopeduncular nucleus neurons in an in vitro slice preparation: response to subthalamic stimulation.

Responses of rat entopeduncular nucleus (EP) neurons after stimulation of the subthalamic nucleus (STh) and the morphology of the EP neurons were studied using brain slice preparations. EP neurons were classified into two types based on their electrophysiological properties as reported previously. Of 87 EP neurons, 72 were Type I and the rest were Type II. Synaptic responses to STh stimulation were different in these two cell types. STh stimulation evoked excitatory postsynaptic potentials (EPSPs) followed by strong inhibitory postsynaptic potentials (IPSPs) in Type I neurons and EPSPs without strong IPSPs in Type II neurons. The EPSPs were considered to be monosynaptic because no large change in the latency (1.7 +/- 0.5 ms) resulted by alteration of stimulus intensity. The EPSPs were reversibly suppressed by kynurenic acid in a dose-dependent manner. Bath application of (+)-tubocurarine (10-50 microM) had no effect on EPSPs or IPSPs. Bath application of bicuculline methiodide (50-100 microM) markedly suppressed IPSPs evoked by STh stimulation and at the same time increased the amplitude and duration of EPSPs without affecting the latency. In the presence of bicuculline methiodide, EPSPs could induce plateau potentials and slow action potentials. Some type I and Type II neurons were intracellularly labeled by biocytin. Type I neurons were located throughout the EP but Type II neurons were located mainly in the dorsal portion of the EP. Medium sized somata of both Type I and Type II neurons were spine-free and fusiform or round in shape. They had 3-4 thick primary dendrites with diameters of 2-5 micron that branched into thin secondary dendrites.(ABSTRACT TRUNCATED AT 250 WORDS)

Intracellular study of rat entopeduncular nucleus neurons in an in vitro slice preparation: electrical membrane properties.

Electrical properties of rat entopeduncular nucleus (EP) neurons were studied in vitro using slice preparations. Of 108 EP neurons recorded, 104 were classified into two types based on their membrane properties. Type I neurons (n = 86) possessed: (1) a strong, time-dependent anomalous rectification that was sensitive to Cs+; (2) a weak spike adaptation; and (3) a strong rebound excitation with a low threshold Ca-spike and fast spikes. Many Type I neurons displayed spontaneous repetitive firing. Some of them generated spontaneous Ca-dependent plateau potentials with fast spikes upon application of tetraethylammonium bromide. Type II neurons (n = 18) had: (1) no apparent rectification; (2) a strong spike adaptation; and (3) a ramp-shaped repolarization, similar to the A-current, at the offset of a hyperpolarizing pulse. Features common to both types included: (1) a similar range of the input resistance; (2) capability of generating high threshold Ca-spikes; and (3) generation of postactive hyperpolarizations (i.e. Ca-activated K-conductance). The great majority (Type I) of rat EP neurons share similar electrical properties. A minority of neurons (Type II) behave differently from Type I neurons and share similar properties among themselves.

Two types of A-current differing in voltage-dependence are expressed by neurons of the rat neostriatum

Transient potassium currents of the A type are thought to be important in a number of physiological processes of excitable cells, including spike repolarization and synaptic integration. This functional diversity may reflect the contribution of distinct subtypes of A channel to cellular behavior. Using the whole-cell variant of the patch clamp technique, we have found that two types of A-current are expressed in rat neostriatal neurons, one that is similar to previous descriptions in mammals and a second that is activated at considerably more depolarized potentials.

Enkephalinergic-cholinergic interaction in the rat globus pallidus: a pre-embedding double-labeling immunocytochemistry study

The synaptic relationships between leucine-enkephalin containing axon terminals and cholinergic neurons in the rat globus pallidus were studied at both light and electron microscopic levels using a high resolution pre-embedding double-labeling immunocytochemical method. Results indicated that leucine-enkephalin terminals very rarely form monosynaptic connections with cholinergic neurons in the rat globus pallidus, suggesting that enkephalinergic neostriatal efferents probably have little monosynaptic influences on the activities of pallidal cholinergic neurons.