Biomedical subjects

S T Kitai

Publications and source records attributed to S T Kitai.

At least 73 records · Page 4Linked to original sources

PubMed1990

Amygdaloid projections to the frontal cortex and the striatum in the rat.

Projections from the basolateral nucleus of the amygdala (BLA) to the frontal cortex and the striatum were studied by using Phaseolus vulgaris-leucoagglutinin (PHA-L) anterograde tracing technique in the rat. PHA-L injections into the rostral part of the BLA resulted in a dense labeling of fibers with boutons in the dorsal bank of the rhinal fissure and in the lateral and the medial agranular cortex. PHA-L injections into the caudal part of the BLA produced a dense labeling of fibers in the medial surface of the frontal cortex. In most of the cortical regions, labeled fibers were predominantly distributed in two bands: one in the deep part of layers I and II and the other, heavier band, in layers V and VI. PHA-L injections into the rostral BLA resulted in a dense labeling of fibers with boutons in the olfactory tubercle, the rostral and caudolateral portion of the nucleus accumbens, and a large region of the caudate-putamen. The labeled area of the caudate-putamen included the rostroventral area, the central area, and the area caudal to the anterior commissure and dorsal and lateral to the globus pallidus. PHA-L injections into the caudal BLA produced fiber labeling in the most rostromedial area of the caudate-putamen facing the lateral ventricle, the medial portion of the nucleus accumbens, and the lateral septum. In the rostroventral striatum, PHA-L-labeled fibers selectively innervated the matrix compartment that contains abundant somatostatin-immunoreactive fibers. Compartmental segregation was less clear in the caudodorsolateral caudate-putamen and in the nucleus accumbens. Electron microscopy revealed that PHA-L-labeled boutons in the striatum contained abundant, small, round vesicles. These boutons formed asymmetrical synapses with dendritic spines of striatal neurons.

Afferent Pathways↗

PubMed1990

Quinolinate and kainate neurotoxicity in neostriatal cultures is potentiated by co-culturing with neocortical neurons.

It has been suggested that a disorder in the regulation of excitatory amino acids (EAA) may underlie the loss of neostriatal neurons seen in Huntington's disease. The role of neocortical afferent fibers in determining the EAA sensitivity of neostriatal neurons was assessed by comparing EAA toxicity in co-cultures of neocortex and neostriatum with that of neostriatum alone. In cultures of neostriatum alone, EAAs produced only modest neuronal losses. Kainate, which tended to be the most potent excitotoxin, produced a loss of approximately 30% of the neurons after a 5-min exposure at a 1-mM concentration. In co-cultures, the sensitivity of neostriatal neurons to EAA toxicity was dramatically enhanced; toxicity was increased about two-fold for kainate and quinolinate at millimolar concentrations and as much as 8-fold for quinolinate at micromolar concentrations. The effects of EAA co-incubation with the N-methyl-D-aspartate (NMDA) receptor antagonist, 2-amino-5-phosphonovaleric acid, suggested that the toxic actions of quinolinate, but not kainate, were mediated largely by NMDA receptors.

2-Amino-5-phosphonovalerate↗

PubMed1990

Muscarinic modulation of a transient K+ conductance in rat neostriatal neurons.

Neurons of the neostriatum are richly innervated by cholinergic neurons of intrinsic origin. Both pre- and post-synaptic muscarinic receptors mediate the effects of acetylcholine (ACh). Activation of these receptors is functionally significant, particularly in Parkinson's disease. Current-clamp studies indicate that muscarinic receptors serve to decrease the responsiveness of neostriatal neurons to excitatory inputs. Here we present evidence that this effect is caused, in part, by the muscarinic modulation of the A-current, a transient outward potassium current. The voltage dependence of this current suggests that normally it enhances spike repolarization and slows discharge rate, but does not affect 'synaptic integration'. We find that under the influence of muscarinic agonists, the voltage dependence of A-current activation and inactivation is shifted towards more negative membrane potentials and the peak conductance is increased. Therefore, at relatively hyperpolarized resting potentials, ACh transiently alters the functional role of the A-current, allowing it to suppress excitatory inputs and further slow the discharge rate. But at relatively depolarized resting potentials, ACh increases excitability by removing the A-current through inactivation.

Acetylcholine↗

PubMed1990

M1 muscarinic acetylcholine receptor in cultured rat neostriatum regulates phosphoinositide hydrolysis.

Muscarinic acetylcholine receptor expression and function in cultured rat neostriatal neurons were examined. All experiments were performed on intact neurons grown in vitro for 12-14 days. The muscarinic antagonist N-[3H]methylscopolamine [( 3H]NMS) binds to a single site in cultures with a KD of 89 pM and a Bmax of 187 fmol/mg of protein, or 32,000 sites/neuron. Competition studies using [3H]NMS were performed to determine what receptor subtypes were present. Nonlinear analysis of competition curves was best described with a single binding site for atropine, pirenzepine, and AF-DX 116 (11-[[2-[(diethylamino)-methyl]-1-piperidinyl]acetyl]-5,11-dihydro- 6H-pyrido[2,3-b][1,4]benzodiazepine-6-one), with Ki values of 0.6, 62, and 758 nM, respectively. These results indicate that the muscarinic receptors present in neostriatal cultures are of the M1 subtype, having high affinity for pirenzepine and low affinity for AF-DX 116. In contrast with antagonists, carbachol displaced [3H]NMS from two sites with Ki values of 6.5 and 147 microM, with the higher-affinity form predominant (83% of sites). The M1 receptor subtype was linked to phosphoinositide turnover. Carbachol stimulated the formation of phosphoinositides with an EC50 of 37 microM and was antagonized by atropine. At equimolar doses, pirenzepine was more potent than AF-DX 116 at antagonizing the response.

Animals↗

PubMed1990

Firing patterns and synaptic potentials of identified giant aspiny interneurons in the rat neostriatum.

Intracellular recordings were made in vivo from 9 giant aspiny neurons in the neostriatum of urethane-anesthetized rats. The cells were identified by intracellular staining with HRP or biocytin. The neurons exhibited morphological features typical of neostriatal cholinergic interneurons. Six of the cells were obtained from intact animals, while 3 were recorded from rats with ipsilateral hemidecortications. Giant aspiny neurons were characterized by their slow irregular but tonic (3-10/sec) spontaneous activity and long-duration action potentials. Examination of the underlying membrane potential trajectories during spontaneous firing revealed that individual action potentials were triggered from spontaneous small (1-5 mV) depolarizing potentials. These spontaneous potentials exhibited the voltage sensitivity of ordinary EPSPs. They were much less frequent during the 80-200 msec pause in tonic afferent input that follows the excitation evoked by cortical or thalamic stimulation, and were decreased in frequency in decorticate animals. Their rise times and half-widths matched those expected for unitary synaptic potentials placed proximally on the surface of the neurons. Low-intensity stimulation of neostriatal afferents produced small short-latency EPSPs that appeared to be composed of responses identical to the spontaneous depolarizing potentials. The latencies of the EPSPs evoked from the cerebral cortex and thalamus were consistent with a monosynaptic input from both structures, but the maximal size of the EPSPs was much smaller than that evoked in spiny neurons, suggesting that a smaller number of afferent inputs make synapses with each of the aspiny cells. Giant aspiny neurons exhibited much larger input resistances and longer time constants than spiny neostriatal neurons. They also exhibited relatively linear steady-state current-voltage relationship compared to spiny projection cells. Input resistances ranged from 71-105 M omega, and time constants ranged from 17.8-28.5 msec. Analysis of the charging transients in response to current pulses yielded estimates of dendritic length of approximately 1 length constant. Repetitive firing of the neurons was limited by a powerful spike afterhyperpolarization and by a strong spike frequency adaptation. The sensitivity of the giant aspiny interneuron to a relatively small number of proximal afferent synaptic contacts, its tonic firing, and its widespread dendritic and axonal fields place it in an excellent position to act as a modulator of the excitability of neostriatal projection neurons in advance of the onset of movement-related neostriatal activity.

Action Potentials↗

PubMed1989

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.

Animals↗

PubMed1989

Intracellular study of nucleus parabrachialis and nucleus tractus solitarii interconnections.

Responses of the nucleus parabrachialis (PBN) neurons to electrical stimulation of the nucleus tractus solitarii (NTS) were investigated by intracellular recording technique in anesthetized rats. Excitatory postsynaptic potentials (EPSPs) were evoked by ipsilateral NTS stimulation in 38 PBN neurons. They were considered monosynaptic because their latencies did not change with either variations in stimulus intensity or high-frequency repetitive stimulation. The latencies of EPSPs ranged from 1.2 to 6.9 ms. PBN neurons were also antidromically activated by NTS stimulation, giving a mean axonal conduction velocity of 4.6 m/s. Some of these neurons also responded with monosynaptic EPSPs to NTS stimulation. Direct stimulation of these neurons by depolarizing current pulses elicited repetitive firing with frequencies up to 350 Hz. The morphological analysis of 5 PBN neurons labeled with horseradish peroxidase (HRP) indicates that the soma were fusiform in shape, and the size varied from 163 to 783 microns 2. All neurons had 3-5 spiny primary dendrites which extended in a predominantly mediolateral direction. Axons arose from a proximal dendritic trunk, close to the soma. The results indicated that PBN is reciprocally connected with the NTS which elicits an excitatory effect on PBN neurons.

Action Potentials↗

PubMed1989

Intracellular analysis of excitatory subthalamic inputs to the pedunculopontine neurons.

Response patterns of the pedunculopontine (PPN) area neurons to electrical stimulation of the subthalamic nucleus (STH) were investigated in anesthetized rats. Intracellular recordings demonstrated that STH stimulation evoked short duration (mean value: 11.6 ms) depolarizing potentials which were identified as excitatory postsynaptic potentials (EPSPs) by intracellular current injection. These potentials were considered monosynaptic because the latency was constant in spite of changes in stimulus intensities. The conduction velocities of STH-PPN area afferents ranged from 1.2 to 8.3 m/s (mean value: 1.6 m/s). Similar results were obtained in rats with chronic unilateral coronal lesion just rostral to STH which eliminated the rostral afferents. In some neurons, EPSPs were followed by IPSPs. PPN area neurons were also antidromically activated by STH stimulation. Some of these neurons could also be antidromically activated by substantia nigra stimulation. We could differentiate PPN area projection neurons into two groups based on their conduction velocity. One group had a conduction velocity of 6.0 m/s and the second one 1.7 m/s. The morphology of 5 HRP-labeled PPN area neurons was analyzed. The somata were fusiform, polygonal and oval in shape, and the size varied from 152 to 1310 micron 2. Two to nine dendrites emerged from the somata and extended either radially or more in one direction (e.g. rostrocaudal). Axons arose, in general, from a proximal dendritic trunk close to the soma except one and branched near the soma. These data indicate a reciprocal connection between STH and PPN area and that STH exerts excitatory influence on PPN area neurons.

Action Potentials↗

PubMed1988

An N-methyl-D-aspartate receptor mediated excitatory postsynaptic potential evoked in subthalamic neurons in an in vitro slice preparation of the rat.

Subthalamic (STH) neurons with slow EPSPs mediated by an N-methyl-D-aspartate (NMDA) receptor were studied in rat brain slice preparation. When STH neurons were intracellularly recorded with KCl-filled electrodes, stimulation of the internal capsule (IC) evoked a short duration depolarization followed by a slow excitatory postsynaptic potential (EPSP) lasting 100-200 ms. The amplitude of the slow EPSP was increased when the neuron was hyperpolarized by a low intensity current injection but was blocked when it was hyperpolarized with strong current. The slow EPSP was reversibly suppressed by application of 30-50 microM DL-2-amino-5-phosphonovareric acid (APV). STH neurons also were recorded, with potassium methylsulfate filled electrodes, in the slice preparation obtained from rats that received chronic knife cuts of the IC at the level of the entopeduncular nucleus. Stimulation of the IC immediately rostral to the STH evoked a fast EPSP followed by a slow EPSP, and IPSPs were largely eliminated in this preparation. The slow EPSP was augmented in MG-free medium and suppressed by 50 microM APV. These results suggest that NMDA receptor mediating slow EPSPs may regulate activities of STH neurons.

2-Amino-5-phosphonovalerate↗

PubMed1988

Voltage-clamp analysis of a transient potassium current in rat neostriatal neurons.

Whole cell voltage-clamp recordings were made from cultured rat neostriatal neurons. Depolarizing voltage commands evoked transient and sustained outward K-currents. The transient K-current was activated by depolarizing commands beyond -50 mV; peak current was dependent upon holding potential. Bath application of 4-aminopyridine, but not inorganic calcium channel blockers (Cd, Co, Mn), attenuated the transient current. Reversal was near the K-equilibrium potential. These properties suggest that this transient K-current is similar to the A-current described in a number of other neurons.

4-Aminopyridine↗

PubMed1988

The expression of gamma-aminobutyric acid and Leu-enkephalin immunoreactivity in primary monolayer cultures of rat striatum.

Primary monolayer cultures of rat striatum were examined for gamma-aminobutyric acid (GABA) and leucine-enkephalin (L-ENK) immunoreactivity. Cultures were established on polycation-treated glass coverslips from the striata of gestational day 17 rat embryos using a serum and insulin-supplemented medium. The proportion of GABA-immunoreactive (GABA-IR) neurons increased during the first week in vitro from approximately one third to nearly one half and remained relatively constant thereafter. On the other hand, the proportion of L-ENK-IR neurons increased gradually over the culturing period, increasing from about one-fifth of the neurons initially to one-half after 3-4 weeks in vitro. The changes in the proportions of GABA- and L-ENK-IR neurons appeared to be largely a consequence of the death of non-immunoreactive neurons, not delayed expression or induction of GABA or L-ENK traits. Light microscopic analysis of somatic-proximal neuritic morphology led to a partitioning of the neuronal population into 4 groups. GABA- and L-ENK-IR groups were heterogeneous in this regard and differed only modestly.

Animals↗

PubMed1988

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.

Animals↗

PubMed1988

Relationship of the axonal and dendritic geometry of spiny projection neurons to the compartmental organization of the neostriatum.

Intracellular injection of HRP combined with immunocytochemistry for [Leu]enkephalin was used to demonstrate striatal spiny neuron dendritic and local axonal arborizations in the same section as enkephalin-rich patches (striosomes). Cobalt intensification of the first DAB reaction prior to the immunoperoxidase steps resulted in good contrast between the black reaction product in the intracellularly labeled cells and the brown staining for [Leu]enkephalin. Serial reconstructions of the labeled cells and nearby boundaries between the enkephalin-rich striosomes and enkephalin-poor matrix allowed the relationship between the arborizations of the labeled cells and these boundaries to be established. It was also possible to examine the relationship to compartmental boundaries of a second neuronal class consisting of large, pallidallike neurons whose somatodendritic morphology was outlined by immunoperoxidase-labeled terminals. We found that spiny projection neurons in both compartments have dendritic arbors and local axonal collaterals that are confined by compartmental boundaries. The termination or recurvature of dendrites at such boundaries suggests that the cellular basis of striatal compartmental organization is provided by this class of striatal neuron. On the other hand, large pallidumlike striatal neurons were found to have dendrites that extend across compartmental boundaries. These results support previous reports that striatal spiny projection neurons preserve the compartmental segregation of parallel striatal input-output systems, whereas other classes of striatal neurons may serve to provide limited integration between compartments.

Animals↗

PubMed1987

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.

Action Potentials↗

PubMed1987

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.

Animals↗

PubMed1987

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.

Animals↗

PubMed1987

Efferent projections of the subthalamic nucleus in the rat: light and electron microscopic analysis with the PHA-L method.

Efferent projections of rat subthalamic nucleus were studied by use of the axonal transport of phaseolus vulgaris-leucoagglutinin (PHA-L), and the results were analyzed with light and electron microscopes. PHA-L injections in the subthalamic nucleus (STH) resulted in heavy labeling of fiber plexus with en passant boutons and terminals in the pallidal complex, i.e., the entopeduncular nucleus (EP), the globus pallidus (GP) and the ventral pallidum (VP), and the substantia nigra pars reticulata (SNR). Labeling in GP was characterized by two distinct bands of labeled terminals oriented dorsoventrally, whereas labeling in SNR was patchy. STH efferents to the pallidum and SNR displayed a mediolateral topographic organization. With regard to dorsoventral organization, projections to GP were inverted, but those to SNR were not. There were moderate projections to the neostriatum and sparse projections to the frontal cortex, substantia innominata, substantia nigra pars compacta (SNC), pedunculopontine tegmental nucleus, ventral part of the central gray matter including the dorsal raphe nucleus, and the mesencephalic and pontine reticular formation. PHA-L injections in the zona incerta and the lateral hypothalamic area resulted in fiber and terminal labelings in many structures, including the basal forebrain, EP, SNC, and other brainstem areas that overlap with some of the terminal sites of STH projections. Ultrastructural observations of PHA-L labeled processes in GP and SNR revealed that STH terminals in both structures contained small pleomorphic vesicles and formed asymmetrical contacts. These contacts were mainly on dendritic shafts, but some were on somata. It also was observed that the myelinated axons of STH neurons lost their myelin after reaching their target areas and the synaptic boutons arose from relatively thin unmyelinated axons.

Animals↗

PubMed1986

Electrical membrane properties of rat substantia nigra compacta neurons in an in vitro slice preparation.

The electrical membrane properties of rat substantia nigra pars compacta (SNC) neurons were studied in an in vitro slice preparation. Some of the recorded neurons were intracellularly labeled with HRP and were found to have morphological characteristics resembling the presumed SNC dopaminergic neurons, as reported by others. The input resistance of SNC neurons at resting membrane potential ranged between 70 and 250 M omega. The membrane resistance showed strong anomalous rectification when the membrane was hyperpolarized by current injection. The anomalous rectification was decreased by the addition of tetraethylammonium bromide (TEA) to the bathing Ringer solution. Injection of depolarizing current or termination of hyperpolarizing current induced slow depolarizing potentials. Their amplitude was dependent on the membrane potential and the current intensity. In neurons treated with tetrodotoxin (TTX) and TEA, slow action potentials were triggered from the slow depolarizing potentials. Both the slow depolarizing potential and slow action potential were TTX resistant and abolished by superfusion of Ca2+-free medium. Long duration hyperpolarizations were observed following the injection of depolarizing current pulses. The hyperpolarization was abolished by the superfusion of Ca2+-free medium or decreased by addition of TEA to the Ringer solution indicating an involvement of a Ca2+-dependent K+-conductance in generation of the hyperpolarization. The long duration hyperpolarization was also observed following action potentials. The spike after hyperpolarization consisted of an initial short duration fast component and a long lasting component. The amplitude of both components seems to be reduced but not abolished by TEA (up to 10 mM). When hyperpolarizing current pulses were applied to neurons that were held either continuously depolarized or were superfused with Ca2+-free medium, the pattern of the membrane potential after the offset of current pulses consisted of an initial fast and a later slow ramp-shaped phase. The latter was associated with a membrane conductance increase and interpreted to be due to an early K+ current. This early K+ current was relatively resistant to TEA. Injections of strong depolarizing currents triggered action potentials with multiple inflections on their rising phase. The amplitudes of action potentials changed abruptly during current application. These data indicate that SNC neurons have multiple generation sites for action potential.

Action Potentials↗