Spiny Cells Research Articles

Synaptic mechanisms of NMDA-mediated hyperpolarization in lateral amygdaloid projection neurons.

Synaptic mechanisms underlying NMDA-mediated responses of neurons in the guinea pig lateral amygdala (AL) were investigated in in vitro slice preparations. Local application of NMDA resulted in initial hyperpolarization of pyramidal-like spiny cells (projection neurons), followed by prolonged depolarization. The slow depolarization represented a direct postsynaptic effect of NMDA, whereas the initial hyperpolarization was induced presynaptically through activation of GABAergic interneurons and was sensitive to blockade by tetrodotoxin as well as the GABA(A)-receptor antagonist bicuculline. Application of NMDA resulted in AP-5-sensitive, lasting depolarization also in putative interneurons of the AL suggesting direct activation of GABAergic interneurons by NMDA. These data indicate that interneurons in the rat lateral amygdala possess functional NMDA receptors, which may contribute to the predominantly inhibitory synaptic responses in amygdaloid neurons following activation through afferent input systems.

Dopamine and synaptic plasticity in the neostriatum

After the unilateral destruction of the dopamine input to the neostriatum there are enduring changes in rat behaviour. These have been ascribed to the loss of dopamine and the animals are often referred to as 'hemiparkinsonian'. In the denervated neostriatum, we have shown that not only are the tyrosine hydroxylase positive boutons missing, but also the medium sized densely spiny output cells have fewer spines. Spines usually have asymmetric synapses on their heads. In a recent stereological study we were able to show that there is a loss of approximately 20% of asymmetric synapses in the lesioned neostriatum by 1 mo after the lesion. Current experiments are trying to establish the specificity of this loss. So far we have evidence suggesting that there is no obvious preferential loss of synapses from either D1 or D2 receptor immunostained dendrites in the neostriatum with damaged dopamine innervation. These experiments suggest that dopamine is somehow necessary for the maintenance of corticostriatal synapses in the neostriatum. In a different series of experiments slices of cortex and neostriatum were maintained in vitro in such a way as to preserve at least some of the corticostriatal connections. In this preparation we have been able to show that cortical stimulation results in robust excitatory postsynaptic potentials (EPSPs) recorded from inside striatal neurons. Using stimulation protocols derived from the experiments on hippocampal synaptic plasticity we have shown that the usual consequence of trains of high frequency stimulation of the cortex is the depression of the size of EPSPs in the striatal cell. In agreement with similar experiments by others, the effect seems to be influenced by NMDA receptors since the unblocking of these receptors with low Mg++ concentrations in the perfusate uncovers a potentiation of the EPSPs after trains of stimulation. Dopamine applied in the perfusion fluid round the slices has no effect but pulsatile application of dopamine, close to the striatal cell being recorded from, and in temporal association with the cortical trains, leads to a similar LTP like effect. The reduction of K+ channel conductance in the bath with TEA also has the effect of making cortical trains induce potentiation of corticostriatal transmission. TEA applied only to the cell being recorded from has no similar effect; the cortical stimulation again depresses the EPSP amplitude, so the site of action of TEA may well be presynaptic to the striatal cell. The morphological and physiological experiments may not necessarily be related but it is tempting to suggest that dopamine protects some corticostriatal synapses by potentiating them but that in the absence of dopamine others simply disconnect and are no longer detectable on electron microscopy.

Addiction, dopamine, and the molecular mechanisms of memory.

We thank Pier Piazza, Eric Nestler, Charles O'Brien, Howard Eichenbaum, and Eric Kandel for their helpful comments and criticisms of earlier versions of the manuscript.

The synaptic framework for chemical signaling in nucleus accumbens.

Our knowledge of the organization of the nucleus accumbens has been greatly advanced in the last two decades, but only now are we beginning to understand the complex neural circuitry that underlies the mix of behaviors attributed to this nucleus. Superimposed on the neurochemically defined territories of the shell and core are four or more conduits for information flow. Each of these behaviorally relevant pathways can be characterized by the spatial distribution of inputs to its central unit: the GABAergic projection neuron, a spiny cell that also contains the opioid peptides, enkephalin or dynorphin. In this review, current models of accumbal circuits will be examined and, with the aid of recent anatomical findings, further extended to shed light on how functionally diverse information is processed in this nucleus. However complex, accumbal wiring is not fixed, and, as we will show, psychostimulants, dopamine-deleting lesions, and chronic blockade of dopaminergic receptors can alter the anatomical substrate, synaptology, and neurotrophic factors that govern circuits through the shell and core.

Changes in excitatory and inhibitory circuits of the rat hippocampus 12–14 months after complete forebrain ischemia

Changes in interneuron distribution and excitatory connectivity have been investigated in animals which had survived 12–14 months after complete forebrain ischemia, induced by four-vessel occlusion. Anterograde tracing with Phaseolus vulgaris leucoagglutinin revealed massive Schaffer collateral input even to those regions of the CA1 subfield where hardly any surviving pyramidal cells were found. Boutons of these Schaffer collaterals formed conventional synaptic contacts on dendritic spines and shafts, many of which likely belong to interneurons. Mossy fibres survived the ischemic challenge, however, large mossy terminals showed altered morphology, namely, the number of filopodiae on these terminals decreased significantly. The entorhinal input to the hippocampus did not show any morphological alterations. The distribution of interneurons was investigated by neurochemical markers known to label functionally distinct GABAergic cell populations. In the hilus, spiny interneurons showed a profound decrease in number. This phenomenon was not as obvious in CA3, but the spiny metabotropic glutamate receptor 1α-positive non-pyramidal cells, some of which contain calretinin or substance P receptor, disappeared from stratum lucidum of this area. In the CA1 region, somatostatin immunoreactivity disappeared from stratum oriens/lacunosum-moleculare-associated cells, while in metabotropic glutamate receptor 1α-stained sections these cells seemed unaffected in number. Other interneurons did not show an obvious decrease in number. In stratum radiatum of the CA1 subfield, some interneuron types had altered morphology: the substance P receptor-positive dendrites lost their characteristic radial orientation, and the metabotropic glutamate receptor 1α-expressing cells became extremely spiny. The loss of inhibitory interneurons at the first two stages of the trisynaptic loop coupled with a well-preserved excitatory connectivity among the subfields suggests that hyperexcitability in the surviving dentate gyrus and CA3 may persist even a year after the ischemic impact. The dorsal CA1 region is lost; nevertheless hyperactivity, if it occurs, may have a route to leave the hippocampus via the longitudinally extensive axon collaterals of CA3 pyramidal cells, which may activate the subiculum and entorhinal cortex with a relay in the surviving ventral hippocampal CA1 region.

Muscarinic receptors differentially modulate the persistent potassium current in striatal spiny projection neurons.

Cholinergic regulation of striatal spiny projection neuron activity is predominantly mediated through muscarinic receptor modulation of several subclasses of ion channels. Because of its critical role in governing the recurring episodes of hyperpolarization and depolarization characteristic of spiny neurons in vivo, the 4-aminopyridine-resistant, persistent potassium (K+) current, IKrp, would be a strategic target for modulation. The present results show that IKrp can be either suppressed or enhanced by muscarinic receptor stimulation. Biophysical analysis demonstrated that the depression of IKrp was associated with a hyperpolarizing shift in the voltage dependence of inactivation and a reduction in maximal conductance. By contrast, the enhancement of IKrp was linked to hyperpolarizing shifts in both activation and inactivation voltage dependencies. Viewed in the context of the natural activity of spiny neurons, muscarinic depression of IKrp should uniformly increase excitability in both hyperpolarized and depolarized states. In the hyperpolarized state, the reduction in maximal conductance should bolster the efficacy of impending excitatory input. Likewise, in the depolarized state, the decreased availability of IKrp produced by the shift in inactivation should enhance ongoing synaptic input. The alterations associated with enhancement of IKrp are predicted to have a more dynamic influence on spiny cell excitability. In the hyperpolarized state, the negative shift in activation should increase the flow of IKrp and attenuate subsequent excitatory synpatic input; whereas once the cell has traversed into the depolarized state, the negative shift in inactivation should reduce the availability of this current and diminish its influence on the existing excitatory barrage.

Connectivity and Convergence of Single Corticostriatal Axons

The distribution of synapses formed by corticostriatal neurons was measured to determine the average connectivity and degree of convergence of these neurons and to search for spatial inhomogeneities. Two kinds of axonal fields, focal and extended, and two striatal tissue compartments, the patch (striosome) and matrix, were analyzed separately. Electron microscopic examination revealed that both kinds of corticostriatal axons made synapses at varicosities that could be identified in the light microscope, and each varicosity made a single synapse. Thus, the distribution of varicosities was a good estimate of the spatial distribution of synapses. The distance between axonal varicosities was measured to determine the density of synaptic connections formed by one axon within the volume occupied by a striatal neuron. Intersynaptic distances were distributed exponentially, except that synapses were rarely located <4 microm apart. The mean distance between synapses was approximately 10 microm, so axons made a maximum of 40 synapses within the dendritic volume of a spiny neuron. There are approximately 2840 spiny neurons located within the volume of the dendrites of one spiny cell (Oorschot, 1996), so each axon must contact </=1.4% of all cells in its axonal arborization. Within the same volume there are approximately 30.5 million asymmetric synapses (Ingham et al., 1996), approximately half of which are cortical in origin. Thus, approximately 380,000 cortical axons innervate the volume of the dendritic tree of one spiny cell. Striatal neurons with totally overlapping dendritic volumes have few presynaptic cortical axons in common, and cortical cells with overlapping axons have few striatal target neurons in common. These results explain the absence of redundancy in the responses of neurons located near each other in the striatum.

Synaptic interactions between smooth and spiny neurones in layer 4 of cat visual cortex in vitro.

1. Dual intracellular recording was used to examine the interactions between neighbouring spiny (excitatory) and smooth (inhibitory) neurones in layer 4 of cat visual cortex in vitro. Synaptic connections were found in seventeen excitatory-inhibitory neurone pairs, along with one inhibitory-inhibitory connection. 2. Fast excitatory inputs onto smooth neurones (basket cells) from spiny cells (spiny stellate or pyramidal cells) (n = 6) produce large excitatory postsynaptic potentials (EPSPs) of up to 4 mV mean amplitude, whereas basket cells evoke slower inhibitory postsynaptic potentials (IPSPs) in their postsynaptic targets (n = 17), of smaller amplitude (up to 1.6 mV at membrane potentials of -60 mV). 3. Both types of PSP appear to be multiquantal, and both may exhibit depression of up to 60 % during short trains of presynaptic spikes. This depression can involve presynaptic and/or postsynaptic factors. 4. One-third (n = 5) of the spiny cell-smooth cell pairs tested were reciprocally connected, and in the one pair for which the suprathreshold interactions were comprehensively investigated, the pattern of basket cell firing was strongly influenced by the activity in the connected excitatory neurone. The basket cell was only effective in inhibiting spiny cell firing when the excitatory neurone was weakly driven.

Laminar Distribution of Fos/Calcium-Binding Protein and Fos/Neurofilament Protein-Labeled Neurons in Rat Motor and Sensory Cortex after Picrotoxin-Induced Seizures

Cerebrocortical Fos induction after picrotoxin-induced seizure occurs in spiny neurons and, to a lesser extent, in neurons defined by calcium-binding protein immunoreactivity. In motor and sensory cortex of rats we have defined the laminar distribution of Fos expression in these neurons. Initially we defined the laminar distributions of parvalbumin-, calbindin-D 28K-, and calretinin-immunoreactive aspiny neurons; these were unique for each class and similar across cortical regions. Spiny cells defined by SMI32 immunoreactivity were distributed with two peaks and there were differences between cortical regions. Parvalbumin-immunoreactive neurons exhibited peak numbers where numbers of SMI32-immunoreactive neurons were low. The distribution of Fos induction across laminae matched that of its class for calbindin-D 28K and calretinin neurons; however, Fos induction was less in infragranular compared with supragranular for parvalbumin in motor cortex and SMI32 containing neurons in both cortices. In both these latter cell classes Fos induction was inversely correlated with neuronal size. It is suggested that cell size within some cell classes is one factor that determines the extent of Fos induction within that class following seizures.

Long-term plasticity at excitatory synapses on aspinous interneurons in area CA1 lacks synaptic specificity.

The synaptic specificity of long-term potentiation (LTP) was examined at synapses formed on aspinous dendrites of interneurons whose somata were located in the pyramidal cell layer of hippocampal area CA1. Intracellular recordings from slices prepared from rats were used to monitor excitatory postsynaptic potentials (EPSPs) elicited by extracellular stimulation in stratum radiatum. Two synaptic inputs were evoked at 0.5 Hz by stimulating axons adjacent to stratum pyramidale and s. lacunosum-moleculare. After obtaining baseline recordings (>/=10 min), one of the EPSPs was conditioned. The protocol involved tetanic stimulation, sometimes combined with somatic depolarization. Low-frequency stimulation of the two pathways was then resumed and EPSPs were recorded for <30 min. We observed both homosynaptic and heterosynaptic changes in synaptic strength. LTP and long-term depression (LTD) were seen in both pathways and all possible combinations of changes in the two EPSPs were observed, including heterosynaptic LTP associated with either homosynaptic LTP or LTD. Intracellular 1,2-bis (2-aminophenoxy)-ethane-N,N,N',N'-tetraacetic acid (10 mM) abolished alterations in synaptic strength. When axons in s. radiatum synapse onto a spiny pyramidal cell, synaptic specificity of LTP is preserved. However the results obtained from aspinous interneurons show that synaptic specificity of LTP is lost. These results are consistent with the hypothesis that spines provide postsynaptic mechanism(s) for conferring specificity to LTP.

Increased hippocampal dendritic expression of non-phosphorylated MAP2 protein in schizophrenia

The possible co-existence of calretinin with other calcium binding proteins, parvalbumin and calbindin D28k, and with GABA, was studied in non-pyramidal cells of the rat dorsal hippocampal formation, using the mirror technique. The majority of the calretinin-containing neurons (83%) were found to be immunoreactive for GABA (79% in the dentate gyrus, 84% in the CA2–3, and 88% in the CA1 subfield). Most of the GABA-negative calretinin-immunoreactive neurons were located in the hilus of the dentate gyrus and in stratum lucidum of the CA3 subfield. Detailed analysis of the calretinin-immunoreactive cells of these subfields revealed that the two morphologically distinct types of calretinin neurons, i.e. the spiny and the spine-free cells, differ in their immunoreactivity for GABA. The overwhelming majority (92%) of the spine-free neurons were GABA-positive, whereas the immuno-reactivity of spiny cells was ambiguous. At the sensitivity threshold of the immunocytochemical techniques used in the present study, most of the spiny cells (89%) had to be considered as GABA-negative, although the staining intensity in their cell bodies was somewhat above background level. Colchicine treatment resulted in a degeneration of calretinin-immunoreactive neurons; therefore, its effect on the GABA content of spiny neurons could not be evaluated. Nevertheless, the observations suggest that calretinin-containing neurons are heterogeneous both morphologically and neurochemically.Examination of the co-existence of calcium binding proteins revealed that none of the hippocampal cells contained both calretinin and parvalbumin in any regions of the hippocampal formation. Some overlap was detected between the calretinin- and the calbindin D28k-containing cell populations, 5.1% of the former and 6.2% of the latter were immunoreactive for both calcium binding proteins. This may be due to a small degree of cross-reactivity of the calbindin D28k antiserum with calretinin.Thus, our results demonstrate that the majority of calretinin-immunoreactive neurons are GABAergic and represent a subpopulation of non-pyramidal cells with no or only a negligible overlap with the subpopulations containing the other calcium binding proteins, parvalbumin and calbindin.

Physiological properties of inhibitory interneurons in cat striate cortex.

Physiological and morphological properties of identified interneurons in the striate cortex of the cat were studied in vivo by intracellular recording and staining with biocytin. In conformity with in vitro studies, these non-pyramidal fast spiking cells have very brief action potentials associated with a high rate of fall, and a large hyperpolarizing afterpotential. These cells show high discharge rates, little or no spike frequency adaptation in response to depolarizing current injection, as well as a diverse range of firing patterns. Three of the cells were labeled and were found to be aspiny or sparsely spiny basket cells, with bitufted or radial dendritic arrangements, in layers II-IV. Their axonal arborizations were more dense near their somata and extended horizontally or vertically. Of 13 visually responsive cells tested, the receptive field properties of six cells and the orientation and direction preferences of eight cells were determined. Five of the successfully mapped cells had simple receptive fields while one had a complex receptive field type. The orientation and direction tuning properties of the overlapping set of eight cells showed a broad spectrum ranging from unselective to tightly tuned. The majority exhibited a clear preference for orientation and none of the cells were clearly direction selective. Quantitative analysis of the temporal properties of the spike trains during visual stimulation and spontaneous activity revealed that these cells do not exhibit any significant periodic activity, and fired at rates that were well below their maximum in response to depolarizing current pulses.

Massive Autaptic Self-Innervation of GABAergic Neurons in Cat Visual Cortex

Autapses are transmitter release sites made by the axon of a neuron on its own dendrites. We determined the numbers and precise subcellular position of autapses on different spiny and smooth dendritic cell types using intracellular biocytin filling in slices of adult neocortex. Potential self-innervation was light microscopically assessed on 10 pyramidal cells, 7 spiny stellate cells, and 41 smooth dendritic neurons from cortical layers II-V. Putative autapses occurred on each smooth dendritic neuron and on seven pyramids, but not on spiny stellate cells. However, electron microscopic examination of all light microscopically predicted sites on pyramids (n = 28) showed only one case of self-innervation with two autapses on dendritic spines. Interneurons were classified by postsynaptic target distribution () and all putative autapses of seven basket, three dendrite-targeting, and three double bouquet cells were scrutinized. All basket and dendrite-targeting cells established self-innervation, the number of autapses being 12 +/- 7 and 22 +/- 12 (mean +/- SD), respectively; only one of the double bouquet cells formed autapses (n = 3). Basket cell autapses (n = 74) were closer to the soma (12.2 +/- 22.3 microm) than autapses established by dendrite-targeting cells (51.8 +/- 49.9 microm; n = 66). The degree of self-innervation is cell type-specific. Unlike on spiny cells, autapses are abundant on GABAergic basket and dendrite-targeting interneurons, with subcellular location similar to that of synapses formed by the parent cell on other neurons. The extensive self-innervation may modulate integrative properties and/or the firing rhythm of the neuron in a manner temporally correlated with its own activity.

Neuronal composition and morphology in layer IV of two vibrissal barrel subfields of rat cortex.

The technique of intracellular injection in fixed, flattened slices was used to study neuronal composition and morphology in the postero-medial barrel subfield (PMBSF) and the antero-lateral barrel subfield (ALBSF) in layer IV of rat cortex. The PMBSF and the ALBSF contain the cortical representation of the mystacial and rostral snout vibrissae respectively. Neuronal composition differed between the PMBSF and the ALBSF. Modified pyramidal cells were the most numerous neuronal type in the PMBSF (73.1%), whereas spiny multipolar (stellate) neurons were the most numerous type in the ALBSF (40.9%). Tangential dendritic field areas of modified pyramidal cells and spiny multipolar cells in the barrels of the two barrel subfields were compared. Dendritic field areas of spiny multipolar neurons located in the barrels of the PMBSF and the ALBSF were similar (mean +/- SD; 2.44 +/- 1.83 x 10(4) and 2.88 +/- 1.47 x 10(4) microns2 respectively). Likewise, there was no significant difference in 'basal' dendritic field area of modified pyramidal neurons located in the barrels of the two different barrel subfields (4.63 +/- 1.96 x 10(4) and 4.45 +/- 1.81 x 10(4) microns2 for PMBSF and ALBSF respectively). The mean cross-sectional area of PMBSF barrels (20.5 +/- 5.69 x 10(4) microns2) in which neurons were injected was approximately seven times larger than that of the ALBSF (2.94 +/- 1.46 x 10(4) microns2). Thus, on average, the dendritic territories of these two neuronal classes sample a larger proportion of the cross-sectional area of the barrels in the ALBSF than in the PMBSF. We conclude that the close relationship between basal dendritic field area of supragranular pyramidal neurons and module size, reported in studies of other sensory areas, is not evident in all barrel subfields of the rat.

Distribution of inositol-1,4,5-trisphosphate and ryanodine receptors in rat neostriatum

The distribution of the inositol-1,4,5-trisphosphate (IP3) and the cardiac form of the ryanodine receptor, two intracellular calcium channels, was examined in the rat neostriatum. Both IP3 and ryanodine receptor labeling occurred within striatal medium spiny cells but only ryanodine receptor labeling was present in choline acetyltransferase- and parvalbumin-positive interneurons. IP3 receptor labeling was observed within cell bodies, dendrites and spines of spiny striatal neurons, as seen at both the light and electron microscopic levels. Subcellular labeling for the ryanodine receptor was restricted to cell bodies and proximal dendrites when a polyclonal antibody raised against a peptide sequence from the dog cardiac ryanodine receptor was employed. More extensive dendritic labeling was seen using monoclonal antibody MA3-916, also raised against the canine cardiac ryanodine receptor. At the ultrastructural level, labeled dendritic spines were observed frequently with the monoclonal but not the polyclonal antibody. Ryanodine receptor labeling also was present within astrocytic processes surrounding blood vessels and within the neuropil, regardless of the antibody used. The results of these studies suggest that the ryanodine receptor plays a general role in intracellular calcium regulation within striatal cells while the IP3 receptor plays a specialized role within spiny neurons.

μ-Opioid Receptors Are Localized to Extrasynaptic Plasma Membranes of GABAergic Neurons and Their Targets in the Rat Nucleus Accumbens

The activation of mu-opioid receptors in the nucleus accumbens (Acb) produces changes in locomotor and rewarding responses that are believed to involve neurons, including local gamma-aminobutyric acid (GABA)ergic neurons. We combined immunogold-silver detection of an antipeptide antiserum against the cloned mu-opioid receptor (MOR) and immunoperoxidase labeling of an antibody against GABA to determine the cellular basis for the proposed opioid modulation of GABAergic neurons in the rat Acb. MOR-like immunoreactivity (MOR-LI) was localized prominently to plasma membranes of neurons having morphological features of both spiny and aspiny cells, many of which contained GABA. Of 351 examples of profiles that contained MOR-LI and GABA labeling, 65% were dendrites. In these dendrites, MOR-LI was seen mainly along extrasynaptic portions of the plasma membrane apposed to unlabeled terminals and/or glial processes. Dually labeled dendrites often received convergent input from GABAergic terminals and/or from unlabeled terminals forming asymmetric excitatory-type synapses. Of all profiles that contained both MOR and GABA immunoreactivity, 28% were axon terminals. MOR-containing GABAergic terminals and terminals separately labeled for MOR or GABA formed synapses with unlabeled dendrites and also with dendrites containing MOR or GABA. Our results indicate that MOR agonists could modulate the activity of GABA neurons in the Acb via receptors located mainly at extrasynaptic sites on dendritic plasma membranes. MOR ligands also could alter the release of GABA onto target dendrites that contain GABA and/or respond to opiate stimulation.

Neostriatal cell subtypes and their functional roles

The neostriatum is considered to be involved in the induction of purposeful movements or in the suppression of other movements through the internal segment of the globus pallidus (GP; the entopeduncular nucleus in the rodents) and the substantia nigra (SN) to which the striatal spiny neurons project. To understand how the striatum fulfills these functions, it is necessary to know the physiological and morphological characteristics of its constituent neurons. Aspiny interneurons in the striatum are considered to receive various excitatory inputs and to contribute importantly to determining whether spiny projection neurons fire or not. Both spiny and aspiny striatal cells have been shown to be heterogeneous in their physiological, chemical and connection characteristics. In this article, how these cell subtypes are organized in the local circuitry of the striatum and their physiological roles in the basal ganglia are discussed.

Actions of Substance P on Rat Neostriatal NeuronsIn Vitro

Actions of substance P (SP) on the neostriatal neurons in in vitro rat slice preparations were studied via whole-cell patch-clamp recording. Almost all large aspiny neurons (cholinergic cells) and half of the low-threshold spike (LTS) cells (somatostatin/ NOS-positive cells) showed depolarization or an inward shift of the holding currents in response to bath-applied SP in a dose-dependent manner. In contrast, no responses were observed in fast-spiking (FS) cells (parvalbumin-positive cells) and medium spiny cells. Spike discharges followed by slow EPSPs/EPSCs were evoked by intrastriatal electrical stimulation in the large aspiny neurons. Pretreatment with [D-Arg1, D-Pro2, D-Trp7,9, Leu11]-SP, an antagonist of the SP receptor, reversibly suppressed the induction of the slow EPSPs/EPSCs and unmasked slow IPSCs. The SP-induced inward current, although almost unchanged even after the blockade of Ih channels and voltage-dependent Na+, Ca2+, and K+ channels, changed its amplitude according to the Na+ concentration used in both the large aspiny neurons and LTS cells. Thus, the cation current could account for virtually all of the inward current at resting levels in both neurons. These results suggest that the firing of afferent neurons such as striatonigral medium spiny neurons, one of the possible sources of SP, would increase the firing probability of the two types of interneurons of the neostriatum by SP-receptor-mediated opening of tetrodotoxin-insensitive cation channels.

Effects of Ovariectomy on GnRH Neuronal Morphology inRhesus Monkey (Macaca mulatta)

Gonadotropin releasing hormone (GnRH) neurons are typically simple, fusiform cells; however, over the course of prepubertal development increasing numbers take on a 'spiny' appearance. Following gonadectomy there is a decrease in the frequency of these spiny GnRH neurons. These observations which were made in the rat suggest that GnRH neurons are directly affected by the gonadal steroid milieu, though they do not themselves contain receptors for these steroidal hormones. In that there are important species differences in the hypothalamic-pituitary-gonadal axis between rats and primates, the present study was undertaken to determine whether a reduction in ovarian hormones would produce similar changes in the morphology of GnRH neurons in the monkey. A further aim was to determine whether such changes were localized to a specific brain region. Immunocytochemically defined GnRH neurons were compared in adult rhesus macaques which had been ovariectomized for 6 weeks to 2 years (n = 7) and intact, cycling animals (n = 8). Within the intact group, there were significantly more spiny GnRH neurons in the medial basal hypothalamus (MBH) than in the preoptic area (POA) (about 50% of the total in the MBH compared to 33% in the POA). Following ovariectomy the frequency of spiny cells in the MBH dropped to less than 30%, but was not significantly reduced in the POA. These results suggest that changes in systemic gonadal steroid levels result in changes in the morphology of GnRH neurons preferentially in the MBH, a region that is considered critical in the generation of GnRH pulsatile release in the monkey.

S-8-5 - Cholinergic and GABAergic interneurons in the striatum

In the striatum, interneurons have not been as well characterized physiologically as the spiny projection cells. We found that the neostriatal interneurons can be divided at least into three classes by physiological, chemical and morphological criteria. The first was FS cells (fast-spiking cells) which fired very short-duration action potentials at constant spike frequency during depolarizing pulses, were immunoreactive for parvalbumin (calcium-binding protein), and had axons with very dense collateralization within or near their dendritic fields. Another class was identified as those which fired low-threshold spikes (LTS cells) from hyperpolarized potentials, were positive for somatostatin and nitric oxide synthase (NOS), and had the largest axonal fields. The other class of interneurons had longer-lasting afterhyperpolarizations (LA cells), were positive for choline acetyltransferase, and were mostly large aspiny cells. Glutamic acid decarboxylase (GAD67) or GABA immunoreactivity was detected at the somata or terminals of parvalbumin FS cells and somatostatin/NOS LTS cells, but not of cholinergic LA cells. Substance P, probably released from the collaterals of cells projecting to the substantia nigra, excited LA cells and LTS cells, but not FS cells. These results suggest that the striatum has at least one type of cholinergic and two types of GABAergic interneurons which are different in physiological, chemical and pharmacological characteristics.