Striatal cells containing aromatic L-amino acid decarboxylase: an immunohistochemical comparison with other classes of striatal neurons.

In a previous study, we described a population of striatal cells in the rat brain containing aromatic L-amino acid decarboxylase, the enzyme involved in the conversion of L-DOPA into dopamine. We have also presented evidence that these cells produce dopamine in the presence of exogenous L-DOPA. In this paper, we further characterize these striatal aromatic L-amino acid decarboxylase-containing cells in order to determine whether they form a subclass of one of the known categories of striatal neurons or if they represent a novel cell type. Using immunohistochemical methods, we compared the morphology and distribution of the aromatic L-amino acid decarboxylase-immunolabeled cells with those of other classes of striatal neurons. Our results show that both the morphology and distribution of aromatic L-amino acid decarboxylase-immunolabeled cells are very distinctive and do not resemble those of cells labeled for other striatal neuronal markers. Double-labeling procedures revealed that aromatic L-amino acid decarboxylase cells do not co-localize somatostatin or parvalbumin, and only a very small percentage of them co-localize calretinin. However, the population of aromatic L-amino acid decarboxylase cells label intensely for GABA.Overall, our results suggest that these aromatic L-amino acid decarboxylase-containing cells represent a class of striatal GABAergic neurons not described previously.

Altered long-term potentiation in the hippocampus of apolipoprotein E-deficient mice.

Recent studies suggest that apolipoprotein E (apoE) plays a neurotrophic role in the central nervous system and that an aberrant function of this molecule might result in neurodegeneration. Supporting this notion, apoE-deficient mice show neurodegenerative and cognitive alterations. To characterize physiological changes associated with synaptic damage and cognitive impairment in apoE-deficient mice, we investigated synaptic plasticity in the hippocampus of urethane anesthetized mice. Electrical stimulation was delivered to the perforant pathway and the resulting evoked field excitatory postsynaptic potential (EPSP) and population spike were recorded in the hilus. Long-term potentiation, as measured in the population spike, was reduced by 50% in apoE-deficient mice when compared to wild-type controls. In contrast, there were no significant differences in the evoked field EPSP between wild-type and apoE-deficient mice following high-frequency stimulation. These results support the notion that cognitive impairment and synaptic loss in the hippocampus of apoE-deficient mice might be associated with impaired long-term potentiation.

Lesions in the dentate hilum and CA2/CA3 regions of the rat hippocampus produce cognitive deficits that correlate with site-specific glial activation.

Thirty male rats pressed a lever three times (FR3) when a stimulus light (sD) was off to obtain sucrose pellets. They were then evenly divided into sham Controls versus groups lesioned bilaterally in the hippocampus by stercotaxic injection of ibotenic acid into the dentate hilum (HIL) or the CA2/CA3 region (CA2/3). On measures of recall of the FR3-sD task taken during the initial 30 min of a postlesion test session, the CA2/3 and especially the HIL groups showed significant (p < .05) impairments relative to the Controls. During an ensuing 30-min period, rats were reshaped to criterion, beginning at FR1, and no appreciable intergroup differences were noted on this schedule or at FR1-sD. At FR2-sD, the HIL but not the CA2/3 group showed some impairment relative to Controls. At FR3-sD, both the CA2/3 and HIL groups had impaired task performance. An immunocytochemical index of glial activation showed higher reactivity in CA2/3 or the dentate hilum among CA2/3 or HIL animals, respectively, that was associated with the degree to which they showed an FR3-sD performance deficit.

Glutamate-dependent long-term presynaptic changes in corticostriatal excitability.

We have previously shown that brief high frequency stimulation of the anteromedial prefrontal cortex induces a long-term decrease in excitability of the glutamatergic corticostriatal terminal field. In contrast, a long-term increase in presynaptic corticostriatal excitability may be induced by presenting two brief cortical tetanizing stimuli separated by 2-3 min such that the second tetanus coincides with a period of increased excitability elicited by the first. In the present study, we examined the glutamate receptor subtypes involved in these long-term changes in presynaptic excitability. A specific glutamate receptor antagonist was infused into the rat striatum 10-25 min prior to either a single or double cortical tetanic stimulation. To eliminate the participation of intrinsic striatal cells, a subset of animals received a striatal kainic acid lesion eight to 20 days before the recording experiment. Antagonists of the N-methyl-D-aspartate and metabotropic glutamate receptor subtypes were effective in blocking the decrease in excitability induced by single cortical tetanic stimulation whereas an antagonist of the alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid/kainate receptor did not prevent the induction of a long-term reduction in excitability. In contrast, each of these antagonists prevented the induction of a long-term increase in excitability. These long-term modifications in excitability of the presynaptic glutamate axon terminals appear to be induced by similar mechanisms to those postulated to operate in long-term potentiation and depression. These enduring changes in presynaptic excitability are likely to represent important mechanisms for the selective modification of information processing in the striatum.

Effects of nicotine on dopaminergic nigrostriatal axons requires stimulation of presynaptic glutamatergic receptors.

An electrophysiological method for evaluating changes in axonal excitability was used to examine presynaptic effects of the local striatal administration of nicotine on nigrostriatal dopaminergic terminal axons in the rat. To eliminate postsynaptic interactions, intrinsic striatal neurons were destroyed with a unilateral kainate lesion performed 10 to 15 days before the recording experiments. Excitability was assessed by determining the striatal stimulus current just sufficient to elicit an antidromic response from the striatal terminal field of a dopaminergic nigral neuron on 95% of the stimulus presentations. Local nicotine infusion (1-100 microM/0.3 microliter) produced a dose-dependent increase in excitability. Previous intrastriatal administration of the nicotine receptor antagonists mecamylamine or chlorisondamine blocked the nicotine effect and subsequent administration reversed the nicotine response. Increased dopamine autoreceptor stimulation, presumably resulting from nicotine-induced dopamine release, appeared to oppose the nicotine-induced increase in excitability. Accordingly, in animals in which dopamine synthesis was blocked with alpha-methyl-p-tyrosine (250 mg/kg, 12 and 2 h before recording), the nicotine-induced increase in terminal excitability was larger than in untreated rats. Simultaneous intrastriatal administration of the glutamate receptor antagonists, 6,7-dinitro-quinoxaline-2,3-dione and 2-amino-7-phosphonoheptanoate, prevented the nicotine-induced increase in excitability in animals with or without alpha-methyl-p-tyrosine pretreatment. We conclude that the nicotine-induced increase in nigrostriatal terminal excitability is an indirect effect resulting from a nicotine-evoked increase in glutamate release and a subsequent increase in the stimulation of presynaptic glutamate heteroreceptors on the dopamine-containing terminals.

Amphetamine-induced changes in nigrostriatal terminal excitability are modified following repeated amphetamine pretreatment.

To investigate neural mechanisms associated with behavioral sensitization to amphetamine, we studied the effect of an intrastriatal infusion of amphetamine on nigrostriatal axon terminal electrical excitability in rats following withdrawal from repeated systemic treatment. Rats were injected with amphetamine 2.5 mg/kg s.c. or saline daily for 4 days. Either 24 h or 14 days after the last injection, extracellular recordings were obtained from dopaminergic neurons of the substantia nigra, in a blind design in which the experimenter did not know the pretreatment regime. In order to assess the electrical excitability of the nigrostriatal axonal field, neurons were activated antidromically by stimulating their terminal fields in the striatum. As previously reported, striatal infusion of amphetamine (1 microM/0.3 microliter) in control animals resulted in a significant reduction in excitability as indicated by an increase in striatal stimulus current necessary to evoke antidromic activity. In contrast, intrastriatal amphetamine administration to amphetamine-pretreated animals did not decrease excitability. Spontaneous firing rates and patterns of cell discharge did not differ between saline- and amphetamine-treated animals. The chronic amphetamine-induced change in the effect of an acute intrastriatal amphetamine infusion on nigrostriatal terminal excitability may be due to enduring alterations in the amphetamine-induced release of dopamine and other striatal neurotransmitters or to changes in the sensitivity of presynaptic hetero- and/or autoreceptors on the dopaminergic axons.

Aromatic L-amino acid decarboxylase immunoreactive cells in the rat striatum: a possible site for the conversion of exogenous L-DOPA to dopamine.

The efficacy of L-dihydroxyphenylalanine (L-DOPA) in ameliorating the symptoms of Parkinson's disease (PD) is attributed to its conversion to dopamine (DA) by the enzyme aromatic L-amino-acid decarboxylase (AADC) in the striatum. Although the site of this conversion in the DA-denervated striatum has yet to be identified, it has been proposed that L-DOPA could be converted to DA at non-dopaminergic sites containing AADC. In the present study, we used immunocytochemical techniques to examine the localization of AADC and DA in the striatum of rats with a unilateral 6-hydroxydopamine (6-OHDA) lesion of the nigrostriatal dopaminergic projection. In the DA-denervated striatum, we observed AADC-immunoreactive (-IR) cells with morphological characteristics similar to a class of small aspiny interneuron. Although usually obscured by a dense plexus of AADC-IR fibers, these cells could also occasionally be detected in the intact striatum. Acute administration of L-DOPA to DA-denervated animals elicited contralateral rotational behavior as well as a pronounced c-fos protein immunoreactivity in the striatum ipsilateral to the lesion. Following acute administration of L-DOPA, but not after acute saline, DA-IR cells were detected in the denervated striatum. These DA-IR cells are similar in morphology and were found in the same location as the AADC-IR cells. These results strongly suggest the existence of a class of AADC-containing striatal cells that can form DA from exogenous L-DOPA in the rat. In the DA deafferented striatum, DA produced by these cells from exogenous L-DOPA could be released to exert physiological effects on DA receptive tissue. It is possible that similar cells could contribute to the efficacy of L-DOPA in the treatment of Parkinson's disease.

Electron microscopic evidence for neurotoxicity in the basal ganglia.

The dopaminergic projection from the substantia nigra to the neostriatum is vulnerable to several neurotoxins including 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), amphetamine, and 5-hydroxydopamine. We have treated rats or mice with these agents and examined various regions of their brains with a combination of Fink-Heimer, immunohistochemical, serial-section electron microscopic, and three-dimensional reconstruction methods. In addition to degenerating or swollen axons, we found darkened glial processes and some damage to postsynaptic cells and dendrites. The particular effects observed critically depend on experimental variables such as dose, time, species and strain and raise questions about the correlation of light and electron microscopic results. These studies provide the basis for a discussion of the advantages and disadvantages of an ultrastructural examination of the effects of neurotoxins.

Cerebellar-responsive neurons in the thalamic ventroanterior-ventrolateral complex of rats: in vivo electrophysiology.

In vivo intracellular recordings were obtained from identified thalamocortical neurons in the ventroanterior-ventrolateral complex in urethane-anesthetized rats. This thalamic nucleus has few interneurons. Neurons that responded to cerebellar stimulation were injected intracellularly with horseradish peroxidase or biocytin and examined with light and electron microscopy (see companion paper). Intrinsic membrane properties and voltage-dependent rhythmic activity of cerebellar-responsive ventroanterior-ventrolateral neurons were similar to those described previously for thalamic neurons. Thus, in addition to conventional "fast" Na(+)-dependent spikes, rat ventroanterior-ventrolateral neurons had "slow" Ca(2+)-mediated low-threshold spikes and membrane conductances that supported rhythmic oscillations. Two modes of spontaneous activity were observed: (i) a tonic firing pattern that consisted of irregularly occurring fast spikes that predominated when the membrane potential was more positive than about -60 mV, and (ii) a rhythmic firing pattern, observed when the membrane potential was more negative than about -65 mV, composed of periodic (4-8 Hz) membrane hyperpolarizations and ramp depolarizations that often produced a low-threshold spike and a burst of fast spikes. In some neurons, spontaneous fast prepotentials were also observed, often with a relatively constant rate (up to 70 Hz). Cerebellar stimulation elicited excitatory postsynaptic potentials that in some cases appeared to be all-or-none and were similar in form to fast prepotentials. Stimulation of ipsilateral motor cortex elicited a short-latency antidromic response followed by a monosynaptic excitatory postsynaptic potential, which had a slower rise time than excitatory postsynaptic potentials evoked from cerebellum, suggesting that cortical inputs were electrotonically distal to cerebellar inputs. In the presence of moderate membrane hyperpolarization, the cortically evoked excitatory postsynaptic potential was followed by a long-lasting hyperpolarization (100-400 ms duration), a rebound depolarization and one or two cycles resembling spontaneous rhythmic activity. Membrane conductance was increased during the initial component of the long hyperpolarization, much of which was probably due to an inhibitory postsynaptic potential. In contrast, membrane conductance was unchanged or slightly decreased during the latter three-quarters of the long hyperpolarization. The amplitude of this component of the long hyperpolarization usually decreased when the membrane was hyperpolarized with intracellular current injection. Thus, both disfacilitation and an inhibitory postsynaptic potential may have contributed to the latter portion of the cortically-evoked long hyperpolarization. The cortically-evoked inhibitory postsynaptic potentials likely originated predominantly from feedforward activation of GABAergic neurons in the thalamic reticular nuclei.(ABSTRACT TRUNCATED AT 400 WORDS)

Cerebellar-responsive neurons in the thalamic ventroanterior-ventrolateral complex of rats: light and electron microscopy.

The morphology and synaptic organization of neurons in the ventroanterior-ventrolateral nucleus of rats was examined using in vivo intracellular staining techniques. Neurons were characterized electrophysiologically based on intrinsic membrane properties and synaptic responses to stimulation of motor cortex and cerebellar nuclei, as described in the companion paper. Cerebellar-responsive neurons were stained intracellularly with either horseradish peroxidase or biocytin. All stained ventroanterior-ventrolateral nucleus neurons were identified as thalamocortical neurons on anatomical (and often electrophysiological) grounds, consistent with previous findings that rat ventroanterior-ventrolateral nucleus is interneuron-sparse. Ventroanterior-ventrolateral nucleus neurons had three to eight thick primary dendrites. Proximal dendrites often exhibited a tufted branching pattern, from which many thinner, higher order dendrites arose. Dendrites branched to form a funnel-like infiltration of the neuropil that resulted in a spherical, roughly homogeneous dendritic field. The axon originated from the cell body or a proximal dendrite and coursed laterally and dorsally to innervate motor cortex. One to five axon collaterals were emitted in the rostral dorsolateral sector of the thalamic reticular nucleus; collaterals were not observed in the ventroanterior-ventrolateral nucleus or other nuclei in dorsal thalamus. The synaptic organization of the ventroanterior-ventrolateral nucleus was examined with electron microscopy, including two intracellularly labeled ventroanterior-ventrolateral nucleus neurons that were shown electrophysiologically to receive monosynaptic inputs from the cerebellum. The neuropil of rat ventroanterior-ventrolateral nucleus lacked the complexity and diversity found in corresponding thalamic nuclei of felines and primates, due to the paucity of interneurons. Vesicle-containing dendrites, dendrodendritic synapses and glomeruli were not observed. Three broad classes of presynaptic terminals were identified. (1) Small round boutons: small boutons containing densely-packed, small round vesicles that formed asymmetric synapses predominantly with the distal dendrites of thalamocortical neurons. These were the most prevalent type of bouton in the ventroanterior-ventrolateral nucleus (78% of presynaptic elements) and likely arose from the cerebral cortex. (2) Large round boutons: large terminals with loosely packed small round vesicles that made multiple asymmetric synapses with proximal and intermediate dendrites. Large round boutons comprised 8% of the neuropil, and likely arose from the cerebellar nuclei. (3) Medium size boutons with pleomorphic vesicles: medium-sized profiles containing pleomorphic vesicles that formed symmetric synapses with proximal, intermediate and distal dendrites and, less frequently, with cell bodies.(ABSTRACT TRUNCATED AT 400 WORDS)

The distribution of cholinergic perikarya with respect to enkephalin-rich patches in the caudate nucleus of the adult cat.

The distribution of cholinergic interneurons with respect to enkephalin-rich patches in the caudate nucleus of the cat was examined using both computer-assisted 3-D reconstruction and immunocytochemical techniques. Examination of the 3-D distribution of perikarya staining for choline acetyltransferase (ChAT) revealed that these cells were not evenly distributed within the caudate nucleus but exhibited areas of increased and decreased density. Comparison of the 3-D distribution of cholinergic perikarya to that of the enkephalin-rich patches indicated that areas of increased ChAT+ cell density often corresponded to the positions of enkephalin-rich patches within the dorsal-lateral caudate nucleus. At more ventral regions, there was no clear correspondence between areas of increased ChAT+ cell density and enkephalin-rich patches. In agreement with these observations, a quantitative analysis of sections double-labeled for ChAT and enkephalin revealed that the density of cholinergic neurons within enkephalin-rich patches was twice that in the surrounding tissue in the dorsal region of the caudate nucleus. In contrast at more ventral levels, the difference in the density of ChAT+ cells in enkephalin-rich patches did not significantly differ from that in the surrounding striatal tissue. Both the results of the 3-D and the double-labeling analysis suggest that cholinergic neurons are not evenly distributed within the caudate nucleus of the cat but form loose clusters which are associated dorsally with the enkephalin-rich patches. These results also provide further evidence of heterogeneity within the striosomal compartment in the cat.

Compartmental organization of the peptide network in the human caudate nucleus.

The mammalian striatum may be divided into a striosomal compartment and a surrounding matrix region. We have examined the distribution of leucine enkephalin (LENK) and substance P (SP) immunoreactivity in relation to striosomes defined by calbindin-D (CABD) staining in alternate 70 microns serial sections from the human caudate nucleus. The distribution of LENK immunoreactivity showed a transition from dorsal to ventral striatum: dorsally, LENK-rich patches were present in a lightly stained matrix; mid-ventrally, annular patches of LENK staining with a lighter core were seen. These patches corresponded to striosomal regions defined by CABD-poor zones. In contrast, in the ventral caudate and nucleus accumbens, LENK-poor zones matched CABD-defined striosomes. CABD staining in the matrix was intense in the dorsal caudate, diminishing ventrally. SP-rich zones in dorsal caudate and SP-poor areas in the mid-ventral region overlapped striosomes. In the ventromedial sector, the SP staining pattern was complex and did not consistently correlate with striosomes. Computer-assisted three-dimensional reconstruction of the striosomal system in the human, based on regions of either high LENK or low CABD immunoreactivity, revealed the existence of considerable long-range order. Patches appeared aligned over several millimeters to form long, horizontal structures in the caudate nucleus, with occasional orthogonal interconnecting crossbridges. Our results are in accord with previous work in the human and in other species. These three-dimensional networks are strikingly similar across individuals and may relate to the segregation of and interactions between striatal circuits.

5-hydroxydopamine-labeled dopaminergic axons: three-dimensional reconstructions of axons, synapses and postsynaptic targets in rat neostriatum.

Previous studies employing 5-hydroxydopamine to identify nigrostriatal dopaminergic axons and their synapses found that labeled axons made few synapses or that asymmetric contacts predominated. In contrast, recent studies using tyrosine hydroxylase or dopamine antibody techniques indicate that presumed dopaminergic axons form small symmetric contacts. We re-examined 5-hydroxydopamine-labeled material from the rat neostriatum using serial three-dimensional reconstruction techniques to characterize the morphology of labeled axons, synapses and postsynaptic targets. This ultrastructural analysis revealed a class of heavily labeled axons that are small (0.06-1.5 microns in diameter) and lack large varicosities. These axons form small (0.011-0.09 microns 2), en passant, symmetric synapses, mainly onto dendritic spines and spiny dendritic shafts and, in some cases, onto aspiny dendritic segments near branch points. The sites of these synapses along the axon appeared unrelated to the locations of axonal enlargements, suggesting that counting varicosities may not be an accurate indication of the extent of dopaminergic innervation in the neostriatum. The characteristics of these 5-hydroxydopamine-labeled elements correspond in all respects to axons and synapses identified as dopaminergic by immunohistochemistry in previous studies. In tissue in which all labeled and unlabeled synapses were classified, approximately 9% of all synapses were identified as dopaminergic by this type of label. Three-dimensional reconstructions provided additional insight concerning the interaction of dopaminergic afferents with postsynaptic striatal targets and their relation to other afferents to these neurons. They reveal that a short, unbranched dopaminergic axonal segment can make multiple synapses onto dendritic spines, shafts and branch points of one or more dendrites. In addition, one dendrite can receive contacts from several labeled axons. Dopamine synapses onto spines are always associated with unlabeled, asymmetric synapses onto the same spine. Synapses of various morphologies with a distinctly different, lighter form of labeling were much rarer, and may represent other aminergic afferents to the neostriatum. The presence of this second form of label in earlier 5-hydroxydopamine studies may have contributed to the long-standing controversy over the appearance of dopaminergic synapses examined by different techniques. Our results help to resolve this controversy and confirm that the nigrostriatal projection makes small symmetric synapses with a variety of striatal targets.

Cholinergic neurons are distributed preferentially in areas rich in substance P-like immunoreactivity in the caudate nucleus of the adult cat.

The distribution of cells stained immunocytochemically for the cholinergic marker choline acetyltransferase was compared to the pattern of substance P immunoreactivity in the caudate nucleus of adult cats using a double-label immunocytochemical protocol and three-dimensional reconstructions of adjacent sections single-labeled for either substance P or choline acetyltransferase. Substance P immunoreactivity was distributed in a highly complex mosaic within the caudate nucleus of the cat. In the dorsal caudate nucleus, substance P-rich zones consisting of either clusters of substance P-positive cell bodies or fibers were seen against a lighter staining background. The density of cholinergic neurons was found to be significantly greater within these substance P-rich patches in comparison to surrounding regions. The pattern of substance P immunoreactivity within the ventral caudate nucleus differed from that in more dorsal regions. Clear substance P-rich patches were not seen in this region, but a large substance P-rich area consisting of a dense plexus of substance P-containing fibers was visible. Embedded within this substance P-rich area were fairly discrete patches of light substance P staining. As in the dorsal caudate nucleus, increased numbers of cholinergic neurons and processes were associated with substance P-rich regions in the ventral caudate nucleus. Choline acetyltransferase-positive perikarya also appeared to be concentrated in substance P-rich areas in the nucleus accumbens and olfactory tubercle. The results of this study suggest that a close relationship exists between the distribution of substance P fibers and cholinergic perikarya in the striatum of the cat.

Ultrastructural examination of enkephalin and substance P input to cholinergic neurons within the rat neostriatum.

Enkephalin and substance P-containing inputs to cholinergic perikarya were examined in the rat neostriatum using an ultrastructural immunocytochemical double-labeling protocol. Sections of rat neostriatum were double-labeled for either choline acetyltransferase (ChAT) and substance P or ChAT and enkephalin using silver intensified colloidal gold and peroxidase as labels. Regions containing both ChAT-positive neurons and peroxidase reaction product were identified in the light microscope prior to sectioning for electron microscopy. Substance P-containing terminals which contained round synaptic vesicles and made symmetrical synaptic contacts were commonly observed in the neostriatum. Substance P synapses onto ChAT-positive perikarya and dendrites were frequently observed: up to 5 synaptic contacts were observed onto a ChAT-positive dendrite. Enkephalin labeling was also seen in a population of axon terminals containing round synaptic vesicles and exhibiting symmetrical synaptic specializations. In contrast to substance P-containing terminals, relatively few synaptic contacts were observed onto ChAT-positive labeled perikarya and dendrites although enkephalin-labeled terminals were seen in frequent contact with perikarya and dendrites of unlabeled spiny neurons. Since enkephalin and substance P are contained within different populations of striatal spiny neurons, the results of the present study suggest that these two types of neurons differ in their intrinsic striatal connections.

Effects of frequency and pattern of medial forebrain bundle stimulation on caudate dialysate dopamine and serotonin.

In vivo microdialysis was employed to detect changes in extracellular dopamine and serotonin in the rat caudate in response to electrical stimulation of the medial forebrain bundle. Extracellular dopamine concentrations increased linearly as a function of the frequency (4-33 Hz) of evenly spaced stimuli in both the presence and absence of cocaine added to the dialysate. Because dopamine neurons are known to fire in single-spike and burst patterns, stimulation pulses were also delivered in a bursting pattern. The response of extracellular dopamine was augmented in both the presence and absence of cocaine when the same number of stimuli were delivered in bursts as compared to an evenly spaced pattern. Serotonin, which was only assessed in the presence of cocaine, similarly increased linearly with frequency, but, in contrast to the dopamine response, levels of serotonin were not augmented by stimuli presented in bursts. These results suggest that microdialysis can be used to detect physiological changes in synaptic transmitter concentrations.

Presynaptic long-term changes in excitability of the corticostriatal pathway.

We employed measurements of striatal terminal excitability to monitor the presynaptic effects of tetanic stimulation of corticostriatal fibers. Cortical tetanic stimulation (CTS) initiated a long-lasting decrease in terminal excitability. With higher current CTS, a transient increase in excitability preceded the decrease. However, a long-term increase was induced (1) by a second tetanus applied during the brief elevation in excitability initiated by a previous CTS or (2) when dopamine and GABA transmission were disrupted. A long-term increase also occurred following tetanic stimulation of the striatal terminal field (STS). The direction of the long-lasting change in excitability may depend on the level of polarization of the membrane. These presynaptic mechanisms could be important for the long-term selective modification of striatal synaptic transmission.

Terminal excitability of the corticostriatal pathway. I. Regulation by dopamine receptor stimulation.

Glutamatergic cortical and dopaminergic nigral afferents converge onto neurons of the neostriatum forming synapses in close proximity. Studies, mainly using pharmacological methods, suggest presynaptic interactions between these afferents. The influence of dopaminergic transmission on the cortical terminal fields in the striatum was assessed electrophysiologically using the terminal excitability method. Antidromic action potentials recorded from neurons in the prefrontal cortex were elicited by bipolar electrical stimulation (250 microns wire, 0.5 mm tip separation) of the cortical terminal field in the contralateral dorsomedial neostriatum. Threshold excitability was defined as the minimum current sufficient to elicit 95-100% antidromic response on non-collision trials. Under control conditions, the mean threshold current was 1.7 +/- 0.2 mA. Drugs were applied in a volume of 312 nl delivered over 5 min to the striatal stimulation site. Following local striatal administration of amphetamine (10 microM) or electrical stimulation of the nigrostriatal pathway (1-2 pulses, 1.5 mA/0.5 ms/1 Hz) an increase in striatal stimulating current was required in order to reinstate threshold levels of antidromic response. This decrease in the excitability of corticostriatal afferents could be reversed by local infusion of haloperidol (1 microM) or L-sulpiride (10 nM) and did not occur following depletion of dopamine stores with alpha-methylparatyrosine and reserpine. The possible participation of postsynaptic dopamine receptor stimulation was ruled out as these effects were still seen in animals with kainic acid induced lesions of the striatum. In addition, terminal excitability was not modified by the muscarinic agonist carbachol (10 microM). Striatal administration of apomorphine (10 microM) decreased terminal excitability similar to amphetamine. The specific D-2 agonist, quinpirole (10-20 microM) did not affect excitability. These results indicate that manipulations which have been shown to increase the release of endogenous dopamine decrease the excitability of prefrontal corticostriatal afferents by stimulation of presynaptic dopamine receptors which are insensitive to low doses of quinpirole but sensitive to L-sulpiride and apomorphine. The mechanisms underlying dopamine-induced changes in terminal excitability are likely to be similar to those which have been shown to alter conductance at postsynaptic sites.

Terminal excitability of the corticostriatal pathway. II. Regulation by glutamate receptor stimulation.

The influence of impulse activity and glutamate receptor stimulation on the electrical excitability of the corticostriatal terminal field was explored. Antidromic responses were recorded from prefrontal cortical neurons the electrical stimulation of their terminal field in the contralateral striatum. Terminal excitability was assessed by determining the percentage of subthreshold current stimulus presentations eliciting an antidromic response. Terminal excitability was found to be positively correlated with variations in spontaneous firing rate: increases and decreases in firing rate were accompanied by corresponding changes in the percentage of antidromic responses elicited by a subthreshold stimulus. Drugs were applied to the striatal stimulation site in a volume of 312 nl delivered over 5 min. Striatal administration of either the competitive NMDA antagonist D-alpha-aminoadipate (DAA) or D-2-amino-7-phosphonoheptanoate (AP-7) or the competitive non-NMDA antagonist 6-cyano-7-nitroquinoxaline-2,3 dione (CNQX) blocked the correlation between excitability and firing rate. Further examination revealed that the terminal field was rendered more excitable for a period of 20-80 ms following the arrival of an action potential. This post-impulse facilitation of terminal excitability was attenuated after local application of AP-7 (10 microM) or CNQX (20 microM). At half these doses, AP-7 or CNQX produced a non-significant effect, however when administered simultaneously a significant attenuation was observed. The participation of interneurons in these excitability effects was ruled out since they were still seen following kainic acid lesions. We propose that this impulse-dependent enhancement in terminal excitability results from the release of glutamate induced by the action potential in the terminal field and the subsequent stimulation of glutamate autoreceptors on the terminals.

Electrophysiological characteristics of cells within mesencephalon suspension grafts.

Both spontaneous and evoked extracellular electrophysiological activity of neurons within fetal mesencephalon suspension grafts to the dopamine-depleted striatum of rats were examined. In some cases, extracellular recording was combined with intracellular labeling to identify recorded neurons. Grafted rats displaying a complete cessation of ipsilateral rotations following amphetamine administration were examined at post-implantation time intervals of two, four, five, eight and nine months. Four separate classes of neurons were distinguished within the transplanted striatum based on electrophysiological properties. The first of these groups, the type I cells, appeared to be non-grafted striatal neurons. When spontaneously active, these striatal-like cells fired bursts of action potentials separated by periods of decreased activity. Evoked responses in these cells were characteristic of striatal cells. Type I cells which were intracellularly labeled were found outside the grafts and displayed the characteristic morphology of the medium spiny neuron of the neostriatum. The other three cell classes displayed electrophysiological properties similar to neurons recorded in situ within the reticular formation, substantia nigra pars compacta and substantia nigra pars reticulata. Neurons from these three groups which were labeled with an intracellular marker were found to lie within the suspension grafts. The spontaneous activity of the pars compacta dopaminergic-like neurons was predominantly irregular, with some cells also firing in a regular or pacemaker-like pattern. Infrequently, irregular firing dopaminergic-like neurons displayed episodes of doublet bursting. Many of the grafted neurons responded to electrical stimulation of prefrontal cortex and striatum, indicating that the graft was receiving functional inputs from host neurons. Comparison of the firing rate and pattern of grafted neurons to in situ mesencephalic neurons as a function of time following grafting suggested that the grafted neurons and/or the neuronal circuitry is slowly developing within the host environment. A prolonged time-course for the maturation of the graft may be reflected in the time required to achieve improvements in some behavioral deficits following transplantation. However, the relatively rapid recovery of drug-induced rotational asymmetry following grafting suggests that this form of recovery may not require mature functioning of the grafted neurons.