Nigral dopamine neurons: intracellular recording and identification with L-dopa injection and histofluorescence.

Intracellular recordings in vivo were obtained from dopamine-containing neurons of the rat substantia nigra. These neurons were identified electrophysiologically by antidromic activation and histochemically by L-dopa injection and subsequent fluorescence histochemistry. Extracellular spikes and antidromic conduction velocity of the neurons were identical to those previously described for putative dopaminergic neurons. Spontaneous intracellular fast potentials, slow depolarizations during burst firing, and spike prepotentials were observed.

Proglumide: selective antagonism of excitatory effects of cholecystokinin in central nervous system.

Extracellular single-unit recording techniques were used to test the ability of proglumide to block cholecystokinin-induced excitation of rat midbrain dopaminergic neurons and dopamine-sensitive prefrontal cortex cells. Intravenous and iontophoretic proglumide administration consistently blocked cholecystokinin-induced excitations while having no effect on glutamic acid-induced increases in activity. This selective blockade of central cholecystokinin effects by proglumide suggests that this drug may be valuable for studying the possible role of cholecystokinin as a neurotransmitter or neuromodulator in the central nervous system.

d-Amphetamine-induced inhibition of central dopaminergic neurons: mediation by a striato-nigral feedback pathway.

Lesions of the striato-nigral pathways (that is, crus cerebri or vicinity of the tail of the caudate nucleus) markedly attenuate depressant effects of intravenous d-amphetamine on central dopaminergic cell activity. These results, coupled with previous data showing that microiontophoretic application of d-amphetamine directly onto dopaminergic cells does not produce significant slowing, provide direct support for the hypothesis that the depressant effect of d-amphetamine on these cells is mediated through a striato-nigral neuronal feedback loop.

Dopamine auto- and postsynaptic receptors: electrophysiological evidence for differential sensitivity to dopamine agonists.

The responses of dopamine cells in the substantia nigra to iontophoretically administered dopamine and intravenous apomorphine were compared to the responses of spontaneously active neurons in the caudate nucleus. Dopaminergic cells were six to ten times more sensitive to dopamine and intravenous apomorphine than 86 percent of the caudate cells tested. This differential sensitivity of dopamine auto- and postsynaptic receptors may explain the apparently paradoxical behavioral effects induced by small compared to large doses of some dopamine agonists and may provide a means of developing new types of drugs to antagonize dopaminergic influence in the central nervous system.

Dopamine-cell depolarization block as a model for the therapeutic actions of antipsychotic drugs.

Antipsychotic drugs used in the treatment of schizophrenia have in common the property of being dopamine-receptor antagonists. However, the rapid timecourse of receptor blockade produced upon drug administration does not correlate with the emergence of clinical actions, which typically require weeks of treatment to become manifest. Studies in rats have shown that repeated antipsychotic drug treatment results in a delayed inactivation of dopamine-neuron firing in the midbrain due to depolarization block. Furthermore, the therapeutic efficacy of antipsychotic drugs in humans correlates with their ability to induce depolarization block of mesolimbic dopamine neurons, whereas their potential to produce extrapyramidal side effects correlates with their propensity for inducing depolarization block in the nigrostriatal dopamine system. Therefore, dopamine-cell depolarization block is an effective model for evaluating antipsychotic drug efficacy, and provides a potential mechanism to account for their therapeutic impact on a dysregulated dopamine system.

Dual effects of D-amphetamine on dopamine neurons mediated by dopamine and nondopamine receptors.

By increasing dopamine (DA) release and activating feedback mechanisms, amphetamine and related psychostimulants are known to inhibit DA cell firing. Here, we report that D-amphetamine also has an excitatory effect on DA cells, which under control conditions, is masked by the inhibitory effect of D-amphetamine and is revealed when D2-like receptors are blocked. Thus, using in vivo single-unit recording in rats, we found that the selective D2 antagonist raclopride not only blocked the inhibition induced by D-amphetamine but also enabled D-amphetamine to excite DA cells. The excitation, expressed as an increase in both firing rate and bursting, persisted when both D1- and D2-like receptors were blocked by SCH23390 and eticlopride, suggesting that it is not mediated by DA receptors. The norepinephrine uptake blocker nisoxetine mimicked the effect of D-amphetamine, especially the increase in bursting, whereas the 5-HT uptake blocker fluoxetine produced no significant effect. Adrenergic alpha1 antagonists prazosin and WB4101 and the nonselective alpha antagonist phenoxybenzamine completely blocked increase in bursting induced by D-amphetamine and partially blocked the increase in firing rate. The alpha2 antagonist idazoxan and the beta antagonist propranolole, however, failed to prevent D-amphetamine from producing the excitation. Thus, revising the traditional concept, this study suggests that D-amphetamine has two effects on DA cells, a DA-mediated inhibition and a non-DA-mediated excitation. The latter is mediated in part through adrenergic alpha1 receptors.

Psychiatric status after human fetal mesencephalic tissue transplantation in Parkinson's disease.

This report describes the prospective and systematic psychiatric assessment of nine patients who received transplantation of human fetal mesencephalic tissue into the caudate nucleus for treatment of Parkinson's disease. Unlike adrenal medullary transplantation, which often causes psychosis or delirium, this procedure appeared to have few perioperative sequelae. On longer-term follow-up, there was some statistical evidence of deterioration in psychiatric status, as manifested primarily in depressive and nonspecific emotional and behavioral symptoms. This group effect was partly attributable to the occurrence of discrete episodes of illness (major depression and panic disorder with agoraphobia) in some patients, but it was unclear whether such episodes occurred more often than would ordinarily be expected in Parkinson's disease. Differences in the neurobiological effects of fetal mesencephalic and adrenal medullary grafts may account for differences in the psychiatric sequelae of patients receiving these procedures.

Topographical organization of the efferent projections of the medial prefrontal cortex in the rat: an anterograde tract-tracing study with Phaseolus vulgaris leucoagglutinin.

The purpose of the present investigation was to examine the topographical organization of efferent projections from the cytoarchitectonic divisions of the mPFC (the medial precentral, dorsal anterior cingulate and prelimbic cortices). We also sought to determine whether the efferents from different regions within the prelimbic division were organized topographically. Anterograde transport of Phaseolus vulgaris leucoagglutinin was used to examine the efferent projections from restricted injection sites within the mPFC. Major targets of the prelimbic area were found to include prefrontal, cingulate, and perirhinal cortical structures, the dorsomedial and ventral striatum, basal forebrain nuclei, basolateral amygdala, lateral hypothalamus, mediodorsal, midline and intralaminar thalamic nuclei, periaqueductal gray region, ventral midbrain tegmentum, laterodorsal tegmental nucleus, and raphe nuclei. Previously unreported projections of the prelimbic region were also observed, including efferents to the anterior olfactory nucleus, the piriform cortex, and the pedunculopontine tegmental-cuneiform region. A topographical organization governed the efferent projections from the prelimbic area, such that the position of terminal fields within target structures was determined by the rostrocaudal, dorsoventral, and mediolateral placement of the injection sites. Efferent projections from the medial precentral and dorsal anterior cingulate divisions (dorsomedial PFC) were organized in a similar topographical fashion and produced a pattern of anterograde labeling different from that seen with prelimbic injection sites. Target structures innervated primarily by the dorsomedial PFC included certain neocortical fields (the motor, somatosensory, and visual cortices), the dorsolateral striatum, superior colliculus, deep mesencephalic nucleus, and the pontine and medullary reticular formation. Previously unreported projections to the paraoculomotor central gray area and the mesencephalic trigeminal nucleus were observed following dorsomedial PFC injections. These results indicate that the efferent projections of the mPFC are topographically organized within and across the cytoarchitectonic divisions of the medial wall cortex. The significance of topographically organized and restricted projections of the rat mPFC is discussed in light of behavioral and physiological studies indicating functional heterogeneity of this region.

Effects of lesions in the medial prefrontal cortex on the activity of midbrain dopamine neurons.

To determine whether lesions in the prefrontal cortex (PFC) alter the activity of midbrain dopamine (DA) neurons, single unit recordings were made from DA neurons in control and lesioned rats. PFC lesions, obtained by local injection of ibotenic acid into the medial PFC, had no effect on either firing rate or bursting activity of DA neurons in the ventral tegmental area (VTA). However, the number of spontaneously active DA neurons in the VTA was significantly decreased. In the substantia nigra (SN), the same lesions increased the firing rate and had no effect on either the bursting activity of the number of active DA cells. These results suggest that PFC lesions alter the activity of DA neurons. However, VTA and SN DA neurons may respond differently to PFC lesions.

Central dopamine-peptide interactions: electrophysiological studies.

A variety of peptides have been shown to have physiological activity, either pre- or post-synaptically in DA systems. Significantly, this has not been true of all the peptides tested and physiological activity has for the most part, correlated with anatomical evidence for the presence of that peptide in the same area as DA and with the presence of specific receptors for the active peptide. In the main, biochemical and behavioral studies of the effects of peptides on the function of DA systems also correlate well with the effects of the various peptides on the activity of DA neurons. However, despite this growing body of knowledge we still know very little about the physiological relevance of most peptides to animal behavior or the interaction between these peptides and DA systems. This is due, in part, to the fact that the functioning of peptide systems is still relatively difficult to study in terms of synthesis, release, metabolism and turnover. The fact that there are very few selective non-peptide antagonists for many of these substances has also hampered the study of their function in the brain. Nevertheless, their proven interaction with midbrain DA systems, regardless of whatever else they may do in the brain, makes them of great potential clinical interest. Given the number of neurological and mental disorders in which malfunction of the DA system is presumed to be involved, any endogenous substance that can modulate activity of the DA cells and thus provide a new mechanism for controlling the functioning of central DA systems may have potential therapeutic value.(ABSTRACT TRUNCATED AT 250 WORDS)

Neurotensin attenuates dopamine D2 agonist quinpirole-induced inhibition of midbrain dopamine neurons.

The effects of intracerebroventricular (i.c.v.) administration of NT on the activity of midbrain DA neurons were studied in rats using single unit recording techniques. NT (i.c.v., 50 micrograms) was found to have no significant effect on the spontaneous activity of DA cells. On the other hand, NT treatment significantly attenuated the inhibitory effect of quinpirole (i.v.), a specific D2 agonist, on a subpopulation of DA cells. This result is consistent with previous behavioral and biochemical studies suggesting that NT may produce some of its effects through modulation of central DA systems.

Electrophysiological studies on EMD 23 448, an indolyl-3-butylamine, in the rat: a putative selective dopamine autoreceptor agonist.

Single unit electrophysiological and microiontophoretic techniques were used to study the effects of a putative selective dopamine autoreceptor agonist, EMD 23 448, an indolyl-3-butylamine on the activity of both dopamine neurons in the substantia nigra and dopamine-sensitive neurons in the caudate nucleus. Intravenous administration of EMD 23 448 inhibited the firing of dopamine neurons in the nigra in a dose-dependent manner. This effect could be readily reversed by the subsequent intravenous administration of the dopamine receptor antagonist, haloperidol. Microiontophoretic application of EMD 23 448 onto these neurons also inhibited their activity and this effect was blocked by iontophoresis of the dopamine receptor antagonist, trifluoperazine. Intravenous administration of the former compound increased the activity of dopamine-sensitive cells in the caudate in the same doses which inhibited dopamine neurons in the nigra. However, this effect appeared to be indirect since the iontophoretic application of EMD 23 448 directly onto caudate neurons had no effect on their firing rate. In addition, the activity of norepinephrine-containing neurons in the locus coruleus and serotonin-containing cells in the dorsal raphé nucleus was not changed by direct microiontophoretic application of this compound. In contrast, EMD 23 448 selectively blocked the acetylcholine-induced activation of pyramidal neurons in the hippocampus. These findings provide further evidence that with respect to central catecholamine neurons EMD 23 448 may be a selective agonist for dopamine autoreceptors.

Intracellular and extracellular electrophysiology of nigral dopaminergic neurons--1. Identification and characterization.

Intracellular recordings were obtained from directly identified rat nigral dopamine cells in vivo. This identification was based on an increase in glyoxylic acid-induced catecholamine fluorescence in the impaled dopamine neurons. One of three compounds was injected intracellularly into each cell to produce the heightened fluorescence: (1) L-DOPA, to increase the intracellular dopamine content by precursor loading; (2) tetrahydrobiopterin, a cofactor for tyrosine hydroxylase, to increase intracellular dopamine concentration through activation of the rate-limiting enzyme for dopamine synthesis and (3) colchicine, to arrest intraneuronal transport and thus allow the build-up of dopamine synthesizing enzymes and dopamine in the soma. In addition, dopamine cells were antidromically activated from the caudate nucleus and collision with a directly elicited action potential was demonstrated. Identified dopamine neurons were shown to possess an input resistance of 31.2 +/- 7.4 M omega (means +/- SD) and a time constant of 12.1 +/- 3.2 ms. The action potentials were of long duration (2.75 +/- 0.5 ms) with a marked break between the initial segment and the somatodendritic spike components. The initial segment was the only component commonly elicited during antidromic activation. Spontaneously occurring action potentials were usually preceded by a slow, pacemaker-like depolarization. Burst firing by summation of depolarizing afterpotentials was observed to occur spontaneously, but could not be triggered by short depolarizing current pulses. Intravenously administered apomorphine demonstrated the same inhibitory effect on cell firing that was previously reported to occur when recording extracellularly from identified dopaminergic neurons. The determination of the electrophysiological characteristics of a population of cells directly identified as containing a specific neurotransmitter (in this case, dopamine) may allow one to construct better models of a system's functioning. Thus, the high input resistance and long time constant of dopamine-containing cells, combined with their burst/pause firing mode, may be important functionally with respect to a possible modulatory effect of dopamine in postsynaptic target areas.

Intracellular and extracellular electrophysiology of nigral dopaminergic neurons--2. Action potential generating mechanisms and morphological correlates.

Intracellular recordings from identified nigral dopamine neurons in the rat revealed that their potentials are composed of four components: (1) a slow depolarization, (2) an initial segment spike, (3) a somatodendritic spike, and (4) an afterhyperpolarization. By combining intracellular and extracellular recording techniques with anatomical studies using intracellular injections of Lucifer yellow, an attempt was made to localize each of these potentials to various neuronal compartments. Lucifer yellow injections demonstrated that the dopamine neurons recorded have a pyramidal or polygonal shaped soma, 12-30 microns in diameter, with 3-6 thick major dendrites which extend 10-50 microns from the soma before bifurcating. The axon appears to rise from a major dendrite 15-30 microns from the soma. Based on this anatomical configuration, results from the electrophysiological studies suggest that: (1) the slow depolarization is a pacemaker-like conductance most likely localized to the somatic region, (2) the initial segment spike is a low-threshold spike probably located at the axon hillock, (3) the somatodendritic spikes are long duration spikes that rapidly inactivate with depolarization, have a high threshold, and are localized to the dendritic regions. The action potential is then terminated by a long duration afterhyperpolarization. Our data further suggest that spike generation may be initiated by a slow depolarization at the soma triggering a spike in the low-threshold axon hillock which then spreads across the already-depolarized soma to trigger the dendritic spike. Based on the above findings, dopamine neurons can be compartmentalized electrophysiologically and morphologically into subcomponents, each associated with spikes and specific ionic currents. The high threshold dendritic component of the action potential demonstrates rapid inactivation with depolarization, and thus occurs over a rather narrow range of membrane polarization. This limited range of action potential generation may be important in control of dendritic dopamine release and/or modulation of electrical coupling between dopaminergic neurons.

Intracellular and extracellular electrophysiology of nigral dopaminergic neurons--3. Evidence for electrotonic coupling.

Using three independent in vivo methods, we have obtained evidence for electrotonic coupling between sets of rat zona compacta dopaminergic neurons: (1) Lucifer yellow injection into single dopamine neurons resulted in labeling of two to five dopamine neurons in 18 out of 33 injections. Similar injections into reticular formation or nigral reticulata cells did not demonstrate multiple labeling. (2) Intracellular recording revealed spontaneously occurring small (3-15 mV) fast potentials that often triggered action potentials in dopamine neurons when the membrane potential was close to firing threshold. These fast potentials had a firing rate and pattern similar to that reported previously for extracellularly recorded dopamine neurons. Fast potentials were activated antidromically from the caudate nucleus at a latency similar to that reported for dopamine neurons, followed high frequency antidromic stimulation at a constant latency, and collided with spontaneously occurring fast potentials. However, directly elicited action potentials would not collide reliably with antidromically activated fast potentials. Intracellular injection of depolarizing or hyperpolarizing current increased and decreased, respectively, the rate of occurrence of these potentials. The firing rate of fast potentials could be increased and decreased by the intravenous administration of dopamine antagonists and agonists, respectively. (3) Simultaneous extracellular recording from pairs of DA neurons revealed numerous instances of synchronized action potentials. This was observed more frequently following intravenous haloperidol administration. Sets of burst firing dopamine neurons recorded simultaneously consistently demonstrated a decrease in the interspike interval as the burst progressed; a phenomenon commonly reported in other electrically coupled systems. Electrical coupling has been suggested to be present in sets of identified nigrostriatal dopamine neurons. Electrical communication between these neurons could be involved in modulating burst firing and in synchronizing dopamine release.

Characterization of dopamine release in the rat medial prefrontal cortex as assessed by in vivo microdialysis: comparison to the striatum.

Using the technique of perfusion microdialysis combined with a small-bore liquid chromatography system we have measured the basal and drug-induced fluxes of extracellular dopamine in the medial prefrontal cortex of chloral hydrate-anesthetized rats and have compared our findings in the cortex to that observed in the striatum. The results were as follows. (1) At a flow rate of 2 microliter/min, the basal level of dopamine in the medial prefrontal cortex was 0.28 +/- 0.1 (n = 32) fmol/microliter perfusate, which was nearly an order of magnitude less than that obtained from the striatum. (2) alpha-Methyl-para-tyrosine (150 mg/kg i.v.) significantly decreased the extracellular levels of striatal and cortical dopamine. The magnitude and duration of the responses were similar in both regions. (3) Local perfusion with 30 mM K+ had a more profound effect on dopamine release in the striatum than in the medial prefrontal cortex. The K(+)-induced release in both regions was significantly attenuated in the absence of Ca2+. (4) The anxiogenic beta carboline FG 7142 (15 mg/kg, i.p.) enhanced the release of cortical dopamine by about 50% while it was without an effect in the striatum. (5) Amphetamine (1 mg/kg, i.v.) significantly elevated, while reserpine (5 mg/kg, i.p.) rapidly attenuated, the dopamine level in the medial prefrontal cortex. These studies demonstrate that perfusion microdialysis, in conjunction with small-bore liquid chromatography with electrochemical detection, can be used to measure the basal release of dopamine in the rat medial prefrontal cortex and that the dopamine release process in this region, as has been shown in the striatum, is sensitive to stimulation conditions and pharmacological manipulations.(ABSTRACT TRUNCATED AT 250 WORDS)

Opposite modulation of cortical N-methyl-D-aspartate receptor-mediated responses by low and high concentrations of dopamine.

To examine whether dopamine modulates cortical N-methyl-D-aspartate receptor-mediated glutamate transmission, whole-cell recordings were made from identified pyramidal cells located in layers V and VI of the medial prefrontal cortex of the rat using a slice preparation. In the presence of tetrodotoxin and the absence of Mg2+, a brief local application of N-methyl-D-aspartate evoked an inward current which was blocked by the N-methyl-D-aspartate antagonist dizocilpine maleate but not affected by the non-N-methyl-D-aspartate antagonist 2,3-dihydroxy-6-nitro-7-sulfamoyl-benzo(f)quinoxaline, suggesting that the observed current is mediated by N-methyl-D-aspartate receptors located on recorded cells. Bath application of dopamine produced opposite effects on the N-methyl-D-aspartate current depending on the concentrations of dopamine applied. At low concentrations (<50 microM), dopamine enhanced the N-methyl-D-aspartate current, whereas at higher concentrations, dopamine suppressed the current. The same concentrations of dopamine did not significantly affect the inward current induced by the non-N-methyl-D-aspartate agonist alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid. The enhancing effect of dopamine on the N-methyl-D-aspartate response was mimicked by the D1 agonist SKF38393 and blocked by the D1 antagonist SCH31966, whereas the suppressing effect was mimicked by the D2 agonist quinpirole and blocked by the D2 antagonist eticlopride. The above results suggest that dopamine at low concentrations acts preferentially on D1-like receptors to promote N-methyl-D-aspartate receptor-mediated transmission, while at high concentrations dopamine also activates D2-like receptors, leading to a suppression of the N-methyl-D-aspartate function. This differential modulation of N-methyl-D-aspartate function may have significant implications for understanding behaviors and disorders involving both cortical dopamine- and glutamate-mediated neurotransmission.

Evidence for the absence of impulse-regulating somatodendritic and synthesis-modulating nerve terminal autoreceptors on subpopulations of mesocortical dopamine neurons.

Electrophysiological and biochemical techniques were used to study midbrain dopamine systems. In the electrophysiological studies, projection areas of individual dopaminergic cells were identified by antidromic activation. Dopamine cells which innervate the piriform cortex and those that innervate the caudate nucleus demonstrated their usual dose-dependent inhibitory response to both the intravenous administration of the direct-acting dopamine agonist apomorphine and the microiontophoretic application of dopamine. In contrast, the firing rate of dopamine neurons which project to the prefrontal cortex and of those terminating in the cingulate cortex was not altered by either the intravenous administration of low to moderate doses of apomorphine or microiontophoretically applied dopamine. The mean basal discharge rate and degree of burst firing was also different between these subgroups of midbrain dopaminergic neurons. Mesoprefrontal and mesocingulate dopamine neurons had mean firing rates of 9.3 and 5.9 spikes/s respectively, and showed intense burst activity. Mesopiriform and nigrostriatal dopamine cells had discharge rates of 4.3 and 3.1 spikes/s and displayed only moderate bursting. The dopaminergic nature of those mesocortical neurons insensitive to apomorphine and dopamine was confirmed using combined intracellular recording and catecholamine histofluorescence techniques. Thus, after the intracellular injection of colchicine and subsequent processing for glyoxylic acid-induced histofluorescence, the injected cells could be identified by their brighter fluorescences compared to the surrounding, normally fluorescing, non-injected dopamine neurons. Using biochemical techniques, subgroups of midbrain dopaminergic systems were again found to differ. The administration of gamma-butyrolactone increased dopamine levels in all areas sampled (prefrontal, cingulate and piriform cortices as well as the caudate nucleus). However, although this effect was readily reversed in both the piriform cortex and caudate nucleus by pretreatment with apomorphine, this treatment had no effect on the increased dopamine levels observed in the prefrontal and cingulate cortices. In addition, the decline in dopamine levels after synthesis inhibition with alpha-methyltyrosine was significantly faster in the prefrontal and cingulate cortices relative to the caudate nucleus. The piriform cortex showed an intermediate decline which was not significantly different from that observed in any of the other regions.(ABSTRACT TRUNCATED AT 400 WORDS)

The treatment of tardive dyskinesia with baclofen.

Thirty-one psychiatric outpatients with tardive dyskinesia (TD) on neuroleptic medication were followed in a double-blind, randomized trial comparing baclofen (30-90 mg per day) to placebo. A repeated measures analysis of variance revealed no statistical difference between the baclofen-treated group and the placebo group for the total Abnormal Involuntary Movement Scale (AIMS) scores. There was a trend (P = 0.09) for an initial improvement, then a worsening of frequency counts across four visits. The authors attempt to explain this finding on the basis of information obtained from animal research.

Mechanisms of action of atypical antipsychotic drugs. Implications for novel therapeutic strategies for schizophrenia.

The mechanisms which contribute to the actions of atypical antipsychotic drugs, such as clozapine and the putative atypical agents remoxipride and raclopride, are reviewed. Examination of available preclinical and clinical data leads to two hypotheses concerning the mode of action of atypical antipsychotic drugs. The first hypothesis is that antagonism of the dopamine D2 receptor is both necessary and sufficient for the atypical profile, but that interaction with subtypes of the D2 receptor differentiates typical from atypical antipsychotic drugs. The second hypothesis has been previously advanced, and suggests that a relatively high ratio of serotonin 5-HT2:dopamine D2 receptor antagonism may subserve the atypical profile. It seems likely that the atypical antipsychotic drug profile may be achieved in more than one way.