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The contribution of endogenously released dopamine, GABA and its co-transmitters, substance P (SP) and neurokinin A (NKA), to the control of the evoked release of acetylcholine was investigated in vitro in the striosomes and the matrix of the rat striatum under various modalities of NMDA receptor stimulation (NMDA 50 microM or 1 mM without or with 10 microM D-serine). Sulpiride, bicuculline, SR140333 and SR48968, the antagonists of D(2), GABA A, NK(1) and NK(2) tachykinin receptors, respectively, were used for this purpose. (1) In both striatal compartments, the dopamine-mediated inhibitory regulation of the evoked release of acetylcholine only occurred when D-serine was co-applied with 50 microM or 1 mM NMDA. (2) In striosomes, the dopamine-dependent inhibitory effects of SP and NKA on the evoked release of acetylcholine only occurred when D-serine was co-applied with 50 microM or 1 mM NMDA. (3) A similar inhibitory regulation by NKA, but not SP, was found in the matrix when 1 mM NMDA was co-applied with D-serine. (4) In contrast, the dopamine-dependent facilitatory effect of GABA on the evoked release of acetylcholine did not require added D-serine and was more important with 1 mM than 50 microM NMDA. In the presence of D-serine, and depending on the NMDA concentration, the facilitatory regulation of GABA was reduced (matrix) or suppressed (striosomes). This latter effect was partially restored in the presence of SR48968. Therefore, the dopamine-dependent inhibitory effects of tachykinins on the evoked release of acetylcholine only occurred when NMDA receptors were stimulated in the presence of saturating concentrations of D-serine.

The N-methyl-D-aspartate-evoked release of [3H]acetylcholine previously formed from [3H]choline was estimated in striosome- (identified by [3H]naloxone binding) or matrix-enriched areas of the rat striatum using an in vitro microsuperfusion procedure. Experiments were performed in either the absence or the presence of dopaminergic and/or GABAergic receptor antagonists. Although the cell bodies of the cholinergic interneurons were mainly found in the matrix, in the absence of magnesium, N-methyl-D-aspartate (50 microM) stimulated the release of [3H]acetylcholine in both striatal compartments. These responses were blocked by either magnesium, dizocilpine maleate, 7-chlorokynurenate or tetrodotoxin. N-Methyl-D-aspartate responses were concentration-dependent, but the 1 mM N-methyl-D-aspartate response was higher in striosomes than in the matrix. The co-application of D-serine (10 microM) enhanced the 10 microM N-methyl-D-aspartate response in both compartments, but reduced those induced by 1 mM N-methyl-D-aspartate, this reduction being higher in striosomes. The blockade of dopaminergic transmission with the D2 and D1 dopaminergic receptor antagonists, (-)-sulpiride (1 microM) and SCH23390 (1 microM), was without effect on the 50 microM N-methyl-D-aspartate-evoked release of [3H]acetylcholine, but markedly enhanced the 1 mM N-methyl-D-aspartate+D-serine-evoked response in striosomes and to a lesser extent in the matrix. Disinhibitory responses of similar amplitude were observed not only in striosomes but also in the matrix when (-)-sulpiride was used alone, while SCH23390 alone enhanced the 1 mM N-methyl-D-aspartate+D-serine response only in striosomes and to a lower extent than (-)-sulpiride. These results indicate that D2 receptors are mainly involved in the inhibitory effect of dopamine on the 1 mM N-methyl-D-aspartate+D-serine-evoked release of [3H]acetylcholine. They also show that the stimulation of D1 receptors can either reduce (striosomes) or enhance (matrix) this response, since in the latter case the effect induced by the combined application of the D1 and D2 receptor antagonists was smaller than that observed with the D2 receptor antagonist alone. Indicating that released GABA facilitates N-methyl-D-aspartate responses, the blockade of GABAA receptors with bicuculline (5 microM) reduced the 50 microM N-methyl-D-aspartate-evoked release of [3H]acetylcholine in both striatal compartments and the 1 mM N-methyl-D-aspartate+D-serine response in the matrix. These effects result from an inhibition by GABA of the evoked release of dopamine, since the reducing effects of bicuculline on N-methyl-D-aspartate responses were not observed under the complete blockade of dopaminergic transmission by the D1 and D2 receptor antagonists. Further demonstrating a facilitatory role of GABA in the control of N-methyl-D-aspartate-evoked release of [3H]acetylcholine, in the presence of bicuculline, (-)-sulpiride and SCH23390 alone or in combination enhanced, in both compartments, the responses induced not only by 1 mM N-methyl-D-aspartate+D-serine, but also by 50 microM N-methyl-D-aspartate.

Using a new superfusion procedure in vitro, we have previously reported that the NMDA-evoked release of newly synthesized 3H-dopamine (DA) was higher in matrix- than in striosome-enriched areas of the rat striatum. In addition, GABAergic medium-sized spiny neurons were shown to be indirectly involved in this regulation. Since dynorphin and GABA are colocalized in a population of medium-sized spiny neurons, the role of dynorphin-containing neurons in the NMDA-evoked release of 3H-DA has been investigated using the same superfusion procedure on rat striatal slices. (1) The NMDA (50 microM, 25 min application)-evoked release of 3H-DA was increased in the presence of naloxone (1 microM, continuously delivered) in both striatal compartments, the overall response being more elevated in the striosome-enriched area. (2) The TTX (1 microM, continuously delivered)-resistant NMDA-evoked responses were also enhanced in the presence of naloxone, but in this case, the disinhibitory effects of naloxone were similar in striosome- and matrix-enriched areas. (3) The selective kappa-agonist U-50488 (1 microM) totally reversed the naloxone-disinhibitory effect on the NMDA-evoked response in the matrix-enriched area, but only partially in the striosome-enriched area. It also completely prevented the disinhibitory effect of naloxone on the TTX-resistant NMDA-evoked release of 3H-DA in both compartments. (4) The bicuculline (5 microM)- and naloxone (1 microM)-disinhibitory effects on the NMDA-evoked release of 3H-DA were additive in the matrix- but not in the striosome-enriched areas.(ABSTRACT TRUNCATED AT 250 WORDS)

Using an in vitro microsuperfusion device we have previously demonstrated that in the absence of magnesium, the N-methyl-D-aspartate-evoked release of [3H]dopamine (continuously synthesized from [3H]tyrosine) is more prominent in matrix- than in striosome-enriched areas of the rat striatum and that in the matrix, the response is partially tetrodotoxin-sensitive. Since the medium-sized GABAergic neurons are the main targets of the corticostriatal glutamatergic fibers, the involvement of local GABAergic regulation in the N-methyl-D-aspartate-evoked release of [3H]dopamine was investigated in both striatal compartments using the same experimental approach. Firstly, bicuculline alone (5 microM, 25-min application) was shown to enhance the release of [3H]dopamine similarly in both compartments revealing the existence of a tonic GABAergic control of the spontaneous release of [3H]dopamine. Secondly, the N-methyl-D-aspartate (50 microM, 25-min application)-evoked release of [3H]dopamine was markedly amplified in the presence of bicuculline (5 microM, continuous delivery). This effect being more important in striosome- than in matrix-enriched areas (5.5- and two-times the N-methyl-D-aspartate-evoked response observed in the absence of the GABAA antagonist, respectively). Thirdly, the tetrodotoxin (1 microM, continuous delivery)-resistant N-methyl-D-aspartate-evoked responses were also enhanced in the presence of bicuculline, but in this case, the amplification of the N-methyl-D-aspartate-evoked release of [3H]dopamine was less marked than in the absence of tetrodotoxin and identical in both compartments (about two-times the tetrodotoxin-resistant N-methyl-D-aspartate-evoked responses observed in the absence of bicuculline). Altogether, these results indicate that GABAergic neurons exert locally an important inhibitory regulation of the N-methyl-D-aspartate-evoked release of dopamine and that this effect is more prominent in the striosome-enriched area. Both tetrodotoxin-sensitive (striosome) and tetrodotoxin-resistant (striosome and matrix) processes intervene in this inhibitory GABAergic presynaptic regulation of dopamine release.

The release of [3H]gamma-aminobutyric acid ([3H]GABA) continuously formed from [3H]glutamine has been measured with a push-pull cannula implanted in the substantia nigra of the rat anesthetized with ketamine. Consistent with the high density of GABA terminals coming from both the striato-pallido-nigral afferents, and from GABA nigrofugal neurons, our results showed that a large amount of [3H]GABA was spontaneously released in the reticulata, about 4 times higher than in the compacta. In the absence of calcium the spontaneous [3H]GABA release was reduced (-30%), as well as the K(+)-induced release of [3H]GABA (-66%). Bicuculline (10(-4) M) did not affect the K(+)-evoked release of [3H]GABA, suggesting that autoreceptors on GABA afferent fibers are distinct from the GABAA subtype. Partial lesions of striato- and pallido-nigral GABA neurons with kainic acid (1.2 micrograms) decrease by 40% the glutamic acid decarboxylase (GAD) activity in the ipsilateral SN without decreasing the spontaneous release of [3H]GABA; even following extensive lesions with kainic acid (2.5 micrograms), GAD activity (-72%) and spontaneous [3H]GABA release (-83%) were not completely abolished. These results suggest that a non-negligible contribution of GABA nigral neurons accounts for the spontaneous GABA release measured in the substantia nigra. This is further supported by the decrease (-20%), and the increase (+40%) of [3H]GABA release produced by the local application of glycine (10(-6) M), and bicuculline (10(-4) M), which respectively, inhibits and activates the nigral neuron activity. The contribution of nigral GABA neurons to the amount of [3H]GABA release from the substantia nigra, is likely linked to their high spontaneous firing rate.

The release of gamma-[3H]aminobutyric acid ([3H]GABA) newly synthesized from [3H]glutamine was estimated in the superior colliculus of ketamine-anesthetized rats superfused via a push-pull cannula. A significant amount of [3H]GABA was spontaneously released in the superior colliculus (582 +/- 49 pCi/10 min). A major part of the large K(+)-evoked increase of the [3H]GABA release was Ca2+ dependent. When neuronal activity of the substantia nigra was enhanced by nigral application of K+ (30 mM) or bicuculline (10(-4) M), a persistent increase of the collicular [3H]GABA release was observed (60 and 80%, respectively). Conversely, when nigral activity was reduced by nigral application of GABA (10(-4) M) or superfusion with a Ca(2+)-free medium, a sustained decrease of the collicular [3H]GABA release was observed (-30 and -40%, respectively). Following the nigral application of a selective D2-receptor agonist. RU 24926 (10(-6) M), for 30 min in 6-hydroxydopamine-lesioned rats, a phasic increase (60%) of the collicular [3H]GABA release was detected. This effect could result from an activation of nigrocollicular GABAergic neurons by D2-receptor stimulation, because nigral activity and collicular release of [3H]GABA changed in a parallel direction.

Halothane-anaesthetized cats implanted with push-pull cannulae were used in this study. Amphetamine was applied in the pars reticulata or pars compacta of the substantia nigra in order to determine the role of dopamine released from distal or proximal dendrites of dopaminergic cells in the control of GABAergic transmission in the nucleus ventralis medialis of the thalamus. When applied for 30 min in either the pars reticulata or the pars compacta, amphetamine (10(-6) M) enhanced to a similar extent the local release of [3H]dopamine synthesized from [3H]tyrosine, these effects being seen mainly during the drug application. The amphetamine-evoked release of dopamine in the pars reticulata produced a long lasting reduction in the release of [3H]GABA synthesized from [3H]glutamine in the nucleus ventralis medialis as well as in the paralamellar zone of the nucleus ventralis lateralis. Opposite effects were observed when amphetamine (10(-6) M) was applied in the pars compacta. In complementary experiments, single unit recordings were made in the intermediate part of the pars reticulata, some of the cells being identified by antidromic activation from the nucleus ventralis medialis. Whether applied in the pars reticulata or pars compacta, amphetamine (10(-6) M, 10 min) evoked a reversible decrease in the firing rate of most recorded cells whether or not they were identified as projecting to the nucleus ventralis medialis. Therefore, the decreased release of [3H]GABA in the nucleus ventralis medialis seen following application of amphetamine in the pars reticulata of the substantia nigra could result from an inhibition of nigrothalamic GABAergic neurons. Since the nucleus ventralis medialis is also innervated by GABAergic neurons originating in the entopeduncular nucleus, single unit recordings were made from cells in this nucleus during the application of amphetamine (10(-6) M, 10 min) into the pars compacta of the substantia nigra, some of which were identified antidromically as projecting to the nucleus ventralis medialis. Most cells identified or not were found to be activated during this treatment. These results suggested that the increased release of [3H]GABA seen in the nucleus ventralis medialis following application of amphetamine in the pars compacta of the substantia nigra might be linked to the enhanced firing rate of entopeduncular-thalamic GABAergic neurons.

Push-pull cannulae were implanted in both caudate nuclei and both substantiae nigrae (SN) of halothane-anesthetized cats and the release of [3H]GABA, continuously synthesized from [3H]glutamine, was measured in these structures during 4 h of superfusion. In some experiments, multi-unit neuronal activity was recorded at the tip of the nigral push-pull cannulae, using a bipolar electrode. Two hours after the onset of superfusion with [3H]glutamine, 10(-6)M of muscimol was added (for 1 h) in the superfusion medium delivered to one SN. This treatment increased locally the release of [3H]GABA and enhanced the neuronal activity of the nigral cells in the zona reticulata. An increased release of [3H]GABA was also observed in the contralateral SN, in association with an inhibition of the activity of the zona reticulata cells. The unilateral nigral application of muscimol also induced asymmetric changes in the release of [3H]GABA in both caudate nuclei, since [3H]GABA release was increased ipsilaterally and reduced on the contralateral side. The present findings are considered in relation to possible GABAergic neuronal populations affected by this local pharmacological treatment.

In halothane anaesthetized cats, a push-pull cannula was implanted into the right caudate nucleus (CN) and in each substantia nigra (SN). The release of [3H]GABA continuously formed from [3H]glutamine was estimated in each structure. Acetylcholine (ACh, 5 x 10(-5) M) added in presence of eserine (5 x 10(-5) M) for 50 min in the right caudate nucleus 2 h after the onset of superfusion with [3H]glutamine, stimulated the [3H]GABA release locally. The effect was biphasic when ACh application was made in the median two-thirds of the structure and it was monophasic and transient when the ACh application was restricted to the lateral part. ACh application in the right caudate nucleus also induced changes in [3H]GABA released in the anterior (pars reticulata) and posterior (pars compacta) parts of both SN. While [3H]GABA release was enhanced in the ipsilateral anterior SN, it was reduced in the contralateral anterior SN. Respective opposite effects were observed in the posterior parts of the ipsi- and contralateral SN. These bilateral asymmetrical changes in [3H]GABA release were not dependent on the site of ACh application in the right caudate nucleus. These results indicate that the facilitation of cholinergic transmission in one caudate nucleus influences in an opposite way the striato-nigral GABA neurones on both sides of the brain.

The release of [3H] gamma-aminobutyrate (GABA) neosynthesized from [3H]glutamine was estimated in one substantia nigra and in the ipsilateral thalamus of halothane-anesthetized cats by perfusing a [3H]glutamine-enriched physiological medium through a push-pull cannula implanted in the two structures under investigation. After two hours of superfusion, muscimol (10(-6)M) was delivered through the nigral push-pull cannula for 50-60 min and local- and distal-evoked changes of [3H]GABA release were analyzed. In some experiments, changes of global neuronal activity induced by muscimol application were recorded in different thalamic nuclei, using a bipolar electrode. In a few of the above experiments, biochemical and electrophysiological determinations were simultaneously performed in the substantia nigra and the thalamus. The nigral application of muscimol (10(-6)M) induced locally an activation of the substantia nigra reticulata cells, as well as an increase in release of [3H]GABA. Distally, in the thalamus, two types of biochemical and electrophysiological responses were observed according to the localization of the tip of the push-pull cannula or the electrode. (1) An increased release of [3H]GABA and a depression of the global multi-unit cellular activity were obtained in the ventralis medialis-ventralis lateralis, the centralis lateralis and the paracentralis nuclei. These effects could reflect an activation of the GABAergic nigrothalamic neurons projecting to these different thalamic nuclei. (2) In contrast, in the medialis dorsalis paralamellar zone adjacent to the intralaminar nuclei of the thalamus, a decrease of [3H]GABA release and an activation of the multi-unit activity were obtained. These latter results may suggest either a polysynaptic response or the non-GABAergic nature of the nigrothalamic neurons afferent to the medialis dorsalis paralamellar zone.

Halothane anesthetized cats were implanted with push--pull cannulae to study the release of [3H]GABA continuously formed from [3H]glutamine in the substantia nigra (SN) and in the pallido-entopeduncular nuclei (PEP). A spontaneous release of [3H]GABA was observed from both structures and it reached a steady state level 1 h after the beginning of the superfusion with [3H]glutamine. In cats implanted with two push--pull cannulae, the local application of potassium (47 mM) in the PEP stimulated the release of [3H]GABA from the ipsilateral SN. In cats implanted with 4 push--pull cannulae, the unilateral 10 min electrical stimulation of the caudate nucleus evoked the release of [3H]GABA not only from the ipsilateral SN, but also in most cases from the contralateral structure. This stimulus also enhanced the release of [3H]GABA from PEP but the effects were mainly observed in the medio-caudal part of the ipsilateral PEP and in the latero-rostral part of the contralateral structure. In all cases, the changes in [3H]GABA release were observed during and after the electrical stimulation. The ipsilateral effects can be attributed to the direct activation of the caudato-PEP or caudato-SN GABAergic neurons. A polysynaptic neuronal loop must be involved in the symmetric contralateral effects.

The contribution of motor and intralaminar thalamic nuclei to the changes of [3H]GABA release evoked in both caudate nuclei (CN) and both substantia nigra (SN) by a unilateral nigral application of muscimol (10(-6) M) was investigated on halothane-anaesthetized cats. Acute lesions were performed on one side of the thalamus at the level of either the ventralis medialis and ventralis lateralis (motor nuclei) or the centralis lateralis and paralamellar zone of the medialis dorsalis (intralaminar nuclei). The release of [3H]GABA neosynthesized from [3H]glutamine was measured by perfusing continuously a [3H]glutamine-enriched physiological medium through a push-pull cannula implanted in the 4 structures under investigation. After two hours of superfusion, muscimol (10(-6) M) was delivered for 60 min through the nigral push-pull cannula implanted ipsilaterally to the thalamic lesion. Evoked changes of [3H]GABA release were analyzed either in motor or intralaminar nuclei lesioned cats and compared to those observed in intact animals. Whatever the localization of the thalamic lesions was, an increased release of [3H]GABA was elicited locally in the SN and distally in the ipsilateral CN as in intact animals, suggesting that the responses induced ipsilaterally did not require nigro-thalamic pathways. On the contrary, in the contralateral CN changes of [3H]GABA release evoked by the nigral muscimol application were reversed by both types of thalamic lesion. Instead of a decreased release of [3H]GABA observed in intact cats, an increased release of [3H]GABA was detected in lesioned animals. In the contralateral SN, the response was reversed only after the intralaminar nuclei lesion. In this situation nigral muscimol application induced a decreased release of [3H]GABA in contrast to the enhanced release observed in intact and motor thalamic lesioned cats. The parallel increased release of [3H]GABA observed in the contralateral CN and SN in motor thalamic nuclei lesioned cats suggests an activation of the striatonigral cells by the nigral muscimol treatment. The asymmetrical changes of [3H]GABA release measured in the contralateral CN and SN in intact and intralaminar nuclei lesioned cats could indicate a presynaptic modulation of the [3H]GABA release acting either at the CN or the SN levels. The possible pathways involved in the interhemispheric transfer of information originating from one SN to the contralateral basal ganglia components are also discussed.