Axonal transport of proteins and glycoproteins in the rat nigro-striatal pathway and the effects of 6-hydroxydopamine

Following stereotaxic injection of [35S]methionine into the substantia nigra of adult rats, there was rapid local incorporation of radioactivity into acid-insoluble material. Incorporation peaked by 4 h and then decreased. In contrast, acid-precipitable radioactivity in the corpus striatum (the major projection site of the substantia nigra) rose markedly between 1 and 8 h followed by a plateau period and another even more marked increase between 24 h and 6 days. Experiments involving injection of [3H]fucose gave similar results except that most of the acid-precipitable radioactivity in the striatum appeared in an early wave. In each case radioactivity in the contralateral striatum was less than 11% of that on the ipsilateral side. Stereotaxic injection of colchicine (20 microgram) into the nigrostriatal pathway (within the median forebrain bundle) blocked transport of [35S]protein and [3H]glycoprotein by 90% and 50%, respectively. In animals treated with 6-hydroxydopamine (6-OHDA; treated neonatally or as adults) the accumulation of striatal [35S]protein was reduced to 7 to 26% of control levels; striatal [3H]glycoprotein was also reduced, but not as much (29% to 73% of control). In control experiments, [3H]DOPA wa injected into the substantia nigra, and [3H]dopamine was measured in corpus striatum; 6-OHDA treatment reduced the amounts of striatal [3H]dopamine recovered to 3% of control values. The failure of colchicine or 6-OHDA to block transport of incorporated fucose as effectively as the transport of incorporated methionine is possible due to greater diffusion of fucose away from the injection site to non-dopaminergic nuclei projecting to the striatum. The molecular weight distribution of radioactive proteins at the substantia nigra and corpus striatum was analyzed by polyacrylamide gel electrophoresis. For both [35S]methionine and [3H]fucose, the gel electrophoretic pattern of radioactive proteins in the injection site (substantia nigra) was complex and did not change greatly between 2 h and 6 days. At the projection site (striatum) the electrophoretic distribution pattern was initially different from that of the substantia nigra, and changed markedly over the course of several days. In 6-OHDA-treated animals (treated neonatally or as adults), the bulk of proteins transported in nigro-striatal non-dopaminergic neurons appears to be very similar to that transported in the intact pathway in control rats. However, in striata of 6-OHDA-treated animals, a consistent reduction in striatal 35S- and 3H-radioactivitiy was observed in proteins with molecular weight from about 67,000 to 77,000. Assuming that the 6-OHDA treatment did not substantially affect the non-dopaminergic neurons, we interpret this to mean that some of the proteins in this molecular weight range are transported primarily by dopaminergic neurons.     

Changes in 3H-substance P receptor binding in the rat brain after kainic acid lesion of the corpus striatum

Previous studies have indicated that the substantia nigra contains the highest concentration of substance P-like immunoreactivity (SPLI) in the brain. Paradoxically, it also appears to contain one of the lowest concentrations of substance P receptors in the brain. One possibility is that the massive amount of SPLI blocks the binding of the radioligand to the substance P receptor and/or "down-regulates" the number of substance P receptors present in this structure. Since greater than 95% of the SPLI within the substantia nigra originates from the corpus striatum, we have lesioned this area and measured the changes in substance P receptor concentration in the substantia nigra and other corpus striatal projection areas. A semiquantitative autoradiographic technique for measuring the binding of 3H-substance P to substance P receptors was used in conjunction with tritium-sensitive film. 3H-substance P binding was measured in both the corpus striatum and its projection areas after kainic acid lesion of the corpus striatum. At either 4 or 21 d after the lesion there was approximately a 90% loss of substance P receptors in the rostral striatum, a 74% loss in the globus pallidus, a 57% increase in receptor number in lamina I and II of the ipsilateral somatosensory cortex, and no apparent change in the number of receptors in the substantia nigra pars reticulata, superior colliculus, and central gray. These findings suggest that the low concentration of substance P receptors found within the substantia nigra is not due the massive SPLI innervation, since removal of greater than 95% of the SPLI had no measurable effect on the concentration of substance P receptors.(ABSTRACT TRUNCATED AT 250 WORDS)     

In vivo biosynthesis of [35S]- and [3H]substance P in the striatum of the rat and their axonal transport to the substantia nigra

The biosynthesis and axonal transport of the neuropeptide substance P (SP) in the striatonigral tract of the rat was examined using an in vivo radiolabeling of the rostral corpus striatum and a series of high performance liquid chromatography (HPLC) steps for the purification of radiolabeled SP. The corpus striatum of unrestrained rats was continuously infused via indwelling cannulae for 16 hr with [35S]methionine or a mix of [3H]leucine and [3H]proline. Radiolabeled SP was acid extracted from discrete regions of this striatonigral SP projection--corpus striatum (SP-immunopositive cell bodies), ansa lenticularis (striatonigral SP axons), and substantia nigra (striatonigral SP terminals)--and was purified to a constant specific activity by sequential HPLC. The radiochemical purity of SP was verified by chemical derivative formation (SP-Met11-sulfoxide) and further HPLC. The in vivo labeling procedure resulted in a high level of incorporation of the amino acids into tissue protein and peptide pools. [35S]SP and [3H]SP were positively identified in all three regions of this peptidergic projection. The amount of [35S]SP harvested in each region accounted for 0.0015%, 0.003%, and 0.071% of the total tissue 35S present in the striatum, striatonigral fibers, and substantia nigra, respectively. The amount of [3H]SP harvested in each region accounted for 0.0025%, 0.011%, and 0.27% of the total tissue 3H present in the three regions, respectively. The amount of radiolabeled SP in the striatonigral regions for both isotopic infusion studies was highly correlated with the immunoassayable SP content in those regions, suggesting rapid equilibration of de novo biosynthesized SP with striatonigral tissue pools of SP. Tritium autoradiography of the striatonigral projection after 3H-amino acid infusion provided further support for the specificity of the radiolabeling procedures. Heavily labeled fibers were seen leaving the striatal infusion site caudally, forming a distinct fiber bundle. This bundle projected caudoventrally and formed a dense terminal plexus primarily within the reticulata portion of the substantia nigra. These results demonstrate that SP biosynthesis in the corpus striatum and its transport to the substantia nigra can be studied in discrete striatonigral regions obtained from individual unrestrained rats. This preparation should allow for studies on the dynamics of SP biosynthesis, axonal transport, and turnover in the striatonigral projection.     

In vivo labelling and axonal transport of monoamine oxidase in the rat basal ganglia using radioactive pargyline

Summary The enzyme monoamine oxidase was labelled in the rat striatum or substantia nigra with locally injected radioactive pargyline. The binding was prevented by a pretreatment with non-radioactive pargyline, or with a combination of clorgyline and deprenyl. Most of the MAO labelled with³H-pargyline was of the B-type, but also some MAO-A was labelled, as shown in rats pretreated with clorgyline or deprenyl separately.Seven days after the injection of (³H)-pargyline into the striatum a significant labelling was observed in the substantia nigra. This labelling was clorgyline sensitive, indicating type A MAO, and was not present when striatal neurons were destroyed with kainic acid. Labelling of the striatum following³H-pargyline injection into the substantia nigra was also less in kainate intoxicated striata. Damage of nigral dopamine neurons with 6-hydroxydopamine did not influence the distribution of the label.Thus by using³H-pargyline, specific labelling and axonal transport of type A MAO in striatal neurons projecting to the substantia nigra was demonstrated.     

Chronic haloperidol and clozapine differentially affect dynorphin peptides and substance P in basal ganglia of the rat

The effect of chronic neuroleptic treatment, using haloperidol or clozapine, on immunoreactive dynorphin peptide and substance P levels in basal ganglia of rats was examined. The drugs were administered i.p. in daily doses for 10 days (haloperidol 1 mg/kg and clozapine 10 mg/kg). Dynorphin A, dynorphin B and substance P were measured in substantia nigra, striatum, globus pallidus and hypothalamus using specific radioimmunoassays. The most prominent effects were observed with with clozapine which increased levels of all measured peptides in substantia nigra. Haloperidol only affected nigral substance P levels which declined, while nigral dynorphin peptide levels remained unchanged. In striatum, haloperidol slightly reduced dynorphin peptides while substance P was unaffected. Clozapine increased striatal substance P but the dynorphin peptides were not affected. Minor changes in dynorphin peptides found in globus pallidus and hypothalamus were not statistically reliable. Substance P was not changed in these structures after either of the two drugs. High molecular weight fragments (greater than or equal to 5,000) from the dynorphin precursor, proenkephalin B, were measured in substantia nigra and striatum using trypsin digestion and subsequent analysis of generated Leu-enkephalin-Arg6. These high molecular weight fragments were found to be affected in the same manner as the dynorphin peptides. This study indicates that the two types of neuroleptic drugs have different modes of interaction on peptide systems in basal ganglia of rats. Dynorphin peptides and substance P were also differentially affected.     

Immunohistochemical demonstration of differential substance P-, met-enkephalin-, and glutamic-acid-decarboxylase-containing cell body and axon distributions in the corpus striatum of the cat

The immunohistochemical localization of neuronal cell bodies and axons reactive for substance P (SP) and methionine-enkephalin (ME) was investigated in the corpus striatum of the adult cat brain and compared with that of glutamate decarboxylase (GAD), synthetic enzyme for gamma-aminobutyric acid. Striatal cell bodies reactive for ME could be identified only in colchicine treated cats, are medium size, ovoid striatal cells, and are found in large numbers in a more or less even distribution throughout the caudate nucleus, putamen, and nucleus accumbens. The striatal region most densely occupied by ME-immunoreactive cells is the ventral and central part of the caudate head. Modest numbers of larger ME-reactive neurons are dispersed throughout the entopeduncular nucleus and the pars reticulata of the substantia nigra. Striatal cells of medium size reactive for SP could be identified, with or without colchicine, in largest numbers in the medial half of the caudal three-fourths of the putamen and in clusters of irregular size and shape in the head of the caudate nucleus. Cells reactive for SP are also common in layer II and the islands of Calleja of the olfactory tubercle. We could not reliably visualize GAD-positive cell bodies in the striatum, even with colchicine treatment; however, they could be seen readily in all pallidal structures such as the globus pallidus, ventral pallidum, entopeduncular nucleus, and substantia nigra. Axons reactive for ME are found mainly in the globus pallidus where they form a dense and even network throughout the nucleus. The globus pallidus is almost devoid of SP reactivity except near its extreme caudal pole. Conversely, SP-immunoreactive axons form dense meshworks in the entopeduncular nucleus and substantia nigra where ME immunoreactivity is minimal. Fewer, but still ample numbers, of SP-reactive axons are present also in the ventral tegmental and retrorubral areas of the midbrain tegmentum and in the ventral pallidum of the basal forebrain, but only sparse ME-reactive axons are present in these areas. This differential distribution of SP- and ME-containing axons in the pallidal and nigral structures stands in contrast to the relatively homogeneous and dense distribution of GAD-containing axons throughout the dorsal and ventral pallidum, entopeduncular nucleus, and substantia nigra.(ABSTRACT TRUNCATED AT 400 WORDS)     

Loss of substance P and Enkephalin immunoreactivity in the human substantia nigra after striato-pallidal infarction

Post-mortem neuropathological material from 3 patients with striato-pallidal infarction provided the first immunohistochemical evidence for substance P- and enkephalin-containing nerve fibre projections from the striato-pallidum to the substantia nigra in the human. The nigra corresponding to the normal side showed abundant substance P and enkephalin immunoreactivity whose patterns of immunostaining were notably similar. In contrast, the substantia nigra ipsilateral to the striato-pallidal infarction showed a decrease in substance P and enkephalin immunoreactivity which was proportional to the extent of the infarction. This suggests that much of the substance P and enkephalin immunoreactivity in the nigra is present in nerve fibres projecting from the striato-pallidum. Furthermore, the similar distribution of remaining substance P and enkephalin immunoreactivity in corresponding areas of the nigra of the infarcted side indicates that the origins and/or projections of nerve fibres containing these two neuropeptides may be closely approximated anatomically. Depletion of substance P immunoreactivity in the pars reticulata of the substantia nigra in a fourth patient with anterior striatal infarction suggests a topographic projection for substance P immunoreactive fibres from the striatum. There is some evidence in one patient with cognitive and behavioural abnormalities to support the suggestion that the basal ganglia may be involved in non-motor functions.     

Direct demonstration of interactions between substance P immunoreactive terminals and tyrosine hydroxylase immunoreactive neurons in the substantia nigra of the rat: an ultrastructural study

The results of many anatomical, physiological, and pharmacological studies suggest that substance P-containing neurons of the striatum project to the substantia nigra, and that substance P influences the activity of dopaminergic nigrostriatal neurons. The purpose of the present ultrastructural study was to employ dual immunocytochemical labeling to determine the morphological basis for the observed actions of substance P on nigral dopaminergic neurons. Substance P-like and tyrosine hydroxylase-like immunoreactivities were localized simultaneously at the ultrastructural level in the substantia nigra of the rat. A double label method was utilized which relied on a combination of the peroxidase-antiperoxidase method (Sternberger, 1979) for substance P, and immunogold or silver enhanced immunogold labeling for tyrosine hydroxylase. The present results indicate that tyrosine hydroxylase immunoreactive (THLI) dendrites in the substantia nigra receive synaptic input from terminals exhibiting substance P-like immunoreactivity. These findings support the idea that substance P is a major neurotransmitter in the striatonigral loop, and suggest that striatal substance P neurons act directly upon nigral dopaminergic cells.     

Comparative distribution of mRNAs for glutamic acid decarboxylase, tyrosine hydroxylase, and tachykinins in the basal ganglia: an in situ hybridization study in the rodent brain

Neurotransmitter-related messenger RNAs were detected by in situ hybridization in sections of rat and mouse brains by using 35S-radiolabelled RNA probes transcribed from cDNAs cloned in SP6 promoter-containing vectors. The distribution of messenger RNAs for glutamic acid decarboxylase, tachykinins (substance P and K), and tyrosine hydroxylase was examined in the striatum, pallidum, and substantia nigra. Dense clusters of silver grains were observed with the RNA probe complementary of the cellular messenger RNA for glutamic acid decarboxylase (antisense RNA) over most large neurons in the substantia nigra pars reticulata and medium-sized to large neurons in all pallidal subdivisions. A few very densely and numerous lightly labelled medium-sized neurons were present in the striatum. Among the areas examined, only the striatum contained neurons labelled with the antisense tachykinin RNA. Most of these neurons were of medium size, and a few were large. With the antisense tyrosine hydroxylase RNA, silver grains were found over neurons of the substantia nigra pars compacta and adjacent A10 and A8 dopaminergic cell groups. No signal was observed with RNAs identical to the cellular messenger RNA for glutamic acid decarboxylase or tachykinin (sense RNA). These results show a good correlation with immunohistochemical studies, suggesting that documented differences in the distribution and the level of glutamic acid decarboxylase, tyrosine hydroxylase, and substance P immunoreactivities in neurons of the basal ganglia are related to differences in the level of expression of the corresponding genes rather than to translation accessibility, stability, or transport of the gene products.     

Substance P-dopamine relationship in the rat substantia nigra: a light and electron microscopy study of double immunocytochemically labeled materials

The synaptic relationships between substance P-containing terminals and dopaminergic neurons (immunoreactive for tyrosine hydroxylase) in the substantia nigra were studied at both light and electron microscopic levels using a pre-embedding double-labeling immunocytochemical method. Many substance P-containing terminals were found to form synapses directly with dendrites and somata of nigral dopaminergic neurons. Since most of the substance P-containing axon terminals arise from the striatum, this result suggests that striatal substance P neurons can have monosynaptic influence on nigral dopaminergic neurons.     

Localization of striatal and nigral tachykinin receptors in the rat

The effects of lesioning mesostriatal dopamine projections or striatal neurons on tachykinin binding in the basal ganglia were assessed in the rat. 6-Hydroxydopamine lesions of the medial forebrain bundle destroyed striatal dopamine terminals as assessed by [³H]mazindol autoradiography, but did not significantly affect the binding of NK-1 ([³H][Sar⁹, Met(O₂)¹¹]substance P) or NK-3 ([³H]senktide) tachykinin ligands in the striatum. 6-Hydroxydopamine lesions significantly reduced NK-3 binding in the substantia nigra pars compacta, but not the ventral tegmental area. In contrast, striatal quinolinic acid lesions reduced both NK-1 and NK-3 binding in the striatum, but failed to affect NK-3 binding in the substantia nigra. These findings suggest that both NK-1 and NK-3 receptors within the striatum are predominantly post-synaptic with respect to dopamine neurons, whereas nigral NK-3 receptors are located on dopaminergic neurons.     

Somatostatin is increased in the basal ganglia in Huntington disease

Huntington disease (HD) is an autosomal dominant hereditary disorder characterized by premature cell death, predominantly in the neostriatum. Decreased concentrations of several neurotransmitters and neuropeptides have been reported in the basal ganglia in Huntington disease. We now report that concentrations of radioimmunoassayable somatostatin are increased in extracts of the caudate (mean ± standard error of the mean, ng/gm net weight; 247± 24 versus 85 ± 11), putamen (275 ± 48 versus 74 ± 11), external globus pallidus (100 ± 10 versus 27 ±6), and internal globus pallidus (108± 21 versus 21 ± 8) in the disease. The concentrations of immunoreactive substance P measured in the same extracts were markedly reduced in caudate (mean ± standard error of the mean, pmol/gm wet wight; 25 ± 3 versus 109 ± 20), putamen (28 ± 7 versus 88 ± 28), external globus pallidus (39 ± 9 versus 196 ± 62), and internal globus pallidus (60 ± 17 versus 263 ± 39), as well as in both subdivisions of the substantia nigra. Gel permeation chromatography and high-performance liquid chromatography showed radioimmunoassayable somatostatin to include peptides with physicochemical properties of the tetradecapeptide somatostatin and larger substances, including somatostatin-28-like material. A single peak of immunoreactive substance P corresponding to synthetic substance P was found by high performance liquid chromatography. These results suggest that immunoassayable somatostatin-containing neuronal elements in the neostriatum and globus pallidus in Huntington disease are affected differentially by the disease process from neurons that contain immunoreactive substance P.     

Neuropeptides in striato-nigral pathways

Summary The existence of a neuronal pathway containing Leu-enkephalin and connecting the neostriatum with the globus pallidus has been confirmed combining immunohistochemistry with microinjections of neurotoxic agents (kainic acid, colchicine) and discrete knife lesions.The presence of substance P in nerve terminals of the substantia nigra was demonstrated by the application of a monoclonal antibody against this peptide. Electron microscopic studies revealed immunoreactive sites for substance P in nerve terminals establishing symmetric and asymmetric synapses, mainly over dendritic profiles.The possible peptide-containing neuronal pathways in the nigro-striatal system are discussed.     

Zinc transport from the striatum and substantia nigra

⁶⁵Zn was unilaterally injected into the striatum or substantia nigra of rats to see the transport of intracerebral zinc (Zn). In the case of intrastriatal injection, ⁶⁵Zn was densely distributed in the ipsilateral medial forebrain bundle (MFB) and the substantia nigra. On unilateral colchicine injection into the MFB, ⁶⁵Zn distribution in the ipsilateral substantia nigra decreased significantly compared to that of the contralateral one after ⁶⁵Zn injection into the bilateral striata. These results suggest the presence of axonal transport of ⁶⁵Zn taken up by striatonigral γ-aminobutyric acid (GABA)-ergic and/or nigro-striatal dopaminergic neurons. On the other hand, in the case of intranigral injection, ⁶⁵Zn was distributed in the ipsilateral MFB, striatum, globus pallidus, pontine reticular nuclei, and pontine nuclei. ⁶⁵Zn distribution in the pons 1 day after intranigral injection was very similar to that 6 days after intrastriatal injection, suggesting that, in the case of intrastriatal injection of ⁶⁵Zn, nigral ⁶⁵Zn, which was transported anterogradely and/or retrogradely from the striatum, was transported to the postsynaptic neurons through the synapse and then transported to the pons.     

Effect of the normal nerve impulse flow on the synthesis and utilization of GABA in the rat substantia nigra

Summary The concentrations of GABA, glutamate, serine, glutamine, threonine, glycine and taurine in the substantia nigra and in the corpus striatum of the rat were determined electrochemically following condensation with o-phthalaldehyde--mercaptoethanol and reverse-phase, high performance liquid chromatography.After a frontal hemisection at the level of the caudal hypothalamus, the GABA concentration in the substantia nigra on the operated side decreased to about 20 per cent of the normal value in 4 days, in all probability caused by degeneration of the nerve terminals of the striato-nigral GABA neurons. The concentrations of taurine in the substantia nigra and of GABA in the corpus striatum were initially lowered and later elevated following this lesion. The concentration of glutamate in the substantia nigra was lower on the sectioned side and higher on the intact side at 14 days as compared to 4 hours after a hemisection.Following an acute hemisection, the GABA transaminase inhibitor-acetylenic GABA increased the concentration of GABA by 36 % and 79 % in the substantia nigra on the sectioned and intact side, respectively. The glutamate decarboxylase inhibitors 4-deoxypyridoxine and isoniazid lowered the concentration of GABA in the substantia nigra by about 50% on both the sectioned and intact side. The results indicate that the synthesis, but not the utilization of GABA in the substantia nigra is dependent on the normal nerve impulse flow.The concentration of glutamine was changed in directions contrary to that of GABA following a chronic hemisection or treatment with-acetylenic GABA, in agreement with the suggestion that glutamine is a precursor of the GABA transmitter pool.     

Manganese Transport in the Neural Circuit of Rat CNS

To study manganese (Mn) transport in the neural circuit of rat CNS, brain isotope distribution after ⁵⁴Mn injection into the brain was analyzed by autoradiography. One day after ⁵⁴MnCl₂ injection into the striatum, ⁵⁴Mn was highly distributed in the ipsilateral thalamus, hypothalamus, and substantia nigra. When ⁵⁴MnCl₂ was bilaterally injected into the striata after unilateral treatment with colchicine or vehicle into the medial forebrain bundle, ⁵⁴Mn was distributed in both sides of the substantia nigra of vehicle-treated rats. On the other hand, unilateral colchicine treatment caused a decrease of ⁵⁴Mn distribution in the ipsilateral substantia nigra, suggesting that Mn is subjected to axonal transport in the striatonigra and/or nigrostriatal pathways. In the case of unilateral injection of ⁵⁴MnCl₂ into the olfactory bulb, ⁵⁴Mn was distributed in the ipsilateral piriform, amygdaloid areas (the primary olfactory cortex), and entorhinal area (the secondary olfactory cortex). These results suggest that Mn is subject to widespread axonal transport in the neural circuits. Moreover, Mn may be taken up by the piriform neurons (the third olfactory neuron) after release from the secondary olfactory neuron terminals and transported to the entorhinal area.     

Immunoreactive substance P in the substantia nigra of the monkey: light and electron microscopic localization

The microscopic localization of immunoreactive substance P in the monkey substantia nigra was investigated by the peroxidase antiperoxidase method. Immunoreactive substance P was identified in the substantia nigra pars compacta and pars reticulata in numerous myelinated and unmyelinated fibers, and within axon terminals which contained intensely labeled granular vesicles and formed predominantly axodendritic synapses. These morphologic results are consistent with known excitatory effects of SP on nigral neurons.     

Lack of effects of apomorphine,haloperidol and clozapine on the synthesis and utilization of brain GABA

Summary The synthesis of GABA was studied as the accumulation of GABA following inhibition of the GABA--ketoglutaric acid aminotransferase by-acetylenic GABA (GAG) or-vinyl GABA. The disappearance of GABA was studied by means of inhibition of glutamate decarboxylase by 4-deoxypyridoxine.Systemic administration of the dopamine receptor agonist apomorphine did not change the accumulation or the disappearance of GABA in the substantia nigra and the corpus striatum of the rat. However, a very high dose of apomorphine somewhat increased the GAG-induced GABA accumulation in the corpus striatum. The dopamine receptor antagonists haloperidol and clozapine did not modify the accumulation and disappearance of GABA in the two structures, except for a slight decrease in the GAG-induced GABA accumulation in the substantia nigra.Following an acute hemisection, the neural connections between the substantia nigra and the corpus striatum are interrupted on one side of the brain. Apomorphine did not influence the accumulation or the disappearance of GABA in the substantia nigra and in the corpus striatum on the sectioned side of the brain. These results indicate that the synthesis and the utilization of GABA in the two structures studied are not influenced to any greater extent by changes in dopamine receptor activity.     

Effects of KCl-induced depolarization on the GABA concentration in the corpus striatum and in the substantia nigra

Summary The effects of potassium ion depolarization on the concentration, the synthesis and the utilization of GABA as well as the effects on the concentrations of glutamine, dopamine (DA) and DOPAC in the substantia nigra and in the corpus striatum of the rat were investigated.An intranigral KCl injection did not influence the GABA concentration or the synthesis and utilization of GABA in the substantia nigra. Following an intrastriatal KCl injection the GABA concentration increased and the glutamine concentration decreased in the corpus striatum. A marked increase in the DA concentration in the corpus striatum was seen following an intranigral KCl injection, whereas the DA concentration decreased and the DOPAC concentration increased following an intrastriatal KCl injection. It is concluded that the GABA in the substantia nigra and in the corpus striatum is affected differently by potassium ions, perhaps due to the different cellular localization of GABA in the two structures. The increase of GABA in the corpus striatum was accompanied by a decreased glutamine concentration indicating that glutamine might be a precursor of GABA.