Characteristics of dopamine and GABA transport in primary cultures of astroglial cells

Primary astroglial cultures were grown from newborn rat cerebral hemispheres. Cultures, aged 7 and 14 days, were analysed with respect to their capacity to accumulate radioactive GABA and dopamine. Concentrative high-affinity uptake, showing Na+-dependence was observed for GABA where Km and Vmax were not significantly altered with age of the culture. No Na+-dependent high-affinity transport of dopamine was observed. The addition of 0.1 mM dibutyryl-cyclic-3'-5'-adenosinemonophosphate (dB-cAMP) after removal of fetal calf-serum reduced the Vmax as well as the Km of 3H-GABA uptake. The ionic dependence of 3H-GABA and 3H-dopamine was not significantly altered following this treatment. The results are difficult to interpret in terms of a potentiated differentiation of the cells.     

Glutamate and GABA modulate dopamine in the pedunculopontine tegmental nucleus

The pedunculopontine tegmental nucleus (PPTg) has an important anatomical position connecting basal ganglia and limbic systems with motor execution structures in the pons and spinal cord. It receives glutamatergic and GABAergic input and has additional reciprocal connections with mesencephalic dopaminergic neurons, suggesting that the PPTg plays a key role in frontostriatal information processing. In vivo microdialysis in freely moving rats, in combination with behavioral analysis, was used in this study to investigate whether the dopaminergic input can be modulated at the level of the PPTg via N-methyl-d-aspartate (NMDA), α-amino-3-hydroxy-5-methyl-isoxazole-4-propionic acid (AMPA) or GABA_B receptors. Stimulation of the GABA_B receptor decreased dopamine release in the PPTg while that of the AMPA and NMDA receptors increased it. A time-related comparison of the effects of NMDA (0.75 and 1 mM) and AMPA (50 and 25 μM) revealed a more long-lasting effect after AMPA stimulation than after NMDA. However, only the infusion of the GABA_B receptor agonist baclofen (100 and 200 μM) stimulated stereotyped behavior (e.g. sniffing, digging or head movements) and contralateral circling. This study clearly demonstrates that GABAergic as well as glutamatergic terminals in the PPTg are critically involved in the modulation of the dopamine system. Moreover, a decrease in PPTg dopamine via GABA_B receptor stimulation seems to be behaviorally relevant. Electronic Publication     

Spontaneous activity of isolated dopaminergic periglomerular cells of the main olfactory bulb

We examined the electrophysiological properties of a population of identified dopaminergic periglomerular cells of the main olfactory bulb using transgenic mice in which catecholaminergic neurons expressed human placental alkaline phosphatase (PLAP) on the outer surface of the plasma membrane. After acute dissociation, living dopaminergic periglomerular cells were identified by a fluorescently labeled monoclonal antibody to PLAP. In current-clamp mode, dopaminergic periglomerular cells spontaneously generated action potentials in a rhythmic fashion with an average frequency of 8 Hz. The hyperpolarization-activated cation current (Ih) did not seem important for pacemaking because blocking the current with ZD 7288 or Cs+ had little effect on spontaneous firing. To investigate what ionic currents do drive pacemaking, we performed action-potential-clamp experiments using records of pacemaking as voltage command in voltage-clamp experiments. We found that substantial TTX-sensitive Na+ current flows during the interspike depolarization. In addition, substantial Ca2+ current flowed during the interspike interval, and blocking Ca2+ current hyperpolarized the neurons and stopped spontaneous firing. These results show that dopaminergic periglomerular cells have intrinsic pacemaking activity, supporting the possibility that they can maintain a tonic release of dopamine to modulate the sensitivity of the olfactory system during odor detection. Calcium entry into these neurons provides electrical drive for pacemaking as well as triggering transmitter release.     

GABA release induced by aspartate-mediated activation of NMDA receptors is modulated by dopamine in a selective subpopulation of amacrine cells

Glutamate and GABA are the major excitatory and inhibitory neurotransmitters in the CNS, including the retina. In the chick retina, GABA is located in horizontal and amacrine cells and in some cells in the ganglion cell layer. It has been shown that glutamate and its agonists, NMDA, kainate, and aspartate, promote the release of GABA from isolated retina and from cultured retinal cells. Dopamine, the major catecholamine in the retina, inhibits the induction of GABA release by NMDA. Two to seven-day-old intact chicken retinas were stimulated with different glutamatergic agonists and the GABA remaining in the tissue was detected by immunohistochemical procedures. The exposure of retinas to 100 M NMDA for 30 minutes resulted in 50% reduction in the number of GABA-immunoreactive amacrine cells. Aspartate (100 M) treatment also resulted in 60% decrease in the number of GABA-immunoreactive amacrine cells. The number of GABA-immunoreactive horizontal cells was not affected by either NMDA or aspartate. In addition, dopamine reversed by 50% the reduction of the number of GABA-immunoreactive amacrine cells exposed to NMDA or aspartate. Kainate stimulation promoted a 50% reduction in the number of both GABA-immunoreactive amacrine and horizontal cells. Dopamine did not interfere with the kainate effect. While in control and in non-stimulated retinas a continuous and homogeneous immunolabeling was observed throughout the inner plexiform layer, retinas exposed to NMDA, kainate and aspartate displayed only a faint punctate labeling in the inner plexiform layer. It is concluded that, under our experimental conditions, both NMDA and aspartate induce the release of GABA exclusively from amacrine cells, and that the release is modulated by dopamine. On the other hand, kainate stimulates GABA release from both amacrine and horizontal cells with no interference of dopamine.     

A-type K+ current of dopamine and GABA neurons in the ventral tegmental area

A-type K(+) current (I(A)) is a rapidly inactivating voltage-dependent potassium current which can regulate the frequency of action potential (AP) generation. Increased firing frequency of ventral tegmental area (VTA) neurons is associated with the reinforcing effects of some drugs of abuse like nicotine and ethanol. In the present study, we classified dopamine (DA) and GABA VTA neurons, and investigated I(A) properties and the physiological role of I(A) in these neurons using conventional whole cell current- and voltage-clamp recording. DA VTA neurons had a mean firing frequency of 3.5 Hz with a long AP duration. GABA VTA neurons had a mean firing frequency of 16.7 Hz with a short AP duration. For I(A) properties, the voltage-dependence of steady-state I(A) activation and inactivation was similar in DA and GABA VTA neurons. I(A) inactivation was significantly faster and became faster at positive voltages in GABA neurons than DA neurons. Recovery from inactivation was significantly faster in DA neurons than GABA neurons. I(A) current density at full recovery was significantly larger in DA neurons than GABA neurons. In DA and GABA VTA neurons, latency to the first AP after the recovery from membrane hyperpolarization (repolarization latency) was measured. Longer repolarization latency was accompanied by larger I(A) current density in DA VTA neurons, compared with GABA VTA neurons. We suggest that I(A) contributes more to the regulation of AP generation in DA VTA neurons than in GABA VTA neurons.     

Specificity of GABA as a stimulator of dopamine release in rat substantia nigra

The effects of amino acids and the benzodiazepine, flurazepam, on spontaneous and potassium-stimulated [3H]dopamine (DA) release were studied in slices of rat substantia nigra. None of the amino acids tested liberated [3H]DA directly. Flurazepam increased [3H]DA output by 51% and this effect was partially blocked by bicuculline but not by picrotoxin. GABA, L-glutamate and flurazepam, but none of the other compounds, potentiated the release of [3H]DA evoked by 20 mM KCl. The facilitation produced by flurazepam, but not by GABA or L-glutamate, was weakly antagonized by bicuculline and not picrotoxin. GABA did not modify the efflux of [3H]DA induced by low Na+, veratridine, ouabain, calcium ionophore A 23187, amphetamine or exogenous DA.     

Two types of periglomerular cells in the olfactory bulb of the macaque monkey (Macaca fascicularis)

The olfactory bulb (OB) of mammals is the brain region that receives the sensory information coming from the olfactory epithelium. The entrance of the olfactory information occurs in spherical structures of neuropil named olfactory glomeruli and is modulated by a population of interneurons known as periglomerular cells (PG). It has been demonstrated that there are two types of PG in the OB of some macrosmatic mammals, including rats and mice. Type 1 PG (PG-1) receive synapses from the olfactory nerve, whereas type 2 PG (PG-2) do not receive synapses from the olfactory axons. To date, the presence of the two types of PG has not been investigated in microsmatic mammals. In this context, we analyze the presence of PG-1 and PG-2 in the OB of the long-tailed macaque (Macaca fascicularis). For that, we used the enzyme tyrosine hydroxylase, the neuronal isoform of the enzyme nitric oxide synthase and the calcium-binding proteins calbindin D-28k and calretinin as neurochemical markers. Our results demonstrate that the OB of the macaque contains PG-1 and PG-2. A subpopulation of PG-1 expresses tyrosine hydroxylase and another expresses the neuronal isoform of nitric oxide synthase. In addition, a subpopulation of PG-2 expresses calbindin D-28k and another expresses calretinin. Double immunofluorescence demonstrates that there is no colocalization of two markers in the same PG. These results mimic those found in macrosmatic animals. The presence of two types of PG in the glomerular circuits seems to be a key principle for the organization of the OB of mammals.     

Immunocytochemical localization of GABA neurons and dopamine neurons in the rat main and accessory olfactory bulbs

Immunocytochemical localization of GABA neurons and dopamine neurons in the rat olfactory bulb was obtained with sheep antiserum to glutamate decarboxylase (GAD) and rabbit antiserum to tyrosine hydroxylase (TH). GAD-positive neurons include periglomerular cells, granule cells, superficial and deep short axon cells. TH-positive neurons represent periglomerular cells. Two-color immunocytochemistry shows that GABA and dopamine periglomerular cells are separate populations. The accessory olfactory bulb has rare dopamine cells and few superficial short axon cells. Radial gradients of GAD-immunostaining are evident in the main but not in the accessory olfactory bulb.     

Histochemical support for a dopaminergic mechanism in the dendrites of certain periglomerular cells in the rat olfactory bulb

With the Vibratome modification of the Falck-Hillarp technique, and with the indirect immunofluorescence technique for visualizing the first three enzymes in catecholamine synthesis, evidence has been obtained that in the rat olfactory bulb several periglomerular cells are dopaminergic. Both tyrosine hydroxylase and dopadecarboxylase, converting tyrosine to DOPA and DOPA to dopamine (DA), respectively, but not dopamine-β-hydroxylase, converting DA to noradrenaline, are present in periglomerular cell bodies, as well as in their intraglomerular dendrites. These findings suggest that DA may be active at dendrodentritic synapses known to be present between periglomerular and mitral and/or tufted cells.     

Co-transmission in the rat vas deferens: postjunctional synergism of noradrenaline and adenosine 5'-triphosphate

In the isolated prostatic half of the rat vas deferens, joint application of noradrenaline (NA) and adenosine 5'-triphosphate (ATP) produced a contractile response whose magnitude was greatly larger than the addition of the tension generated by the application of each agent alone. The effect of ATP was mimicked by two non-hydrolyzable ATP analogs, but not by GTP, AMP or adenosine. In sympathectomized rats, ATP potentiated NA effects, increasing both the peak tension and the duration of the vas deferens contractile response. The synergism was concentration related. Prazosin antagonized the NA synergism but not the ATP response. Likewise, desensitization of the P2-purinoceptor blocked the ATP synergism without modifying the NA-induced contraction.     

帕金森病大鼠模型纹状体中谷氨酸、γ-氨基丁酸与多巴胺之间的关系

目的:了解帕金森病大鼠模型纹状体内谷氨酸(glutamate,Glu)、γ-氨基丁酸(gama-aminobutyric acid,GABA)和多巴胺(dopamine,DA)之间的关系,从而进一步探讨帕金森病的发病机制。方法:动物分为溶剂对照组、假手术组和帕金森模型组。大脑右侧黑质致密部和前脑内侧束两点注射6-羟基多巴胺(6-hydroxydopamine,6-OHDA)建立帕金森病大鼠模型,溶剂对照组注入生理盐水,假手术组不注射任何药物,采用脑微透析术于建模后第3,4,5,6周连续动态透析大鼠毁损侧纹状体,结合高效液相色谱(HPLC)动态监测各组谷氨酸、GABA和多巴胺的变化。结果:(1)PD组纹状体内多巴胺含量到第5周仅为溶剂对照组和假手术组的1/5;(2)谷氨酸含量随建模时间逐渐升高,到第6周PD组是溶剂对照组和假手术组的1倍以上;(3)GABA含量呈下降趋势,到第6周约降至溶剂对照组、假手术组的1/2。结论:帕金森病大鼠模型纹状体内谷氨酸的变化与多巴胺分泌可能存在某种联系;GABA含量随建模时间的增加而下降。     

Neuropeptide co-release with GABA may explain functional non-monotonic uncertainty responses in dopamine neurons

Co-release of the inhibitory neurotransmitter GABA and the neuropeptide substance-P (SP) from single axons is a conspicuous feature of the basal ganglia, yet its computational role, if any, has not been resolved. In a new learning model, co-release of GABA and SP from axons of striatal projection neurons emerges as a highly efficient way to compute the uncertainty responses that are exhibited by dopamine (DA) neurons when animals adapt to probabilistic contingencies between rewards and the stimuli that predict their delivery. Such uncertainty-related dopamine release appears to be an adaptive phenotype, because it promotes behavioral switching at opportune times. Understanding the computational linkages between SP and DA in the basal ganglia is important, because Huntington’s disease is characterized by massive SP depletion, whereas Parkinson’s disease is characterized by massive DA depletion.     

Neuropharmacology of theophylline induced stuttering: the role of dopamine, adenosine and GABA

Developmental stuttering is a poorly understood speech disorder that starts out in childhood and some individuals continue to stutter throughout their lives. Stuttering is a disruption in smooth and fluent speech. Some stuttering primarily involves vocal blocks, which are spasms of the laryngeal musculature while prolongations, and repetitions of sound occur in other cases. Acquired stuttering, on the other hand, can occur at all ages and can be caused by brain injury and by pharmacological agents. Theophylline-induced stuttering is form of acquired stuttering. It is a rare side effect of theophylline therapy, but it provides interesting clues to the pharmacological mechanisms involved in stuttering. Theophylline-induced stuttering may involve the disrupt the optimal balance between excitatory and inhibitory neurotransmission throughout the brain by inhibiting GABA receptors. The disruption of the optimal balance between excitatory and inhibitory neurotransmission can also cause dysfunction in white matter fiber tracts such as those that connect the Broca's area to the motor cortex. This leads to a hyperexitation of the motor cortex which may mimic the motor cortex hyperexitability that exists in developmental stuttering. Theophylline also enhances dopaminergic neurotransmission through the inhibition of adenosine receptors and this may mimic the hyperdopaminergic state that exists in the brain of developmental stutterers. Theophylline causes the greatest release of dopamine in the basal ganglia through the inhibition of adenosine and GABA receptors. This may also cause dysfunction in the basal ganglia similar in some ways to the dysfunction that exits in developmental stuttering. Pharmacological enhancement of dopaminergic neurotransmission by other drugs been reported to cause stuttering in fluent individuals and to aggrevate dysfluency in stutterers.     

Synaptic organization of the glomerulus in the main olfactory bulb: Compartments of the glomerulus and heterogeneity of the periglomerular cells

According to the combinatorial receptor and glomerular codes for odors, the fine tuning of the output level from each glomerulus is assumed to be important for information processing in the olfactory system, which may be regulated by numerous elements, such as olfactory nerves (ONs), periglomerular (PG) cells, centrifugal nerves and even various interneurons, such as granule cells, making synapses outside the glomeruli. Recently, structural and physiological analyses at the cellular level started to reveal that the neuronal organization of the olfactory bulb may be more complex than previously thought. In the present paper, we describe the following six points of the structural organization of the glomerulus, revealed by confocal laser scanning microscopy and electron microscopy analyses of rats, mice and other mammals: (i) the chemical heterogeneity of PG cells; (ii) compartmental organization of the glomerulus, with each glomerulus consisting of two compartments, the ON zone and the non-ON zone; (iii) the heterogeneity of PG cells in terms of their structural and synaptic features, whereby type 1 PG cells send their intraglomerular dendrites into both the ON and non-ON zones and type 2 PG cells send their intraglomerular dendrites only into the non-ON zone, thus receiving either few synapses from the ON terminals, if present, or none at all; (iv) the spatial relationship of mitral/tufted cell dendritic processes with ON terminals and PG cell dendrites; (v) complex neuronal interactions via chemical synapses and gap junctions in the glomerulus; and (vi) comparative aspects of the organization of the main olfactory bulb.     

GABA and GABA receptors in invertebrates

GABA is the major inhibitory transmitter at invertebrate synapses in both the central and peripheral nervous systems. The receptors for GABA are well characterised electrophysiologically in a wide variety of invertebrate organisms but their biochemical and pharmacological profiles are less well defined. In general invertebrate GABA receptors are less sensitive to bicuculline than are vertebrate GABA_A receptors. There is much evidence that invertebrate GABA_A receptors have benzodiazepine modulatory sites but their pharmacological profile is distinct from that of vertebrate GABA_A receptors. The insect neuronal GABA_A receptor has been identified as the site of action of the cyclodiene insecticides and the avermectins may also act on these receptors. Molecular cloning experiments now in progress will reveal the detailed relationships between the invertebrate receptors and their mammalian counterparts.     

A simple model of the interactions of dopamine, acetylcholine and GABA in movement disorders seen in psychiatry

Many of the movement disorders seen in psychiatric patients are determined, in large part, by the dynamic balance of dopamine and acetylcholine in the basal ganglia. Gamma aminobutyric acid has effects on various neurotransmitters in the brain including dopamine and acetylcholine and is therefore relevant to a discussion of movement disorders. Laboratory and clinical data on the effects of gamma aminobutyric acid on dopamine and acetylcholine are discussed, and a simple model of the interactions of these transmitters, with respect to movement disorders seen in psychiatry, is discussed. Some clinical evidence that is consistent with this model is discussed. Guidelines for the use of gamma aminobutyric acid--like drugs for movement disorders seen in clinical practice are also discussed.     

State-dependent changes in glutamate, glycine, GABA, and dopamine levels in cat lumbar spinal cord

Recent studies have indicated that the glycine receptor antagonist strychnine and the gamma-aminobutyric acid type A (GABA A) receptor antagonist bicuculline reduced the rapid-eye-movement (REM) sleep-specific inhibition of sensory inflow via the dorsal spinocerebellar tract (DSCT). These findings imply that the spinal release of glycine and GABA may be due directly to the REM sleep-specific activation of reticulospinal neurons and/or glutamate-activated last-order spinal interneurons. This study used in vivo microdialysis and high-performance liquid chromatography analysis techniques to provide evidence for these possibilities. Microdialysis probes were stereotaxically positioned in the L3 spinal cord gray matter corresponding to sites where maximal cerebellar-evoked field potentials or individual DSCT and nearby spinoreticular tract (SRT) neurons could be recorded. Glutamate, glycine, and GABA levels significantly increased during REM sleep by approximately 48, 48, and 14%, respectively, compared with the control state of wakefulness. In contrast, dopamine levels significantly decreased by about 28% during REM sleep compared with wakefulness. During the state of wakefulness, electrical stimulation of the nucleus reticularis gigantocellularis (NRGc) at intensities sufficient to inhibit DSCT neuron activity, also significantly increased glutamate and glycine levels by about 69 and 45%, respectively, but not GABA or dopamine levels. We suggest that the reciprocal changes in the release of glutamate, glycine, and GABA versus dopamine during REM sleep contribute to the reduction of sensory inflow to higher brain centers via the DSCT and nearby SRT during this behavioral state. The neural pathways involved in this process likely include reticulo- and diencephalospinal and spinal interneurons.     

Tyrosine hydroxylase and 28K-vitamin D-dependent calcium binding protein are localized in different subpopulations of periglomerular cells of the rat olfactory bulb

Successive incubations of rat olfactory bulb cryostat sections with antibodies against the chick 28K-vitamin D-dependent calcium (Ca) binding protein and tyrosine hydroxylase were performed, and the distribution of the label was examined in the fluorescence microscope. Both antibodies labeled cells and processes in highest number in the glomerular layer. The two immunopositivities were not co-localized in the same neurons. This suggests that the presence of this Ca-binding protein is not a necessary prerequisite for dopaminergic neurotransmission in periglomerular neurons.