Release Of Endogenous Dopamine Research Articles

Regulation of endogenous dopamine release in amphibian retina by melatonin: the role of GABA.

In the retina of the African clawed frog (Xenopus laevis), endogenous dopamine release increases in light and decreases in darkness. Exogenous melatonin and several chemical analogs of melatonin suppressed light-evoked dopamine release from frog retina in a concentration-dependent manner. The rank order of potency for inhibition of light-evoked dopamine release was melatonin >> 5-methoxytryptamine > or = N-acetylserotonin > 5-methoxytryptophol >>> serotonin. Melatonin did not suppress dopamine release below levels seen in darkness. The putative melatonin receptor antagonist luzindole inhibited the effect of melatonin. Luzindole enhanced dopamine release in darkness but had little effect in light. These data suggest a role for endogenous melatonin in dark-induced suppression of retinal dopamine. Picrotoxin and bicuculline, GABA-A receptor antagonists, blocked melatonin-induced suppression of dopamine release. In the presence of melatonin, bicuculline was significantly less potent in stimulating dopamine release. These results suggest that melatonin enhances GABAergic inhibition of light-evoked dopamine release. This mechanism may underlie the light/dark difference in dopamine release in vertebrate retina.

Effects of domperidone on neonatal and adult carotid chemoreceptors in the cat.

It has been postulated that the weak carotid chemoreceptor responses of neonatal mammals may be due to inhibition produced by high levels of endogenous dopamine release or exaggerated sensitivity to dopaminergic inhibition. This was studied by measuring the effect of domperidone, a selective dopamine D2-receptor antagonist, on the carotid chemoreceptor response to O2 and CO2 in anesthetized neonatal and adult cats. The animals were exposed to four levels of isocapnic O2 (arterial PO2 of approximately 35-45, 55-65, 80-90, > 300 Torr) and four levels of isoxic CO2 (end-tidal PCO2 of approximately 21, 40, 58, and 78 Torr) before and after D2-receptor blockade. Whole nerve activity was recorded from the carotid sinus nerve (CSN). Both neonatal and adult cats increase CSN activity during hypoxia and hypercapnia (P < 0.001). Domperidone caused an increase in CSN activity at all O2 levels in adults (P < 0.01) but only during hypoxia in neonates (P < 0.001). Domperidone caused an increase in CSN activity during normo- and hypercapnia in adults but only during hypercapnia in neonates (P < 0.001). Domperidone approximately doubled an index of hypoxic sensitivity in the normoxia-hypoxia range (100 to 40 Torr) in the neonatal group but had little effect on sensitivity to hypoxia in adults. We conclude that the inhibitory role of endogenous dopamine in the carotid chemoreceptors changes with postnatal development.

Regulation of endogenous dopamine release in amphibian retina by gamma-aminobutyric acid and glycine.

Endogenous dopamine release in the retina of the African clawed frog (Xenopus laevis) increases in light and decreases in darkness. The roles of the inhibitory amino acid transmitters gamma-aminobutyric acid (GABA) and glycine in regulating this light/dark difference in dopamine release were explored in the present study. Exogenous GABA, the GABA-A receptor agonist muscimol, the GABA-B receptor agonist baclofen, and the GABA-C receptor agonist cis-aminocrotonic acid (CACA) suppressed light-evoked dopamine overflow from eyecups. The effects of GABA-A and -B receptor agonists were selectively reversed by their respective receptor-specific antagonists, whereas the effect of CACA was reversed by the competitive GABA-A receptor antagonist bicuculline. The benzodiazepine diazepam enhanced the effect of muscimol on light-evoked dopamine release. Both GABA-A and -B receptor antagonists stimulated dopamine release in light or darkness. Bicuculline was more potent in light than in darkness. These data suggest that retinal dopaminergic neurons are inhibited by GABA-A and -B receptor activation in both light and darkness but that GABA-mediated inhibitory tone may be greater in darkness than in light. Exogenous glycine inhibited light-stimulated dopamine release in a concentration-dependent and strychnine-sensitive manner. However, strychnine alone did not increase dopamine release in light or darkness, nor did it augment bicuculline-stimulated release in darkness. Additionally, both strychnine and 7-chlorokynurenate, an antagonist of the strychnine-insensitive glycine-binding site of the N-methyl-D-aspartate subtype of glutamate receptor, suppressed light-evoked dopamine release. Thus, the role of endogenous glycine in the regulation of dopamine release remains unclear.

PEC-60 increases dopamine but not GABA release in the dorsolateral neostriatum of the halothane anaesthetized rat. An in vivo microdialysis study

The effect of striatal perfusion with the intestinal peptide PEC-60 on endogenous dopamine (DA) and γ-aminobutyric acid (GABA) release in the dorsolateral striatum and GABA release in the globus pallidus was monitored using in vivo microdialysis in the halothane anaesthetized rat. The results show that PEC-60 (100 nM) increases DA release in the dorsolateral striatum without influencing GABA release in the dorsolateral striatum or in the globus pallidus. In addition, PEC-60 failed to influence the extracellular striatal 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) levels. The PEC-60 induced increase in striatal DA was abolished by the addition of tetrodotoxin (1 μM) to the perfusion medium. These data suggest that PEC-60 plays a role in modulating striatal DA release but not DA metabolism and that this effect is primarily targeted on the presynaptic DA terminals of the nigrostriatal DA pathway rather than on the postsynaptic striatopallidal GABA projection neurons in the dorsolateral striatum.

Direct monitoring of dopamine and 5-HT release in substantia nigra and ventral tegmental area in vitro.

Fast-scan cyclic voltammetry with carbon fibre microelectrodes was used to detect endogenous dopamine (DA) and 5-hydroxytryptamine (5-HT) release from three distinct regions of guinea-pig mid-brain in vitro: rostral and caudal substantia nigra (SN) and the ventral tegmental area (VTA). Previous electrophysiological studies have demonstrated that cells of the caudal SN and the VTA have similar characteristics, whereas cells in the rostral SN have distinctly different properties. In the present study, we confirmed that each region has tyrosine hydroxylase-positive neurons and determined, using high-performance liquid chromatography, that DA levels were similar in rostral and caudal SN, but lower in SN than in VTA. In each region, application of veratrine, which was shown by intracellular recordings to have a reversible depolarising action, evoked a signal attributable to DA and distinguishable from that of 5-HT. Release signals were monitored every 250 ms with a spatial resolution of less than 50 microns.l DA release was calcium-dependent and was not detectable in a catecholamine-poor area such as the cerebellum, or in mid-brain tissue pre-treated with reserpine. Within the normal mid-brain, the amount of DA released was correlated with tissue content in that it was higher in the VTA than in either region of SN. It is concluded that DA released from somato-dendritic parts of mid-brain neurons exhibits site-specific variation. This is the first report of direct monitoring of DA and 5-HT release from these regions with in situ electrodes and demonstrates the utility of fast-scan cyclic voltammetry to investigate the mechanisms and possible non-classical functions of somato-dendritic DA release.

The weaver mutant mouse as a model of nigrostriatal dysfunction.

The weaver mutant mouse has a genetic defect that results in the loss of dopamine neurons in the nigrostriatal pathway. Striatal tyrosine hydroxylase and dopamine content are reduced by 60-70%, and dopamine uptake is reduced by as much as 95%. Deficits in all three of these striatal dopamine markers are seen as early as postnatal d 3. The striatal dopamine systems in the weaver apparently have the ability to compensate for this dopamine deficit. Thus, in the weaver, in vitro resting release, as well as amphetamine-evoked fractional release of endogenous dopamine are increased. An additional change seen in the weaver striatum is an elevated serotonin content. These alterations may play an adaptive role in attempting to compensate for the dopamine loss. In summary, the weaver mutant mouse has dramatic deficits in the nigrostriatal pathway, but also seems to develop certain adaptive mechanisms in dopaminergic and other transmitter systems that may compensate functionally for the dopamine deficit. Thus, the weaver mouse provides a unique animal model for studying naturally induced neuronal degeneration that complements those models using surgical and pharmacological protocols.

Effect of dopamine on hypoxic ventilatory response of sedated piglets with intact and denervated carotid bodies

To determine whether the neonatal hypoxic ventilatory depression is in part produced by an increased endogenous dopamine release that can depress the activity of central and peripheral chemoreceptors, 31 sedated and spontaneously breathing newborn piglets [age 5 +/- 1 (SD) days; weight 1.7 +/- 0.4 kg] were randomly assigned to an intact carotid body or a chemodenervated group. Minute ventilation (VE), arterial blood pressure, and cardiac output (CO) were measured in room air before infusion of saline or the dopamine antagonist flupentixol (0.2 mg/kg i.v.) and 15 min after drug infusion and were repeated after 10 min of hypoxia (inspiratory O2 fraction = 0.10). VE increased significantly after 10 min of hypoxia in the piglets that received flupentixol independent of whether the carotid bodies were intact or denervated. However, the increase in VE was largest and sustained throughout the 10 min of hypoxia only in the intact carotid body flupentixol group. As expected, the initial increase in VE with hypoxia was abolished by carotid body denervation. Changes in arterial blood gases, CO, and mean arterial blood pressure with hypoxia were not different among groups. These results demonstrate that flupentixol reverses the late hypoxic decrease in VE, acting through peripheral and central dopamine receptors. This effect is not related to changes in cardiovascular function or acid-base status.

Subchronic treatment with the neuroleptic-like peptide desenkephalin- γ-endorphin may decrease dopaminergic neurotransmission in the nucleus accumbens of rats

In rats, subchronic administration of desenkephalin-γ-endorphin (DEγE) into the nucleus accumbens or subcutaneously for 10 days resulted in hypoactivity. Intra-accumbens administration caused a significant reduction in the nucleus accumbens tissue levels of the dopamine (DA) metabolites 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA). Systemic administration of DEγE decreased DOPAC and 5-hydroxyindoleacetic acid (5-HIAA) levels in nucleus accumbens tissue. Subchronic subcutaneous DEγE treatment reduced the basal release of [ 3H]DA from rat nucleus accumbens slices in vitro and the basal release of endogenous DA and DOPAC in vivo as assessed with on-line dialysis in the nucleus accumbens of freely moving rats. The DA agonist N, N-dipropyl-7-hydroxy-2-aminotetralin (DP-7-ATN) was equally effective in inhibiting [ 3H]DA release elicited by electrical stimulation from slices of subchronically DEγE and placebo treated rats. Administration of a small dose of apomorphine caused similar reductions of the in vivo release of DA and DOPAC in both placebo and DEγE treated rats. These results indicate that subchronic DEγE treatment may decrease dopaminergic neurotransmission in the nucleus accumbens. This effect is probably not due to alterations in the sensitivity of presynaptically located DA autoreceptors mediating DA release in vitro and in vivo.

Early abnormalities of retinal dopamine pathways in rats with hereditary retinal dystrophy.

The dopaminergic pathway that affects rod-driven horizontal cells has been studied in the Royal College of Surgeons (RCS) rat during the period preceding photoreceptor degeneration (postnatal day 17-24). The experiments were performed by intracellular recording from single horizontal cells in vitro. Horizontal cells from the recessive control animals (postnatal day 17-24) were depolarized by dopamine (10 microM) and hyperpolarized by the D1 antagonist SCH 23,390 (10 microM). In contrast, cells from age-matched dystrophic retinas, though depolarized by dopamine, were unaffected by SCH 23390 (10-100 microM), suggesting a significant reduction in the level of endogenous dopamine release. Histologic examination for catecholaminergic neurons revealed no differences in either the cell number or anatomy between the retinas of the control and dystrophic animals. Furthermore, perfusion of the control retinas with melatonin (500 nM-1 microM) yielded response characteristic of the dystrophic type. In the period preceding degeneration, the RCS retina thus displays a discrete abnormality in dopaminergic pathways, such that there is a gross reduction in endogenous dopamine release below that required to activate D1 receptors. Since melatonin levels have been shown to be high in these retinas, we propose that abnormalities in the dopamine-melatonin systems give rise to an electrophysiologic deficit in the postphotoreceptoral retina of the RCS rat.

Nitric oxide modulates endogenous dopamine release in bovine retina.

A study of the possible modulation by nitric oxide (NO) of endogenous dopamine (DA) release was performed in bovine retina in vitro. NO synthase activity was measured in retinal homogenates and totally blocked by L-nitroarginine methyl ester (L-NA). Intact retinas were also exposed to hydroxylamine, an NO-generator which significantly decreased both basal and potassium-induced liberation of DA. In contrast, dibutyryl cGMP was ineffective. Furthermore, L-NA was able to increase basal DA release, its effects being potentiated in calcium free medium. Taken together, these results suggest that endogenous NO regulates DA release via cGMP independent mechanisms.

P-162 - Electrically stimulated release of endogenous dopamine and amino acids from rat brain slices under ischemia-like condition.

P-162 - Electrically stimulated release of endogenous dopamine and amino acids from rat brain slices under ischemia-like condition.

Investigation of the ontogenetic patterns of rat hypothalamic dopaminergic neurone morphology and function in vitro.

Using fetal rat hypothalamic cells in primary culture maintained in a serum-free defined medium we have investigated the morphological and functional development of the dopamine (DA)-containing neurones intrinsic to the hypothalamus. Immunocytochemical studies demonstrated the presence of three morphologically distinct subtypes of tyrosine hydroxylase-immunopositive neurones. On day 3 in vitro unipolar, bipolar and multipolar cell types were apparent. The latter two subtypes persisted to later days in culture and increased both in perikarya size and neurite length. All subtypes have been shown to have correlates in vivo. Biochemical studies employing [3H]DA demonstrated a time- and temperature-dependent uptake mechanism within the cultures which was significantly attenuated by the uptake inhibitors benztropine and nomifensine in a dose-dependent manner. [3H]DA was also released under both basal and 56 mmol K+/l-stimulated conditions and the magnitude of the response was reduced by exclusion of calcium from the release medium. The amount of [3H]DA accumulated and released by the cultural cells increased with the age of the culture, suggesting functional maturation of the DA-containing neurones within this preparation. The role of oestradiol-17 beta in regulating hypothalamic dopaminergic function was also investigated both indirectly with the use of [3H]DA and by direct measurement of endogenously synthesized DA using high-performance liquid chromatography coupled with electrochemical detection. Both uptake and release of [3H] and release of endogenous DA were significantly modulated by the concentration of steroid in the defined medium. These results demonstrate that hypothalamic dopaminergic neurones, when maintained in primary culture, undergo morphological and functional maturation which have several correlates in vivo. In addition, we have demonstrated that at least one sub-population of dopaminergic neurones within this preparation is responsive to oestradiol-17 beta. As DA is considered to be a vital component in the regulation of neuroendocrine activity we suggest that this model is valuable for the investigation of the functional development of the DA systems of the hypothalamus and the relationship existing between neurotransmitters, neuropeptides and neuroactive steroids.

Modulation of endogenous dopamine release in the turtle retina: effects of light, calcium, and neurotransmitters.

In the turtle retina, dopamine has been observed in a small population of amacrine cells. Whereas the effect of dopamine has been intensively studied, knowledge about the release of this transmitter and the neuronal control of its release are still poorly understood. We therefore decided to study the release of endogenous dopamine. Isolated retinas were superfused with Ringer's solutions and stimulated with increased potassium, light, or drugs which interfere with neurotransmitter systems. Dopamine was analyzed by using aluminum-oxide extraction and high-pressure liquid chromatography (HPLC) with electrochemical detection. Increased potassium (25 mM) caused a five-fold increase in the basal release. When calcium was replaced by cobalt, no increase was induced by 25 mM potassium. Flickering light increased the basal release of endogenous dopamine by a factor of three. The effect of flickering light was greater in the presence of additional steady background illumination. Kainate (10 microM), an agonist for excitatory amino acids, doubled the basal dopamine release. Bicuculline (10 microM), a gamma-amino butyric acid (GABA) antagonist, increased the release to about six times the basal level. Naloxone (10 microM), an opiate antagonist, increased the release to eight times the basal level. These findings suggest that dopamine is released from amacrine cells in the turtle retina in a calcium-dependent manner, which is most likely a vesicular release. Dopamine release is induced by flickering light vs. darkness and vs. steady background illumination. A moderate background illumination alone does not significantly increase basal dopamine release.(ABSTRACT TRUNCATED AT 250 WORDS)

The effects of selective activation of the 5-HT 3 receptor with m-chlorophenylbiguanide on sleep and wakefulness in the rat

The effects of the 5-HT 3 receptor agonist, m-chlorophenylbiguanide, were compared with those of the 5-HT 3 receptor antagonist, MDL 72222, in rats implanted with electrodes for chronic sleep recordings. m-Chlorophenylbiguanide (12.5–50.0 μg) injected into the left lateral ventricle increased wakefulness and rapid eye movement (REM) sleep latency, whereas slow wave sleep, REM sleep and the number of REM periods were reduced. MDL 72222 (0.1–1.0 mg/kg, s.c.) induced a delayed and dose-dependent increase of slow wave sleep. Pretreatment with MDL 72222 (0.1–0.5 mg/kg) prevented the effects of m-chlorophenylbiguanide (50 μg) on wakefulness and sleep. It is suggested that the increase of wakefulness after 5-HT 3 receptor activation could be related to the release of endogenous serotonin and dopamine.

Glutamate stimulates dopamine release from cortical and limbic rat brain in vitro

In vitro superfusion studies were performed to compare the glutamatergic control of dopamine release from minislices of rat striatum, olfactory tubercle and frontal cortex. In the absence of Mg 2+, release of endogenous dopamine from olfactory tubercle and striatum, and [ 3H]dopamine from frontal cortex, was stimulated by glutamate (at concentrations of 0.3, 3 and 1 mM respectively). In all three areas the response to N-methyl-D-aspartate (NMDA) was greater than that to kainate, implying mediation largely by NMDA receptors, with non-NMDA receptors contributing to a smaller extent. This was confirmed by antagonist studies in the frontal cortex, where the response to glutamate was largely (70%) blocked by 2-amino-5-phosphonopentanoic acid (AP5), and further blocked by AP5 + 6,7-dinitroquinoxaline-2,3-dione (DNQX). In the olfactory tubercle and striatum, glutamate was as effective as NMDA, but in the frontal cortex the response to NMDA was significantly larger than that to glutamate.

Chromogranin A inhibits retinal dopamine release

Chromogranin A (CGA), a peripherally active prohormone, is a soluble component in the secretory granules of many endocrine tissues and is cosecreted with their peptide or amine hormones. Using an antibody prepared against purified rat adrenal CGA, immunostaining was localized to the inner and outer plexiform layers of the rat retina and to selected ganglion cells. Exogenous CGA (purified from human adrenals) when applied to perfused rat retina potently inhibited the potassium-induced release of endogenous dopamine (DA). This action was dose-dependent, with an IC 50 of 3 nM; at 100 nM CGA retinal DA release was completely abolished.

Dissociation of the effects of amphetamine and quinpirole on dopamine release in the nucleus accumbens following behavioural sensitization

Behavioural sensitization to the locomotor stimulant effect of (+)-amphetamine or quinpirole was induced in rats by intermittent drug administration. Once established, endogenous dopamine release (DA) was measured in slices containing nucleus accumbens using fast cyclic voltammetry. DA release induced by single pulse electrical stimulation did not differ between vehicle-, (+)-amphetamine-or quinpirole-treated groups. Multiple pulse stimulation resulted in enhanced DA release in quinpirole-sensitized rats and an attenuation of DA release in (+)-amphetamine-sensitized rats. In the presence of sulpiride, DA release was increased, at low stimulation frequencies, in vehicle- and quinpirole-treated animals, but not in amphetamine-treated animals. The sensitivity of axon terminal D2 DA receptors was assessed in vitro by measuring the concentration of quinpirole required to inhibit single pulse release of DA by 50% (EC(50)). Quinpirole EC(50) was significantly increased in the quinpirole-treated animals and significantly attenuated in the (+)-amphetamine-treated animals. The results suggest that the increase in DA release following quinpirole may arise from the desensitization of release-regulating D2 autoreceptors in the nucleus accumbens. The sensitization of axon terminal D2 autoreceptors and the decrease in DA release following behavioural sensitization with (+)-amphetamine is difficult to reconcile with a unitary explanation of behavioural sensitization to both quinpirole and (+)-amphetamine. It is suggested that the effects following (+)-amphetamine may be secondary to decreased axon traffic caused by DA displacement in the ventral tegmental area, and that the drugs examined in this study may induce behavioural sensitization by different mechanisms at different sites.

A compartmentalized chamber for studying dopamine neurons in an hypothalamo-pituitary explant

An in vitro technique was developed which could be used to study whole neurons of the tuberohypophyseal dopaminergic (THDA) tract. Explants containing the medial basal hypothalamus (MBH; THDA cell bodies), pituitary stalk (THDA axons) and posterior pituitary (PP; THDA nerve terminals) were carefully dissected and placed in specially designed chambers. The chambers consist of separate compartments for the incubation of the MBH and PP while maintaining an intact pituitary stalk which traverses through a notched barrier. Dopamine (DA) synthesis in the explants was constant for 3 h and was significantly elevated by electrical stimulation. Electrical stimulation of both the PP and MBH increased endogenous DA release from the PP. DA release in response to potassium depolarization and dopaminergic drugs was significantly lower in explants than in the isolated PP even though spontaneous release was similar. Conclusions: (1) this method is suitable for studying intact THDA neurons in vitro, and (2) intact THDA neurons respond differently to various stimuli than their excised terminals.

Electrically induced release of endogenous noradrenaline and dopamine from brain slices: pseudo-one-pulse stimulation utilized to study presynaptic autoinhibition.

Slices of rat hypothalamus (noradrenaline experiments) or rabbit caudate nucleus (dopamine experiments) were prepared, superfused, and field-stimulated using series of monophasic rectangular pulses. Noradrenaline, dopamine and the main dopamine metabolite, dihydroxyphenylacetic acetic acid (DOPAC), were determined using HPLC with electrochemical detection. Electrical stimulation was performed using the following protocols: 1) 4 pulses delivered at 100 Hz; this type of stimulation is referred to as pseudo-one-pulse stimulation (POP); its short duration of only 32 ms does not allow the development of autoinhibition; 2) 2 bursts of 4 pulses at 100 Hz, delivered 1 s apart (2-POP-stimulation); 3) 8 pulses at 1 Hz (dopamine experiments only); 4) 36 pulses at 3 Hz. Noradrenaline experiments. The alpha 2-adrenoceptor antagonist yohimbine (1 mumol/l) did not enhance noradrenaline overflow following POP stimulation, but enhanced the overflow following 2-POP-stimulation by about 50% and that following 36-pulse-stimulation by almost 100%. Dopamine experiments. The D2-dopamine receptor antagonist sulpiride (3 mumol/l) facilitated the overflow of dopamine elicited with 2-POP-stimulation (66%), 8 pulses/1 Hz (92%), and 36 pulses/3 Hz (140%). It did not significantly facilitate the overflow of dopamine following POP-stimulation (19%). The overflow of DOPAC was not, or only slightly, increased by electrical stimulation, and its spontaneous outflow was more than three times higher than that of dopamine. Furthermore, the electrically induced overflow of dopamine did not exceed the outflow of DOPAC at any of the stimulation conditions employed. The results of the present study bear out important claims of the autoreceptor theory and confirm the data obtained in previous experiments using labelled transmitters.

Neuromodulation of pigment movement in the RPE of normal and 6-OHDA-lesioned goldfish retinas.

The role of dopamine as the endogenous signal-initiating light-dependent changes in the distribution of pigment granules in goldfish retinal pigment epithelium was investigated. In normal retinas, light adaptation resulted in the dispersion of pigment granules. This effect of light was mimicked by the intraocular injection of dopamine or serotonin, which is thought to increase endogenous dopamine release, into dark-adapted eyes. The effect of light, dopamine, or serotonin on dark-adapted retinas was blocked by the dopamine receptor antagonists haloperidol and sulpiride. However, lesioning the endogenous source of retinal dopamine, by prior intraocular injection of 6-hydroxydopamine (6-OHDA), did not block the dispersion of pigment granules in light-adapted retinas. No significant differences in pigment dispersion were noted between unlesioned and lesioned light- or dark-adapted retinas. However, the effect of light on pigment dispersion was no longer blocked by haloperidol or sulpiride in 6-OHDA lesioned animals. Dopamine and serotonin mimicked the effect of light when injected into lesioned dark-adapted eyes, but their effects were also not blocked by haloperidol or sulpiride. These results suggest that dopamine, acting on D2 receptors, is sufficient to induce pigment migration in unlesioned animals. In 6-OHDA-lesioned animals, however, pigment migration is mediated by a receptor mechanism other than D2.