The electrophysiological actions of dopamine and dopaminergic drugs on neurons of the substantia nigra pars compacta and ventral tegmental area.

The dopaminergic neurons of the substantia nigra pars compacta and ventral tegmental area play a crucial role in regulating movement and cognition respectively. Several lines of evidence suggest that a degeneration of dopaminergic cells in the substantia nigra produces the symptoms of Parkinson's disease. On the other hand, a hyperactivity of the dopaminergic transmission in the brain induces dyskinesia, dystonia and psychosis. It is also well established that the euphoric and rewarding responses evoked by drugs of addiction, such as amphetamine and cocaine, are mediated by central dopamine systems. Electrophysiological experiments which study the activity of single dopaminergic neurons in the ventral mesencephalon have shown that dopamine and dopaminergic drugs reduce the firing frequency of these cells. This is due to the stimulation of D2-D3 autoreceptors and to a hyperpolarization of the membrane produced by an increase in potassium conductance. In addition, substances which increase the release (amphetamine), the synthesis (levodopa) or block the uptake (cocaine, nomifensine, amineptine) of dopamine in the brain inhibit the firing activity of the dopaminergic cells throughout dopamine-mediated mechanisms. In this review, we will briefly examine the literature concerning the physiological and behavioural responses caused by dopamine and dopaminergic agents on the dopaminergic neurons of the ventral mesencephalon. Our conclusion suggests that the electrophysiological actions of dopamine and dopamine-related drugs on dopaminergic cells in the ventral mesencephalon might be indicative of the pharmacological effects of these agents on the brain.     

Mutant PINK1 upregulates tyrosine hydroxylase and dopamine levels,leading to vulnerability of dopaminergic neurons

PINK1 mutations cause autosomal recessive forms of Parkinson disease (PD). Previous studies suggest that the neuroprotective function of wild-type (WT) PINK1 is related to mitochondrial homeostasis. PINK1 can also localize to the cytosol; however, the cytosolic function of PINK1 has not been fully elucidated. In this study we demonstrate that the extramitochondrial PINK1 can regulate tyrosine hydroxylase (TH) expression and dopamine (DA) content in dopaminergic neurons in a PINK1 kinase activity-dependent manner. We demonstrate that overexpression of full-length (FL) WT PINK1 can downregulate TH expression and DA content in dopaminergic neurons. In contrast, overexpression of PD-linked G309D, A339T, and E231G PINK1 mutations upregulates TH and DA levels in dopaminergic neurons and increases their vulnerability to oxidative stress. Furthermore transfection of FL WT PINK1 or PINK1 fragments with the PINK1 kinase domain can inhibit TH expression, whereas kinase-dead (KD) FL PINK1 or KD PINK1 fragments upregulate TH level. Our findings highlight a potential novel function of extramitochondrial PINK1 in dopaminergic neurons. Deregulation of these functions of PINK1 may contribute to PINK1 mutation-induced dopaminergic neuron degeneration. However, deleterious effects caused by PINK1 mutations may be alleviated by iron-chelating agents and antioxidant agents with DA quinone-conjugating capacity.     

Effect of dopaminergic drugs on hypothalamic and pituitary immunoreactive beta-endorphin concentrations in the rat

Beta-endorphin concentrations have been evaluated in the hypothalamus, pituitary lobes and plasma after 1-and 3-week treatment with 2-Br-alpha-ergocriptine or lisuride, two potent dopaminergic drugs. Hypothalamic beta-endorphin concentrations were significantly decreased after the administration of the dopaminergic agents for 1 or 3 weeks. Similarly, beta-endorphin concentrations decreased in the neurointermediate lobe and plasma. After gel chromatography, it appeared that in the anterior pituitary, beta-lipotropin concentrations were unchanged or lightly increased concomitantly with a decrease of beta-endorphin. Our data indicate that, both in the hypothalamus and the neurointermediate pituitary lobe, beta-endorphin is under an inhibitory dopaminergic tone. The latter may also play a role in inhibiting beta-endorphin cleavage from beta-lipotropin in the anterior pituitary.     

Mesostriatal lesions and endothelial function: Effects of forced movements and hypoxia on levels of circulating endothelins

We studied rats with a hemi-Parkinsonian model created by lesions of mesostriatal dopaminergic neurons. Correlation between the extent of lesions and intensity of forced rotation movements (apomorphine (APO)-induced rotations, i.e., exercise) was investigated. The levels of endothelins (ET) in the circulation after rotation under normoxic and hypoxic conditions were measured. In normoxia, the rotation sessions were associated with significant decreases in circulating ET levels, from 21.32±4.06 to 9.32±1.08 pg/ml. Although hypoxia had no effect on the frequency of rotations induced by APO, it significantly attenuated a decrease in the ET levels in the circulation of rats with the most extensive lesions. We suggest that the mesostriatal dopaminergic system modulates endothelial function by an active process. This may have relevance for some clinical manifestations of disordered striatal function observed in humans.     

Clinical aspects of dopamine agonists and antagonists.

Dopamine and its specific receptors are widely distributed in man. Body regions where dopaminergic activity is of special pharmacologic interest include the basal ganglions, hypothalamus, chemoreceptor trigger zone, other less well defined areas in the central nervous system, and the renal and cardiovascular systems. The search for dopaminergic agents to modify these systems in disease states has depended heavily on in vitro and in vivo bioassays. These assays involving receptor binding, enzyme activation, smooth muscle and neuronal excitation, and modification of animal behavior have provided physicians with important therapeutic tools. Indeed, the introduction of levodopa for the treatment of Parkinson's disease and of the phenothiazines and related drugs for schizophrenia and psychosis has been a hallmark of neuropharmacologic research. However, the maximal benefits that these drugs may afford have not yet been realized due to an inadequate understanding of disease processes and a relative lack of specificity of drug action.     

Selective vulnerability of dopaminergic neurons to microtubule depolymerization

Parkinson disease (PD) is characterized by the specific degeneration of dopaminergic (DA) neurons in substantia nigra and has been linked to a variety of environmental and genetic factors. Rotenone, an environmental PD toxin, exhibited much greater toxicity to DA neurons in midbrain neuronal cultures than to non-DA neurons. The effect was significantly decreased by the microtubule-stabilizing drug taxol and mimicked by microtubule-depolymerizing agents such as colchicine or nocodazole. Microtubule depolymerization disrupted vesicular transport along microtubules and caused the accumulation of dopamine vesicles in the soma. This led to increased oxidative stress due to oxidation of cytosolic dopamine leaked from vesicles. Inhibition of dopamine metabolism significantly reduced rotenone toxicity. Thus, our results suggest that microtubule depolymerization induced by PD toxins such as rotenone plays a key role in the selective death of dopaminergic neurons.     

Possible Mechanism of Interaction of GABAergic-Adenosinergic Systems in the Regulation of Theophylline-Induced Locomotor Activity Under Its Nontolerant and Tolerant Conditions

The measurement of locomotor activity (LA) of theophylline (Th) nontolerant (10 mg/kg, p.o.) rats using agonist and antagonist of different neurotransmitters either in single or in their different combinations suggest that an inhibition of central GABAergic activity as well as adenosinergic and serotonergic activities through the stimulation of dopaminergic activity followed by an inhibition of cholinergic system may stimulate LA in Th nontolerant condition. Further, it is suggested that the development of tolerance to Th restored the LA to control value may be due to an activation of adenosinergic system which possibly withdrew the inhibition occurred in central cholinergic, GABAergic and serotonergic activities followed by the modulation of dopaminergic system.     

Dopamine and alcohol relapse: D1 and D2 antagonists increase relapse rates in animal studies and in clinical trials

A considerable number of animal studies on the effects of dopaminergic agents on alcohol intake behavior have been performed. Acute alcohol administration in rats induces dopamine release in the caudate nucleus and in the nucleus accumbens, an effect related among others to reinforcement. It has been repeatedly suggested that D1 and D2 receptor activation mediates reward. As alcohol consumption and dopaminergic transmission seem to have a close relationship, all kinds of dopaminergic agents may be regarded as putative therapeutics for preventing relapse. In a prospective European double-blind multicenter clinical trial, comparing the D1, D2, D3 antagonist flupenthixol and placebo in 281 chronic alcohol-dependent patients (27.4% women), the application of the Lesch typology made an outcome differentiation possible. It could be shown in which patients flupenthixol administration was followed by a significantly higher relapse rate and in which patient groups no differences were found when compared to placebo.     

Localization and induction in early development of Xenopus

Our experimental results, as well as those of others, lead us to suggest the following steps in the dorsalization and axialization of the Xenopus egg and embryo: the sperm aster determines the direction of rotation of the cortex relative to the deeper cytoplasm (endoplasm); the rotation of the cortex activates latent dorsalizing-axializing agents in the vegetal hemisphere. The extent of rotation determines the amount of activation. The direction of rotation determines the location of the activated agents. The activated agents determine the level of mesoderm-inducing activity of the vegetal cells cleaved from that cytoplasmic region. The level of inducing activity determines at least the time at which marginal zone cells will begin gastrulation movements. The time of its initiation of gastrulation may determine how anterior and dorsal a particular marginal zone cell can become.     

Targeting G Protein-Coupled Receptors in the Treatment of Parkinson’s Disease

Parkinson’s disease (PD) is a progressive neurodegenerative disease characterized in part by the deterioration of dopaminergic neurons which leads to motor impairment. Although there is no cure for PD, the motor symptoms can be treated using dopamine replacement therapies including the dopamine precursor L-DOPA, which has been in use since the 1960s. However, neurodegeneration in PD is not limited to dopaminergic neurons, and many patients experience non-motor symptoms including cognitive impairment or neuropsychiatric disturbances, for which there are limited treatment options. Moreover, there are currently no treatments able to alter the progression of neurodegeneration. There are many therapeutic strategies being investigated for PD, including alternatives to L-DOPA for the treatment of motor impairment, symptomatic treatments for non-motor symptoms, and neuroprotective or disease-modifying agents. G protein-coupled receptors (GPCRs), which include the dopamine receptors, are highly druggable cell surface proteins which can regulate numerous intracellular signaling pathways and thereby modulate the function of neuronal circuits affected by PD. This review will describe the treatment strategies being investigated for PD that target GPCRs and their downstream signaling mechanisms. First, we discuss new developments in dopaminergic agents for alleviating PD motor impairment, the role of dopamine receptors in L-DOPA induced dyskinesia, as well as agents targeting non-dopamine GPCRs which could augment or replace traditional dopaminergic treatments. We then discuss GPCRs as prospective treatments for neuropsychiatric and cognitive symptoms in PD. Finally, we discuss the evidence pertaining to ghrelin receptors, β-adrenergic receptors, angiotensin receptors and glucagon-like peptide 1 receptors, which have been proposed as disease modifying targets with potential neuroprotective effects in PD.     

Effect of glucose on amphetamine-induced motor behavior

The effect of 1 g/kg D-glucose on amphetamine-induced increases in activity; stereotypy; and rotation in rats with unilateral 6-OHDA lesions of nigro-striatal neurons was examined. The amphetamine dose-response curve for stereotypy was shifted to the right by glucose but retained the same slope as the control curve, suggesting that glucose competes for amphetamine or dopamine receptors that mediate this behavior. The slope of the dose-response curve for activity was significantly decreased by glucose, suggesting that glucose reduces activity levels in a manner that is non-competitive with the action of amphetamine or dopamine. Amphetamine-induced turning was unaffected by glucose at all doses tested. These data suggest that the effect of amphetamine on each of the three motor behaviors may be mediated by different dopamine-containing neurons, or by different post-synaptic dopamine receptors. The findings are also consistent with other data suggesting that glucose antagonizes dopamine function, but they suggest that this antagonism may not be uniform over all dopaminergic neurons or receptor types.     

Quantitative assessment of heterogeneous 3H-spiperone binding to rat neostriatum and frontal cortex

Anomalies of the binding of ³Hspiperone to rat cerebral membranes have been examined. By employing a very low ligand concentration ($\text{～ 25}\text{pM}{}^3\text{Hspiperone}$) we have demonstrated that even within the corpus striatum, ³Hspiperone appears to bind to multiple sites and that dopaminergic and serotonergic agents can selectively inhibit from these sites. In the corpus striatum, 75–80% of the ³Hspiperone specific binding can be inhibited with high affinity by dopaminergic drugs while some 20–30% is inhibited with high affinity by serotonergic compounds. The two ³Hspiperone sites, which we have shown to have affinities of 31 and 325 pM, may therefore represent dopaminergic and serotonergic sites. At higher concentrations of ³Hspiperone, however, the picture may be complicated by a further low affinity site. The great selectivity shown by dopaminergic agonists for the two ³Hspiperone sites explains the ‘flattened’ displacement curves reported for ³Hspiperone/agonist interactions. As dopaminergic agents show the greater affinity for the high affinity ³Hspiperone site, it is tempting to speculate that this site has the greatest association with the dopamine receptor.     

Inhibition of growth hormone excess reduces insulin resistance and ovarian dysfunction in a lean case of polycystic ovary syndrome with a growth-hormone-producing pituitary adenoma

A 23-year-old female with polycystic ovary syndrome (PCOS) and a growth-hormone (GH)-producing pituitary adenoma is described. A reduction in the elevated GH levels to normal levels following the administration of dopaminergic agents decreased plasma insulin-like growth factor (IGF)-1 and ovarian dysfunction. Menstrual cycles were therefore restored and the number of ovarian cysts reduced, suggesting that insulin and/or IGF-1, stimulators of theca cell proliferation, may be pathogenetic factors in PCOS.     

L-dopa mediated accumulation of cyclic AMP in isolated rabbit retinae in vitro. Effects of light and/or pharmacological factors

The ability of L-dopa to stimulate the formation of cyclic AMP in pieces of intact rabbit retina in vitro has been studied and compared with that of dopamine and of other dopamine-mimetic drugs. Dose-dependent effects were measured in response to 5 μM up to 100 μM L-dopa with a miximal stimulation after 20 min of incubation at 35°C. The L-dopa precursor, tyrosine, was totally ineffective. The L-dopa mediated response was detectable only in pieces of intact tissues (and not in homogenates) and is presumably due to the formation of newly formed dopamine, since it was completely inhibited by a decarboxylase inhibitor (benserazide). The biochemical response (cyclic AMP increase) was facilitated by ambient light, this effect being potentiated by 56 mM K⁺ of fully inhibited by 1 mM γ-butyrolactone (GBL). The data suggest that the measurement of cyclic AMP levels in pieces of rabbit retina may provide a useful neurochemical model for the study of physiological and/or pharmacological agents able to interact at pre- and/or post-synaptic dopaminergic sites.     

Primary Culture of Mouse Dopaminergic Neurons

Dopaminergic neurons represent less than 1% of the total number of neurons in the brain. This low amount of neurons regulates important brain functions such as motor control, motivation, and working memory. Nigrostriatal dopaminergic neurons selectively degenerate in Parkinson''s disease (PD). This progressive neuronal loss is unequivocally associated with the motors symptoms of the pathology (bradykinesia, resting tremor, and muscular rigidity). The main agent responsible of dopaminergic neuron degeneration is still unknown. However, these neurons appear to be extremely vulnerable in diverse conditions. Primary cultures constitute one of the most relevant models to investigate properties and characteristics of dopaminergic neurons. These cultures can be submitted to various stress agents that mimic PD pathology and to neuroprotective compounds in order to stop or slow down neuronal degeneration. The numerous transgenic mouse models of PD that have been generated during the last decade further increased the interest of researchers for dopaminergic neuron cultures. Here, the video protocol focuses on the delicate dissection of embryonic mouse brains. Precise excision of ventral mesencephalon is crucial to obtain neuronal cultures sufficiently rich in dopaminergic cells to allow subsequent studies. This protocol can be realized with embryonic transgenic mice and is suitable for immunofluorescence staining, quantitative PCR, second messenger quantification, or neuronal death/survival assessment.     

Evidence that serotonin neurons stimulate secretion of prolaction releasing factor.

The purpose of the present study was to determine if serotonin was stimulatory to prolactin release by inhibition of the dopaminergic system or by stimulating release of a prolactin releasing factor (PRF). We measured the amount of prolactin secreted after administration of 30 mg/kg of 5-hydroxytryptophan (5-HTP) to male rats pretreated with fluoxetine (10 mg/kg) and compared it with the amount of prolactin released in male rats treated with αmethyl-p-tyrosine methyl ester (αMT) or various dopamine receptor blocking agents. In every experiment the serotonergic stimulus provided by 5-HTP in fluoxetine-pretreated rats released considerably more prolactin than did treatment with αMT or dopaminergic blockers. We conclude that serotonin releases prolactin not by inhibiting dopaminergic neurons but rather by stimulating the release of PRF.     

Stabilized structure from motion without disparity induces disparity adaptation

3D structures can be perceived based on the patterns of 2D motion signals. With orthographic projection of a 3D stimulus onto a 2D plane, the kinetic information can give a vivid impression of depth, but the depth order is intrinsically ambiguous, resulting in bistable or even multistable interpretations. For example, an orthographic projection of dots on the surface of a rotating cylinder is perceived as a rotating cylinder with ambiguous direction of rotation. We show that the bistable rotation can be stabilized by adding information, not to the dots themselves, but to their spatial context. More interestingly, the stabilized bistable motion can generate consistent rotation aftereffects. The rotation aftereffect can only be observed when the adapting and test stimuli are presented at the same stereo depth and the same retinal location, and it is not due to attentional tracking. The observed rotation aftereffect is likely due to direction-contingent disparity adaptation, implying that stimuli with kinetic depth may have activated neurons sensitive to different disparities, even though the stimuli have zero relative disparity. Stereo depth and kinetic depth may be supported by a common neural mechanism at an early stage in the visual system.     

Effects of pimozide and domperidone administration on prolactin levels in neonatally estrogenized female rats

The effects of two dopaminergic blockers, pimozide and domperidone, on the prolactin secretion were investigated in adult female rats treated neonatally with estrogens (100 micrograms of estradiol benzoate s.c. on day 1). These rats showed hyperprolactinemia (556 micrograms/l vs 57.7 in oil-injected) and treatment with pimozide or domperidone failed to increase prolactin levels in the adult age. These results suggest that the hyperprolactinemia in neonatally estrogenized female rats is produced by loss of the dopaminergic inhibition on prolactin secretion, so that the pharmacological blockade of dopaminergic receptors is uneffective. The dopamine levels in hypothalamus were similar in control and estrogenized females suggesting that failure in dopaminergic inhibition is due to a decrease in dopamine secretion to portal vessels.     

Chronic administration of N-methyl-D-aspartate (NMDA) receptor antagonists induced in rats, a facilitation of striatal dopaminergic type D2 transmission: behavioral and biochemical study]

Compared with control rats, rats treated with ketamine (15 mg/kg/day, p.o.) or MK-801 (0.1 to 0.4 mg/kg/day, p.o.) for 6 weeks showed significantly increased: 1) behavioural responses to D2 dopaminergic agents, either agonist or antagonist, 2) striatal D2 receptor mRNA expression, 3) striatal D2 receptor density, without any presynaptic change in dopaminergic or serotoninergic neurotransmission. These results suggest that the functional expression of striatal D2 receptor is postsynaptically regulated by glutamate-triggered events through the NMDA receptor subtype.     

Evaluation of Estrogen Neuroprotective Effect on Nigrostriatal Dopaminergic Neurons Following 6-Hydroxydopamine Injection into the Substantia Nigra Pars Compacta or the Medial Forebrain Bundle

Studies involving estrogen treatment of ovariectomized rats or mice have attributed to this hormone a neuroprotective effect on the substantia nigra pars compacta (SNpc) neurons. We investigated the effect of estradiol replacement in ovariectomized rats on the survival of dopaminergic mesencephalic cell and the integrity of their projections to the striatum after microinjections of 1 microg of 6-hydroxydopamine (6-OHDA) into the right SNpc or medial forebrain bundle (MFB). Estradiol replacement did not prevent the reduction either in the striatal concentrations of DA and metabolites or in the number of nigrostriatal dopaminergic neurons following lesion with 1 microg of 6-OHDA into the SNpc. Nevertheless, estradiol treatment reduced the decrease in striatal DA following injection of 1 microg of 6-OHDA into the MFB. Results suggest therefore that estrogen protect nigrostriatal dopaminergic neurons against a 6-OHDA injury to the MFB but not the SNpc. This may be due to the distinct degree of lesions promoted in these different rat models of Parkinson's disease.