RESUMO
The D2 dopamine receptor (DRD2) is a therapeutic target for Parkinson's disease1 and antipsychotic drugs2. DRD2 is activated by the endogenous neurotransmitter dopamine and synthetic agonist drugs such as bromocriptine3, leading to stimulation of Gi and inhibition of adenylyl cyclase. Here we used cryo-electron microscopy to elucidate the structure of an agonist-bound activated DRD2-Gi complex reconstituted into a phospholipid membrane. The extracellular ligand-binding site of DRD2 is remodelled in response to agonist binding, with conformational changes in extracellular loop 2, transmembrane domain 5 (TM5), TM6 and TM7, propagating to opening of the intracellular Gi-binding site. The DRD2-Gi structure represents, to our knowledge, the first experimental model of a G-protein-coupled receptor-G-protein complex embedded in a phospholipid bilayer, which serves as a benchmark to validate the interactions seen in previous detergent-bound structures. The structure also reveals interactions that are unique to the membrane-embedded complex, including helix 8 burial in the inner leaflet, ordered lysine and arginine side chains in the membrane interfacial regions, and lipid anchoring of the G protein in the membrane. Our model of the activated DRD2 will help to inform the design of subtype-selective DRD2 ligands for multiple human central nervous system disorders.
Assuntos
Microscopia Crioeletrônica , Subunidades alfa Gi-Go de Proteínas de Ligação ao GTP/química , Subunidades alfa Gi-Go de Proteínas de Ligação ao GTP/ultraestrutura , Lipídeos de Membrana/metabolismo , Membranas Artificiais , Receptores de Dopamina D2/química , Receptores de Dopamina D2/ultraestrutura , Bromocriptina/química , Bromocriptina/metabolismo , Dopamina/química , Dopamina/metabolismo , Subunidades alfa Gi-Go de Proteínas de Ligação ao GTP/metabolismo , Humanos , Lipídeos de Membrana/química , Modelos Moleculares , Conformação Proteica , Receptores de Dopamina D2/agonistas , Receptores de Dopamina D2/metabolismo , Transdução de SinaisRESUMO
Dopaminergic medication for Parkinson's disease is associated with troubling dystonia and dyskinesia and, in rodents, dopaminergic agonists likewise induce a variety of orofacial motor responses, certain of which are mimicked by serotonin2C (5-HT2C) receptor agonists. However, the neural substrates underlying these communalities and their interrelationship remain unclear. In Sprague-Dawley rats, the dopaminergic agonist, apomorphine (0.03-0.3 mg/kg) and the preferential D2/3 receptor agonist quinpirole (0.2-0.5 mg/kg), induced purposeless oral movements (chewing, jaw tremor, tongue darting). The 5-HT2C receptor antagonist 5-methyl-1-[[2-[(2-methyl-3-pyridyl)oxyl]-5-pyridyl]carbamoyl]-6-trifluoromethylindone (SB 243213) (1 mg/kg) reduced the oral responses elicited by specific doses of both agonists (0.1 mg/kg apomorphine; 0.5 mg/kg quinpirole). After having confirmed that the oral bouts induced by quinpirole 0.5 mg/kg were blocked by another 5-HT2C antagonist (6-chloro-5-methyl-1-[6-(2-methylpiridin-3-yloxy)pyridine-3-yl carbamoyl] indoline (SB 242084), 1 mg/kg), we mapped the changes in neuronal activity in numerous sub-territories of the basal ganglia using c-Fos expression. We found a marked increase of c-Fos expression in the subthalamic nucleus (STN) in combining quinpirole (0.5 mg/kg) with either SB 243213 or SB 242084. In a parallel set of electrophysiological experiments, the same combination of SB 243213/quinpirole produced an irregular pattern of discharge and an increase in the firing rate of STN neurons. Finally, it was shown that upon the electrical stimulation of the anterior cingulate cortex, quinpirole (0.5 mg/kg) increased the response of substantia nigra pars reticulata neurons corresponding to activation of the "hyperdirect" (cortico-subthalamonigral) pathway. This effect of quinpirole was abolished by the two 5-HT2C antagonists. Collectively, these results suggest that induction of orofacial motor responses by D2/3 receptor stimulation involves 5-HT2C receptor-mediated activation of the STN by recruitment of the hyperdirect (cortico-subthalamonigral) pathway.
Assuntos
Agonistas de Dopamina/farmacologia , Receptor 5-HT2C de Serotonina/metabolismo , Serotonina/metabolismo , Núcleo Subtalâmico/efeitos dos fármacos , Aminopiridinas/farmacologia , Animais , Apomorfina/farmacologia , Gânglios da Base/efeitos dos fármacos , Gânglios da Base/metabolismo , Giro do Cíngulo/efeitos dos fármacos , Giro do Cíngulo/metabolismo , Indóis/farmacologia , Masculino , Córtex Motor/efeitos dos fármacos , Córtex Motor/metabolismo , Vias Neurais/efeitos dos fármacos , Vias Neurais/metabolismo , Proteínas Proto-Oncogênicas c-fos/metabolismo , Piridinas/farmacologia , Quimpirol/farmacologia , Ratos , Ratos Sprague-Dawley , Receptores de Dopamina D2/metabolismo , Agonistas do Receptor 5-HT2 de Serotonina/farmacologia , Antagonistas do Receptor 5-HT2 de Serotonina/farmacologia , Substância Negra/efeitos dos fármacos , Substância Negra/metabolismo , Núcleo Subtalâmico/metabolismoRESUMO
Zebrafish (Danio rerio) are ideal model organisms for investigating nervous system function, both in health and disease. Nevertheless, functional characteristics of dopamine (DA) release and uptake regulation are still not well-understood in zebrafish. In this study, we assessed D3 autoreceptor function in the telencephalon of whole zebrafish brains ex vivo by measuring the electrically stimulated DA release ([DA]max) and uptake at carbon fiber microelectrodes with fast-scan cyclic voltammetry. Treatment with pramipexole and 7-OH-DPAT, selective D3 autoreceptor agonists, sharply decreased [DA]max. Conversely, SB277011A, a selective D3 antagonist, nearly doubled [DA]max and decreased k, the first-order rate constant for the DA uptake, to about 20% of its original value. Treatment with desipramine, a selective norepinephrine transporter blocker, failed to increase current, suggesting that our electrochemical signal arises solely from the release of DA. Furthermore, blockage of DA uptake with nomifensine-reversed 7-OH-DPAT induced decreases in [DA]max. Collectively, our data show that, as in mammals, D3 autoreceptors regulate DA release, likely by inhibiting uptake. The results of this study are useful in the further development of zebrafish as a model organism for DA-related neurological disorders such as Parkinson's disease, schizophrenia, and drug addiction.
Assuntos
Autorreceptores , Peixe-Zebra , Animais , Autorreceptores/metabolismo , Encéfalo/metabolismo , Fibra de Carbono , Desipramina , Dopamina , Estimulação Elétrica , Mamíferos/metabolismo , Nomifensina , Proteínas da Membrana Plasmática de Transporte de Norepinefrina , Pramipexol , Receptores de Dopamina D2/metabolismo , Tetra-Hidronaftalenos , Peixe-Zebra/metabolismoRESUMO
Dopamine D2 receptors (D2Rs) are major targets in the treatment of psychiatric and neurodegenerative diseases. As with many other G protein-coupled receptors (GPCRs), D2Rs interact within the cellular membrane, leading to a transient receptor homo- or heterodimerization. These interactions are known to alter ligand binding, signaling, and receptor trafficking. Bivalent ligands are ideally suited to target GPCR dimers and are composed of two pharmacophores connected by a spacer element. If properly designed, bivalent ligands are able to engange the two orthosteric binding sites of a GPCR dimer simultaneously. Taking advantage of previously developed ligands for heterodimers of D2R and the neurotensin receptor 1 (NTSR1), we synthesized homobivalent ligands targeting D2R. Employing bioluminescence resonance energy transfer, we found that the bivalent ligands 3b and 4b comprising a 92-atom spacer are able to foster D2R-homodimerization while simultaneously reducing interactions of D2R with NTSR1. Both receptors are coexpressed in the central nervous system and involved in important physiological processes. The newly developed bivalent ligands are excellent tools to further understand the pharmacological consequences of D2R homo- and heterodimerization. Not limited to the dopaminergic system, modifying class A GPCRs' dynamic equilibrium between monomers, homomers, and heteromers with bivalent ligands may represent a novel pharmacological concept paving the way toward innovative drugs.
Assuntos
Agonistas de Dopamina/farmacologia , Antagonistas dos Receptores de Dopamina D2/farmacologia , Polietilenoglicóis/farmacologia , Multimerização Proteica/efeitos dos fármacos , Receptores de Dopamina D2/metabolismo , Agonistas de Dopamina/síntese química , Antagonistas dos Receptores de Dopamina D2/síntese química , Células HEK293 , Humanos , Indanos/síntese química , Indanos/farmacologia , Ligantes , Piperazinas/síntese química , Piperazinas/farmacologia , Polietilenoglicóis/síntese químicaRESUMO
Parkinson's disease is a progressive neurodegenerative disorder in which dopaminergic neurons located in the substantia nigra are gradually lost. Currently, combined treatment strategies are receiving increasing attention as potential therapeutic approaches for Parkinson's disease. This study aimed to evaluate the potential effects of exosomes released from SH-Sy5y cells and the liposomal form of L-dopa on Parkinson's rat models. Twenty-five male Wistar albino rats, in five groups, were included in this study. Parkinson's disease was induced through microinjection of 6-OHDA (2.5 mg/mL) into the right substantia nigra. The exosomes released from the SH-Sy5y cell line were isolated and administered (0.2 µg/5 µL) alone or in combination with the liposomal form of L-Dopa (80 mg/kg) to the defined model groups. Behavioral tests and molecular assays were conducted to evaluate the expression levels of tyrosine hydroxylase (TH) and dopamine receptor D2 (DRD2). The rats in the groups receiving the combined liposomal form of L-Dopa and exosome treatment and the liposomal form of L-Dopa alone showed a significant improvement in their movement ability (p < 0.05). At molecular levels, these two groups also exhibited significant increases in Th (0.005 ± 0.001) and Drd2 (0.002 ± 0.0001) expression compared to controls (p < 0.05). The observed alterations of Th and Drd2 expression were not statistically significant in exosome- and L-Dopa-treated groups. The current study shows that exosome-derived neuronal cells and liposomal form of L-Dopa can protect different cells against pathological complications such as Parkinson's disease.
Assuntos
Antiparkinsonianos/uso terapêutico , Exossomos/metabolismo , Levodopa/uso terapêutico , Doença de Parkinson/tratamento farmacológico , Receptores de Dopamina D2/metabolismo , Tirosina 3-Mono-Oxigenase/metabolismo , Animais , Antiparkinsonianos/administração & dosagem , Antiparkinsonianos/farmacologia , Linhagem Celular Tumoral , Humanos , Levodopa/administração & dosagem , Levodopa/farmacologia , Lipossomos/química , Masculino , Oxidopamina/toxicidade , Doença de Parkinson/etiologia , Doença de Parkinson/metabolismo , Ratos , Ratos Wistar , Transdução de Sinais , Substância Negra/efeitos dos fármacos , Substância Negra/metabolismoRESUMO
Converging evidence suggests that oxytocin (OT) is associated with creative thinking (CT) and that release of OT depends on ADP ribosyl-cyclases (CD38 and CD157). Neural mechanisms of CT and OT show a strong association with dopaminergic (DA) pathways, yet the link between CT and CD38, CD157, dopamine receptor D2 (DRD2) and catechol-O-methyltransferase (COMT) peripheral gene expression remain inconclusive, thus limiting our understanding of the neurobiology of CT. To address this issue, two principal domains of CT, divergent thinking (AUT), were assessed. In men, both AUT is associated with gene expression of CD38, CD157, and their interaction CD38 × CD157. There were no significant associations for DA expression (DRD2, COMT, DRD2 × COMT) on both CT measures. However, analysis of the interactions of OT and DA systems reveal significant interactions for AUT in men. The full model explained a sizable 39% of the variance in females for the total CT score. The current findings suggest that OT and DA gene expression contributed significantly to cognition and CT phenotype. This provides the first empirical foundation of a more refined understanding of the molecular landscape of CT.
Assuntos
Cognição/efeitos dos fármacos , Criatividade , Dopamina/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Ocitocina/farmacologia , Saliva/metabolismo , ADP-Ribosil Ciclase/genética , ADP-Ribosil Ciclase/metabolismo , ADP-Ribosil Ciclase 1/genética , ADP-Ribosil Ciclase 1/metabolismo , Adulto , Antígenos CD/genética , Antígenos CD/metabolismo , Catecol O-Metiltransferase/genética , Catecol O-Metiltransferase/metabolismo , Dopaminérgicos/farmacologia , Feminino , Proteínas Ligadas por GPI/genética , Proteínas Ligadas por GPI/metabolismo , Interação Gene-Ambiente , Humanos , Masculino , Glicoproteínas de Membrana/genética , Glicoproteínas de Membrana/metabolismo , Ocitócicos/farmacologia , Polimorfismo de Nucleotídeo Único , Receptores de Dopamina D2/genética , Receptores de Dopamina D2/metabolismo , Saliva/efeitos dos fármacos , Fatores Sexuais , Adulto JovemRESUMO
Currently, there is mounting evidence that intermolecular receptor-receptor interactions may result in altered receptor recognition, pharmacology and signaling. Heterobivalent ligands have been proven useful as molecular probes for confirming and targeting heteromeric receptors. This report describes the design and synthesis of novel heterobivalent ligands for dopamine D2 -like receptors (D2 -likeR) and the µ-opioid receptor (µOR) and their evaluation using ligand binding and functional assays. Interestingly, we identified a potent bivalent ligand that contains a short 18-atom linker and combines good potency with high efficacy both in ß-arrestinâ 2 recruitment for µOR and MAPK-P for D4 R. Furthermore, this compound was characterized by a biphasic competition binding curve for the D4 R-µOR heterodimer, indicative of a bivalent binding mode. As this compound possibly bridges the D4 R-µOR heterodimer, it could be used as a pharmacological tool to further investigate the interactions of D4 R and µOR.
Assuntos
Desenho de Fármacos , Sondas Moleculares/farmacologia , Polietilenoglicóis/farmacologia , Receptores de Dopamina D2/agonistas , Receptores Opioides mu/agonistas , Receptores Opioides mu/antagonistas & inibidores , Células Cultivadas , Relação Dose-Resposta a Droga , Células HEK293 , Humanos , Ligantes , Sondas Moleculares/síntese química , Sondas Moleculares/química , Estrutura Molecular , Polietilenoglicóis/síntese química , Polietilenoglicóis/química , Receptores de Dopamina D2/metabolismo , Receptores Opioides mu/metabolismo , Relação Estrutura-AtividadeRESUMO
Though orofacial movements are fundamental motor patterns that are known to be regulated critically by D1-like dopamine receptors, these processes remain poorly understood. This uncertainty is heightened by evidence for putative D1-like receptors that are linked not only to adenylyl cyclase (AC) but also to phospholipase C (PLC). Using a new method, we have characterised four topographies of orofacial movement in the mouse using the novel D1-like agonist SKF 83822, which stimulates AC but not PLC. These were compared with responses to SKF 83959, which stimulates PLC but not AC. Also, effects were characterised using the D1-like antagonist SCH 23390 and the D2-like antagonist YM 09151-2. SKF 83822 induced vertical jaw movements with incisor chattering but inhibited horizontal jaw movements; there was little effect on tongue protrusions. Vertical jaw movements induced by SKF 83822 were inhibited by SCH 23390 but uninfluenced by YM 09151-2, while YM 09151-2 released horizontal jaw movements; thus, D1-like agonist-induced, AC-mediated vertical jaw movements constitute a 'pure' D1-like-dependent process that does not involve D1-like:D2-like interactions, while horizontal jaw movements involve oppositional interactions. Orofacial movements in mice appear to consist of at least four phenomenologically dissociable topographies that are mechanistically distinct. They are regulated differentially by AC- and/or PLC-dependent processes and these processes involve distinct D1-like:D2-like interactions.
Assuntos
Adenilil Ciclases/metabolismo , Músculos Faciais/fisiologia , Músculos da Mastigação/fisiologia , Movimento/fisiologia , Receptores de Dopamina D1/metabolismo , Fosfolipases Tipo C/metabolismo , Animais , Benzazepinas/farmacologia , Mapeamento Encefálico , Dopamina/metabolismo , Agonistas de Dopamina/farmacologia , Antagonistas de Dopamina/farmacologia , Antagonistas dos Receptores de Dopamina D2 , Relação Dose-Resposta a Droga , Vias Eferentes/efeitos dos fármacos , Vias Eferentes/fisiologia , Músculos Faciais/inervação , Masculino , Músculos da Mastigação/inervação , Camundongos , Camundongos Endogâmicos C57BL , Córtex Motor/efeitos dos fármacos , Córtex Motor/fisiologia , Movimento/efeitos dos fármacos , Rede Nervosa/efeitos dos fármacos , Rede Nervosa/fisiologia , Receptores de Dopamina D1/agonistas , Receptores de Dopamina D1/antagonistas & inibidores , Receptores de Dopamina D2/agonistas , Receptores de Dopamina D2/metabolismo , Transmissão Sináptica/efeitos dos fármacos , Transmissão Sináptica/fisiologiaRESUMO
In order to better define the role of 5-HT(1A) receptors in the modulation of extrapyramidal motor functions, we investigated the effect of 5-HT(1A) agonists on tacrine-induced tremulous jaw movements (TJM) in rats, a putative model of parkinsonian tremor. Acute injection of 5-HT(1A) agonists 8-OH-DPAT and buspirone dose-dependently counteracted the tacrine-induced oral movements (ED(50)=0.04 and 1.0mg/kg, respectively), an effect reversed by the selective 5-HT(1A) antagonist WAY 100,635. In contrast to classical antipsychotics, the atypical antipsychotics risperidone (ED(50)=0.3mg/kg) and clozapine (ED(50)=1.5mg/kg) blocked the oral movements induced by the cholinomimetic agent at or below the doses required for suppression of conditioned avoidance response. The compound F-97013-GD (6-methyl-2-[4-(naphtylpiperazin-1-yl)butyl]-3-(2H)-pyridazinone), a putative antipsychotic drug that in functional in vitro and in vivo assays behaved as a mixed dopamine D(2)-antagonist and 5-HT(1A)-partial agonist, also displayed a potent antitremorgenic effect in this paradigm (ED(50)=0.5mg/kg). Interestingly, pretreatment with WAY 100,635 blocked the inhibitory effect of F-97013-GD but not that of clozapine. The 5-HT depleting agent para-chlorophenylalanine (PCPA) partially attenuated tacrine-induced TJM but did not block the suppressive effect of 5-HT(1A) agonists. In addition, only high doses of F-97013-GD induced catalepsy in rodents and, like 8-OH-DPAT and clozapine, the compound reversed the haloperidol-induced catalepsy in rats. These results show that 5-HT(1A) receptors play a role in the regulation of tacrine-induced TJM and suggest that their activation by novel antipsychotics may not only reduce the extrapyramidal side effects EPS liability, but also be effective in the treatment of parkinsonian tremor.
Assuntos
Antiparkinsonianos/farmacologia , Antipsicóticos/farmacologia , Piperazinas/farmacologia , Piridazinas/farmacologia , Receptor 5-HT1A de Serotonina/efeitos dos fármacos , Agonistas do Receptor de Serotonina/farmacologia , 5-Hidroxitriptofano/farmacologia , Animais , Apomorfina/farmacologia , Aprendizagem da Esquiva/efeitos dos fármacos , Ligação Competitiva/efeitos dos fármacos , Catalepsia/induzido quimicamente , Agonistas de Dopamina/farmacologia , Antagonistas dos Receptores de Dopamina D2 , Proteínas da Membrana Plasmática de Transporte de Dopamina/metabolismo , Interações Medicamentosas , Guanosina 5'-O-(3-Tiotrifosfato)/metabolismo , Haloperidol/antagonistas & inibidores , Movimentos da Cabeça/efeitos dos fármacos , Hipotermia/induzido quimicamente , Hipotermia/fisiopatologia , Arcada Osseodentária/fisiologia , Masculino , Camundongos , Atividade Motora/efeitos dos fármacos , Piperazinas/metabolismo , Piridinas/metabolismo , Ratos , Ratos Wistar , Receptores de Dopamina D2/metabolismo , Antagonistas da Serotonina/metabolismo , Tremor/fisiopatologiaRESUMO
Mesolimbic dopamine is thought to play a role in the processing of rewards. However, animal studies also demonstrate dopamine release in response to aversive stressful stimuli. Also, in animal studies, disruptions of the mother-infant relationship have been shown to have long-lasting effects on the mesolimbic dopamine system and the hypothalamic-pituitary adrenal axis. We therefore investigated dopamine release in response to stress in human subjects, considering the relationship to early life parental care. We screened 120 healthy young college students for parental care in early life using a combination of telephone interviews and questionnaires. Five students from the top end and five students from the bottom end of the parental care distribution were then invited for a positron emission tomography study using [11C]raclopride and a psychosocial stress task. The psychosocial stressor caused a significant release of dopamine in the ventral striatum as indicated by a reduction in [11C]raclopride binding potential in the stress versus resting condition in subjects reporting low parental care. Moreover, the magnitude of the salivary cortisol response to stress was significantly correlated with the reduction in [11C]raclopride binding in the ventral striatum (r = 0.78), consistent with a facilitating effect of cortisol on dopamine neuron firing. These data suggest that aversive stressful events can be associated with mesolimbic dopamine release in humans, and that the method presented here may be useful to study the effects of early life events on neurobiological stress systems.
Assuntos
Encéfalo/metabolismo , Dopamina/metabolismo , Comportamento Materno/psicologia , Racloprida/metabolismo , Estresse Psicológico/metabolismo , Estresse Psicológico/psicologia , Adulto , Gânglios da Base/diagnóstico por imagem , Gânglios da Base/metabolismo , Ligação Competitiva/fisiologia , Encéfalo/diagnóstico por imagem , Antagonistas de Dopamina/metabolismo , Antagonistas de Dopamina/farmacocinética , Feminino , Humanos , Hidrocortisona/análise , Hidrocortisona/metabolismo , Masculino , Apego ao Objeto , Resolução de Problemas , Testes Psicológicos , Racloprida/farmacocinética , Receptores de Dopamina D2/metabolismo , Valores de Referência , Saliva/química , Tomografia Computadorizada de EmissãoRESUMO
Animal studies suggest that the dopaminergic system plays a role in central pain modulation. We have previously demonstrated with positron emission tomography (PET) that striatal dopaminergic hypofunction may be involved in the burning mouth syndrome. The aim of the present study was to evaluate the nigrostriatal dopaminergic system in patients with atypical facial pain using PET. In seven patients with atypical facial pain, striatal presynaptic dopaminergic function was assessed with [18F]FDOPA and dopamine D1 and D2 receptor availabilities with [11C]NNC 756 and [11C]raclopride, respectively. The results were compared with those of healthy controls. A quantitative region-of-interest analysis showed that the uptakes of [18F]FDOPA and [11C]NNC 756 did not differ between patients and controls. There was a tendency of increased D2 receptor availability in the left putamen (P=0.056), and the D1/D2 ratio in the putamen was decreased bilaterally by 7.7% (P=0.002) in patients when compared to controls. In a voxel-based analysis, the uptake of [11C]raclopride was increased in the left putamen (P=0.025). In conclusion, the increase in D2 receptor availability in the left putamen and the decrease in D1/D2 ratio imply that alterations in the striatal dopaminergic system as evaluated by PET may be involved in chronic orofacial pain conditions.
Assuntos
Di-Hidroxifenilalanina/análogos & derivados , Dor Facial/diagnóstico por imagem , Dor Facial/metabolismo , Receptores de Dopamina D2/metabolismo , Idoso , Benzazepinas , Benzofuranos , Radioisótopos de Carbono , Antagonistas de Dopamina , Humanos , Pessoa de Meia-Idade , Putamen/metabolismo , Racloprida , Receptores de Dopamina D1/metabolismo , Tomografia Computadorizada de EmissãoRESUMO
Tardive dyskinesia develops as a common complication of long-term neuroleptic use. The emergence of such dyskinesias may reflect a shift in the balance of dopamine D(1) and D(2) receptor-mediated activity, with a relative increase in activity in the D(1) receptor-regulated direct striatonigral pathway. In rats, chronic treatment with the antipsychotic fluphenazine triggers a syndrome of vacuous chewing movements, which are attenuated by dopamine D(1) receptor antagonists. A similar syndrome can be seen in drug-naive animals following acute administration of selective dopamine D(1) receptor agonists. However, not all dopamine D(1) receptor agonists elicit these mouth movements. Thus, some investigators have suggested the existence of novel subtypes of the dopamine D(1) receptor. In these studies, we sought to clarify the role of the dopamine D(1A) receptor in vacuous chewing movements induced both by the selective dopamine D(1) receptor agonist SKF 38393, as well as by chronic neuroleptic administration, using in vivo oligonucleotide antisense to dopamine D(1A) receptor messenger RNA. Intrastriatal antisense treatment significantly and selectively attenuated striatal dopamine D(1) receptor binding, accompanied by reductions in SKF 38393- and chronic fluphenazine-induced vacuous chewing movements. These findings suggest that the dopamine D(1A) receptor plays an important role in the expression of vacuous chewing movements in a rodent model of tardive dyskinesia and may contribute to the pathogenesis of the human disorder. This may have important implications for the treatment of tardive dyskinesia in humans.
Assuntos
Discinesia Induzida por Medicamentos/metabolismo , Neostriado/efeitos dos fármacos , Neurônios/efeitos dos fármacos , Receptores de Dopamina D1/efeitos dos fármacos , Receptores de Dopamina D1/metabolismo , 2,3,4,5-Tetra-Hidro-7,8-Di-Hidroxi-1-Fenil-1H-3-Benzazepina/farmacologia , Animais , Antipsicóticos/efeitos adversos , Comportamento Animal/efeitos dos fármacos , Comportamento Animal/fisiologia , Sítios de Ligação/efeitos dos fármacos , Sítios de Ligação/fisiologia , Modelos Animais de Doenças , Agonistas de Dopamina/farmacologia , Discinesia Induzida por Medicamentos/patologia , Discinesia Induzida por Medicamentos/fisiopatologia , Flufenazina/farmacologia , Masculino , Mastigação/efeitos dos fármacos , Mastigação/fisiologia , Neostriado/patologia , Neostriado/fisiopatologia , Neurônios/metabolismo , Neurônios/patologia , RNA Mensageiro/efeitos dos fármacos , RNA Mensageiro/metabolismo , Racloprida/farmacologia , Ratos , Ratos Sprague-Dawley , Receptores de Dopamina D1/genética , Receptores de Dopamina D2/efeitos dos fármacos , Receptores de Dopamina D2/metabolismo , TrítioRESUMO
The D3 dopamine receptor belongs to the D2-like family of dopamine receptors. As with other members of this group, the D3 dopamine receptor gene contains introns which allow for alternative splicing of gene products. The best characterized of the human D3 dopamine receptor mRNA splice variants encodes a truncated protein called D3nf. The D3 dopamine receptor and D3nf were epitope-tagged and expressed in Sf9 insect cells by recombinant baculovirus infection. The D3 dopamine receptor showed saturable, high affinity binding of agonists and antagonists, consistent with reported D3 dopamine receptor pharmacology. When the D3 dopamine receptor and D3nf were co-expressed, the apparent density of D3 dopamine receptor expression, as determined by radioligand binding, was significantly lowered compared to D3 dopamine receptor expressed alone. This effect of D3nf was specific for the D3 dopamine receptor, since co-expression with the D2 dopamine receptor or beta2-adrenoceptor had no effect on binding. Confocal immunofluorescence studies were used to confirm that both D3 dopamine receptor and D3nf were well expressed on the cell surface and densitometric analysis of cell surface membrane protein confirmed that D3nf did not significantly alter the amount of D3 dopamine receptor expressed. Photoaffinity labelling with [125I]azidonemonapride showed that the amount of ligand bound by membranes co-expressing D3 dopamine receptor and D3nf was significantly less than that bound by membranes expressing D3 dopamine receptor alone. The greatest decrease in binding was observed in the D3 dopamine receptor oligomeric forms. Ligand binding to dimers and tetramers was reduced by 69 and 46%, respectively, indicating effects of a protein-protein interaction. Co-immunoprecipitation confirmed that the D3DR and D3nf interact with each other. These data indicate that D3nf heterodimerizes with the D3 dopamine receptor and decreases the capacity of D3 dopamine receptor to bind ligand.
Assuntos
Processamento Alternativo , Receptores de Dopamina D2/genética , Receptores de Dopamina D2/metabolismo , Animais , Baculoviridae , Dimerização , Eletroforese em Gel de Poliacrilamida , Vetores Genéticos , Humanos , Ligantes , Microscopia Confocal , Polímeros/metabolismo , Ligação Proteica , Conformação Proteica , Receptores de Dopamina D3 , Spodoptera , Relação Estrutura-AtividadeRESUMO
The effects of repeated administration of ceruletide (100 micrograms/kg/perday, i.p. for 3 days) on perioral movements and the striatal dopamine receptor adenylate cyclase system were examined in rats chronically treated with fluphenazine enanthate (FPZ) (25 mg/kg i.m. every 3 weeks for 30 weeks) and sesame oil-treated (control) rats. After the tenth injection of fluphenazine, the rats started to display five types of perioral movements (teeth chattering, chewing, tongue protrusion, mouth opening and perioral tremors). Moreover, increases in SCH23390 binding and spiperone binding to striatal membranes, were found in the FPZ-treated rats. Furthermore, dopamine receptor-coupled adenylate cyclase activity was potentiated in striatal membranes. High amplitude EMG discharges (8-10 Hz), recorded from the masseter in the FPZ-treated rats occurred concurrently with perioral tremors. Repeated ceruletide (CLT) injections abolished perioral movements, and reversed both the elevated SCH23390 binding and the dopamine stimulated adenylate cyclase (AC) activity to the control level. The effect of CLT on perioral movements, D1 receptors and dopamine-stimulated AC activity continued for 6 days after the final CLT injection. These findings suggest that systemically administered CLT affects the D1 receptor adenylate cyclase system and that an increase of the D1 receptor mechanism may play an important role in the pathogenesis of tardive dyskinesia.
Assuntos
Adenilil Ciclases/efeitos dos fármacos , Comportamento Animal/efeitos dos fármacos , Ceruletídeo/farmacologia , Corpo Estriado/efeitos dos fármacos , Antagonistas de Dopamina/farmacologia , Flufenazina/administração & dosagem , Adenilil Ciclases/metabolismo , Animais , Benzazepinas/metabolismo , Corpo Estriado/metabolismo , Dopamina/farmacologia , Interações Medicamentosas , Eletromiografia , Masculino , Ratos , Ratos Wistar , Receptores Dopaminérgicos/efeitos dos fármacos , Receptores Dopaminérgicos/metabolismo , Receptores de Dopamina D1/efeitos dos fármacos , Receptores de Dopamina D1/metabolismo , Receptores de Dopamina D2/efeitos dos fármacos , Receptores de Dopamina D2/metabolismo , Espiperona/metabolismoRESUMO
A novel procedure for the assessment of orofacial movement topographies in mice was used to study, for the first time, the individual and interactive involvement of dopamine D(1)-like vs. D(2)-like receptors in their regulation. The dopamine D(1)-like receptor agonists A 68930 ([1R,3S]-1-aminomethyl-5,6-dihydroxy-3-phenyl-isochroman) and SK&F 83959 (3-methyl-6-chloro-7,8-dihydroxy-1-[3-methyl-phenyl]-2,3,4,5-tetrahydro-1H-3-benzazepine) each induced vertical jaw movements with tongue protrusions and incisor chattering. The dopamine D(1)-like receptor antagonists SCH 23390 ([R]-3-methyl-7-chloro-8-hydroxy-1-phenyl-2,3,4,5-tetrahydro-1H-3-benzazepine) and BW 737C ([S]-6-chloro-1-[2,5-dimethoxy-4-propylbenzyl]-7-hydroxy-2-methyl-1,2,3,4-tetrahydroisoquinoline) antagonised these responses, while the dopamine D(2)-like receptor antagonist YM 09151-2 (cis-N-[1-benzyl-2-methyl-pyrrolidin-3-yl]-5-chloro-2-methoxy-4-methylaminobenzamide) attenuated those to SK&F 83959 and released horizontal jaw movements. These findings suggest some role for a dopamine D(1)-like receptor that is coupled to a transduction system other than/additional to adenylyl cyclase, and for dopamine D(1)-like:D(2)-like receptor interactions, in the regulation of individual orofacial movement topographies in the mouse. This methodology will allow the use of knockout mice to clarify the roles of individual dopamine receptor subtypes in their regulation.
Assuntos
Face/fisiologia , Cabeça/fisiologia , Movimento , Receptores de Dopamina D1/metabolismo , Receptores de Dopamina D2/metabolismo , Animais , Comportamento Animal/efeitos dos fármacos , Agonistas de Dopamina/farmacologia , Antagonistas de Dopamina/farmacologia , Antagonistas dos Receptores de Dopamina D2 , Feminino , Arcada Osseodentária/efeitos dos fármacos , Arcada Osseodentária/fisiologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Movimento/efeitos dos fármacos , Receptores de Dopamina D1/agonistas , Receptores de Dopamina D1/antagonistas & inibidores , Receptores de Dopamina D2/agonistas , Língua/efeitos dos fármacos , Língua/fisiologia , Vibrissas/efeitos dos fármacos , Vibrissas/fisiologiaRESUMO
Bilateral injections of the thyrotropin-releasing hormone (TRH) analogue, N alpha-[((S)-4-oxo-2-azetidinyl)-carbonyl]-L-histidyl-L-prolinamide dihydrate (YM-14673, 0.1 microgram and 1 microgram/0.2 microliters), into the nucleus accumbens, the dorsal and ventrolateral striatum produced repetitive jaw movements in a dose-dependent manner. The effects were greatest in the nucleus accumbens and smallest in the ventrolateral striatum. Pattern of the movements differed from that produced by injections of a mixture of SKF 38393 (5 micrograms) and quinpirole (10 micrograms); frequent tongue protrusions were evident in rats treated with the mixture but those were not seen in YM-14673-treated rats. TRH (1 microgram, 10 micrograms and 30 micrograms/0.2 microliters) did not evoke jaw movements from any of the sites. The non-selective dopamine receptor antagonist, cis-(Z)-flupentixol (10 micrograms), significantly reduced the response to administration of YM-14673 (1 microgram) into the nucleus accumbens or dorsal striatum, while the 5-hydroxytryptamine (5-HT)2A receptor antagonist, 2-(2-dimethylaminoethylthio)-3-phenylquinoline hydrochloride (ICI 169,369, 0.2 micrograms), did not affect the response to YM-14673 (1 microgram). Given intrathecally (0.5 microgram/5 microliters), both YM-14673 and TRH produced wet-dog shakes. Although the mechanisms giving rise to the display of jaw movements after intrastriatal injections of YM-14673 remain unknown, stimulation of the dopamine D1/D2 receptors may at least partly contribute to these effects. Anyhow, these mechanisms differ from that underlying the ability of YM-14673 and TRH to elicit wet-dog shakes, a mechanism that is known to involve serotonergic processes.
Assuntos
Azetidinas/farmacologia , Corpo Estriado/efeitos dos fármacos , Dipeptídeos/farmacologia , Arcada Osseodentária/efeitos dos fármacos , Núcleo Accumbens/efeitos dos fármacos , Animais , Azetidinas/administração & dosagem , Comportamento Animal/efeitos dos fármacos , Corpo Estriado/metabolismo , Dipeptídeos/administração & dosagem , Relação Dose-Resposta a Droga , Eletromiografia , Flupentixol/administração & dosagem , Flupentixol/farmacologia , Injeções Espinhais , Arcada Osseodentária/fisiologia , Masculino , Núcleo Accumbens/metabolismo , Quinolinas/administração & dosagem , Quinolinas/farmacologia , Ratos , Ratos Sprague-Dawley , Receptores de Dopamina D1/efeitos dos fármacos , Receptores de Dopamina D1/metabolismo , Receptores de Dopamina D2/efeitos dos fármacos , Receptores de Dopamina D2/metabolismo , Antagonistas da Serotonina/administração & dosagem , Antagonistas da Serotonina/farmacologia , Hormônio Liberador de Tireotropina/análogos & derivados , Hormônio Liberador de Tireotropina/farmacologiaRESUMO
Huntington's disease (HD) results from the degeneration of striatal neurones, mainly gamma-aminobutyric acid (GABA)ergic projection neurones and lately cholinergic interneurones. The use of trophic factors as agents able to prevent such neural degeneration is a promising strategy. The aim of this study was to validate nerve growth factor-loaded (NGF-loaded) poly-D,L-lactide-co-glycolide (PLGA) microspheres for treatment of HD in a rat model with quinolinic acid lesion using autoradiographic study of D2 dopaminergic receptors (D2R). This target is expressed by about half of striatal neurones and its scintigraphic exploration has already been performed for the follow-up of this degenerative process. Ex vivo autoradiography of D2R performed with iodobenzamide, the widely used ligand for single photo emission computerized tomography, revealed slight neuroprotection. Moreover, tolerance of microspheres was demonstrated by in vitro autoradiography with the marker of gliosis, [(3)H]-PK 11195.
Assuntos
Interneurônios/efeitos dos fármacos , Fator de Crescimento Neural/farmacocinética , Fármacos Neuroprotetores/farmacocinética , Receptores de Dopamina D2/análise , Animais , Antineoplásicos/metabolismo , Antineoplásicos/farmacologia , Autorradiografia , Benzamidas/metabolismo , Benzamidas/farmacologia , Materiais Biocompatíveis , Corpo Estriado/citologia , Denervação , Antagonistas de Dopamina/metabolismo , Antagonistas de Dopamina/farmacologia , Portadores de Fármacos , Doença de Huntington/tratamento farmacológico , Interneurônios/química , Radioisótopos do Iodo , Isoquinolinas/metabolismo , Isoquinolinas/farmacologia , Ácido Láctico , Masculino , Microesferas , Ácido Poliglicólico , Copolímero de Ácido Poliláctico e Ácido Poliglicólico , Polímeros , Pirrolidinas/metabolismo , Pirrolidinas/farmacologia , Ácido Quinolínico , Ratos , Ratos Wistar , Receptores de Dopamina D2/metabolismo , TrítioRESUMO
The neurochemical mechanisms underlying sleep bruxism are little understood at present. However, recent pharmacologic evidence suggests that the central dopaminergic system may be involved in the pathophysiology of sleep bruxism. This possibility was further assessed by means of functional neuroimaging of dopamine D2 receptors with single-photon-emission computed tomography (SPECT). Ten controls and ten patients with polysomnographically confirmed sleep bruxism were injected intravenously with 185 MBq (5 mCi) iodine-123-iodobenzamide, a specific D2 receptor antagonist radioligand, and data acquisition was performed 90 min post-injection. Following image reconstruction, it was found that striatal D2 receptor binding potential (basal ganglia/background ratio) did not differ significantly between bruxism patients and controls. However, side-to-side differences between unilateral values of the striatal D2 binding potential ("highest side" values minus "lowest side" values) were significantly larger for the bruxism patients (p < 0.001, by two-independent-samples t test with pooled variances). It was concluded that an abnormal side imbalance in striatal D2 receptor expression can be associated with sleep bruxism. This reinforces the possibility that the central dopaminergic system plays a role in the pathophysiology of this disorder.
Assuntos
Benzamidas , Bruxismo/metabolismo , Corpo Estriado/metabolismo , Pirrolidinas , Receptores de Dopamina D2/metabolismo , Tomografia Computadorizada de Emissão de Fóton Único , Adulto , Bruxismo/diagnóstico por imagem , Corpo Estriado/diagnóstico por imagem , Antagonistas dos Receptores de Dopamina D2 , Feminino , Humanos , Radioisótopos do Iodo , Ligantes , Masculino , PolissonografiaRESUMO
This study was designed to assess the effects of bromocriptine, a dopamine D2 receptor agonist, on sleep bruxism. Seven otherwise healthy patients with severe and frequent sleep bruxism participated in this randomized, double-blind, placebo-controlled study. The study used a crossover design that included 2 weeks of active treatment or placebo with a washout period of 1 week. To further evaluate whether bromocriptine influences striatal D2 receptor binding, we used iodine-123-iodobenzamide single photon emission computed tomography (SPECT) under both placebo and bromocriptine regimens. Bromocriptine did not reduce the frequency of episodes of bruxism during sleep (mean +/- SEM, 9.0 +/- 1.0 and 9.6 +/- 1.5 bruxism episodes per hour for placebo and bromocriptine, respectively) or the amplitude of masseter muscle contractions (root mean square values, 48.2 +/- 15.5 microV and 46.9 +/- 12.7 microV for placebo and bromocriptine, respectively). SPECT also failed to reveal that either treatment had any influence on striatal D2 binding (values for total binding in counts/pixel, 1.80 [1.72-1.93] and 1.79 [1.56-1.87] for placebo and bromocriptine, respectively). This study shows that a nightly dose of bromocriptine does not exacerbate or reduce sleep bruxism motor activity.
Assuntos
Bromocriptina/uso terapêutico , Bruxismo/tratamento farmacológico , Agonistas de Dopamina/uso terapêutico , Adulto , Bromocriptina/efeitos adversos , Estudos Cross-Over , Agonistas de Dopamina/efeitos adversos , Método Duplo-Cego , Feminino , Humanos , Masculino , Receptores de Dopamina D2/efeitos dos fármacos , Receptores de Dopamina D2/metabolismo , Sono/efeitos dos fármacos , Fases do Sono/efeitos dos fármacosRESUMO
AIMS: First, to evaluate possible orofacial morphologic differences between sleep bruxers and non-bruxers, and second, to determine possible correlations between morphologic factors and striatal D2 receptor expression in persons with sleep-related oromotor activities. METHODS: Twenty subjects were included in this study; half of them had polysomnographically confirmed oromotor values above the cutoff points for sleep bruxism. For all participants, 26 standard occlusal measures were recorded clinically and from dental study casts. In addition, 25 standard angular and linear measures were taken from standardized cephalometric films, and variables were derived to evaluate dental and skeletal relationships. Fourteen of the 20 participants had also participated in a previous study that included iodine-123-iodobenzamide (I-123-IBZM) and single-photon emission-computed tomography (SPECT). For them, the side-to-side difference in striatal D2 receptor binding was determined as the neurochemical outcome measure. RESULTS: Following the classical Bonferroni adjustment for multiple testing, no morphologic differences were found between the sleep bruxers and the non-bruxers. In addition, none of the morphologic variables were significantly associated with the neuroimaging data. CONCLUSION: Taking into account the low power of this retrospective, exploratory study, the results suggest that the orofacial morphology of sleep bruxers does not differ from that of non-bruxers. In addition, morphologic factors are probably not involved in the asymmetry in striatal D2 receptor distribution that was previously observed in association with sleep bruxism.