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1.
Psychiatr Danub ; 31(2): 235-240, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31291231

RESUMO

BACKGROUND: Previous research showed inconsistent results concerning a possible association between solute carrier family 6 member 3 (SLC6A3) gene polymorphisms and dopamine symptoms of posttraumatic stress disorder (PTSD). Several studies also indicate that the myelin basic protein (MBP) gene is of importance in the etiology of several psychiatric disorders. The aim of this study was to investigate the relation of distinct SLC6A3 and MBP gene polymorphisms with PTSD and whether SLC6A3 and MBP genotypes contribute to PTSD symptom severity. SUBJECTS AND METHODS: The study included 719 individuals who had experienced war trauma in the South Eastern Europe (SEE). Genotypes of variable number tandem repeat (VNTR) polymorphism within the SLC6A3 gene were assessed in 696 participants, and the single nucleotide polymorphism (SNP) rs12458282 located within the MBP gene region was genotyped in a total of 703 subjects. The Mini International Neuropsychiatric Interview, the Clinical Administrated PTSD Scale (CAPS) and Brief Symptom Inventory (BSI), were used for data collection. RESULTS: No significant differences concerning the investigated SLC6A3 and MBP polymorphisms was identifiable between PTSD and non PTSD participants. Also we could not detect significant influence of these distinct SLC6A3 and MBP alleles on the severity of PTSD symptoms (CAPS) or BSI scores. However, the results of MBP rs12458282 within the patients with lifetime PTSD may point to a possible correlation of the major allele (T) with elevated CAPS scores. CONCLUSIONS: Our results do not support an association of the analysed SLC6A3 and MBP gene polymorphisms with PTSD in war traumatized individuals. We found that there is a possibility for a correlation of the T allele rs12458282 within the MBP gene with higher CAPS scores in lifetime PTSD patients which would need to be tested in a sample providing more statistical power.


Assuntos
Proteínas da Membrana Plasmática de Transporte de Dopamina/genética , Proteína Básica da Mielina/genética , Polimorfismo de Nucleotídeo Único , Transtornos de Estresse Pós-Traumáticos/genética , Alelos , Feminino , Genótipo , Humanos , Masculino , Pessoa de Meia-Idade
2.
Nat Commun ; 10(1): 2508, 2019 06 07.
Artigo em Inglês | MEDLINE | ID: mdl-31175277

RESUMO

Midbrain dopamine (mDA) neurons play a central role in reward signaling and are widely implicated in psychiatric and neurodegenerative disorders. To understand how mDA neurons perform these functions, it is important to understand how mDA-specific genes are regulated. However, cellular heterogeneity in the mammalian brain presents a major challenge to obtaining this understanding. To this end, we developed a virus-based approach to label and capture mDA nuclei for transcriptome (RNA-Seq), and low-input chromatin accessibility (liDNase-Seq) profiling, followed by predictive modeling to identify putative transcriptional regulators of mDA neurons. Using this method, we identified Gmeb1, a transcription factor predicted to regulate expression of Th and Dat, genes critical for dopamine synthesis and reuptake, respectively. Gmeb1 knockdown in mDA neurons resulted in downregulation of Th and Dat, as well as in severe motor deficits. This study thus identifies Gmeb1 as a master regulator of mDA gene expression and function, and provides a general method for identifying cell type-specific transcriptional regulators.


Assuntos
Proteínas da Membrana Plasmática de Transporte de Dopamina/genética , Neurônios Dopaminérgicos/metabolismo , Parte Compacta da Substância Negra/metabolismo , Fatores de Transcrição/genética , Tirosina 3-Mono-Oxigenase/genética , Animais , Perfilação da Expressão Gênica , Regulação da Expressão Gênica , Técnicas de Silenciamento de Genes , Mesencéfalo/citologia , Camundongos
3.
Int J Mol Sci ; 20(8)2019 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-30991630

RESUMO

Studies indicate the heritable nature of affective temperament, which shows personality traits predisposing to the development of mental disorders. Dopaminergic gene polymorphisms such as DRD4, COMTVal158Met, and DAT1 have been linked to affective disorders in obesity. Due to possible correlation between the aforementioned polymorphisms and the affective temperament, the aim of our research was to investigate this connection in an obese population. The study enrolled 245 obese patients (178 females; 67 males). The affective temperament was assessed using the Temperament Evaluation of Memphis, Pisa, Paris, and San Diego autoquestionnaire (TEMPS-A). Genetic polymorphisms of DAT1, COMTVal158Met and DRD4 were collected from peripheral blood sample and determined using a polymerase chain reaction (PCR). Only in COMT polymorphisms, the cyclothymic and irritable dimensions were significantly associated with Met/Val carriers (p = 0.04; p = 0.01). Another interesting finding was the correlation between the affective temperament and age in men and women. We assume that dopamine transmission in heterozygotes of COMT may determine the role of the affective temperament in obese persons. Dopaminergic transmission modulated by COMT may be responsible for a greater temperament expression in obese individuals. To our knowledge, this is the first study describing the role of affective temperament in the obese population, but more research is needed in this regard.


Assuntos
Catecol O-Metiltransferase/genética , Proteínas da Membrana Plasmática de Transporte de Dopamina/genética , Transtornos do Humor/genética , Obesidade/genética , Polimorfismo Genético , Receptores de Dopamina D4/genética , Adulto , Dopamina/genética , Feminino , Predisposição Genética para Doença , Humanos , Masculino , Pessoa de Meia-Idade , Transtornos do Humor/complicações , Obesidade/complicações , Temperamento
4.
Proc Natl Acad Sci U S A ; 116(9): 3853-3862, 2019 02 26.
Artigo em Inglês | MEDLINE | ID: mdl-30755521

RESUMO

The human dopamine (DA) transporter (hDAT) mediates clearance of DA. Genetic variants in hDAT have been associated with DA dysfunction, a complication associated with several brain disorders, including autism spectrum disorder (ASD). Here, we investigated the structural and behavioral bases of an ASD-associated in-frame deletion in hDAT at N336 (∆N336). We uncovered that the deletion promoted a previously unobserved conformation of the intracellular gate of the transporter, likely representing the rate-limiting step of the transport process. It is defined by a "half-open and inward-facing" state (HOIF) of the intracellular gate that is stabilized by a network of interactions conserved phylogenetically, as we demonstrated in hDAT by Rosetta molecular modeling and fine-grained simulations, as well as in its bacterial homolog leucine transporter by electron paramagnetic resonance analysis and X-ray crystallography. The stabilization of the HOIF state is associated both with DA dysfunctions demonstrated in isolated brains of Drosophila melanogaster expressing hDAT ∆N336 and with abnormal behaviors observed at high-time resolution. These flies display increased fear, impaired social interactions, and locomotion traits we associate with DA dysfunction and the HOIF state. Together, our results describe how a genetic variation causes DA dysfunction and abnormal behaviors by stabilizing a HOIF state of the transporter.


Assuntos
Transtorno do Espectro Autista/genética , Proteínas da Membrana Plasmática de Transporte de Dopamina/genética , Dopamina/genética , Locomoção/genética , Animais , Animais Geneticamente Modificados , Transtorno do Espectro Autista/fisiopatologia , Cristalografia por Raios X , Dopamina/metabolismo , Proteínas da Membrana Plasmática de Transporte de Dopamina/química , Drosophila melanogaster/genética , Drosophila melanogaster/fisiologia , Espectroscopia de Ressonância de Spin Eletrônica , Medo/fisiologia , Humanos , Relações Interpessoais , Locomoção/fisiologia , Modelos Moleculares , Mutação , Deleção de Sequência/genética
5.
Artigo em Inglês | MEDLINE | ID: mdl-30774584

RESUMO

Dopaminergic and serotonergic neurons modulate and control processes ranging from reward signaling to regulation of motor outputs. Further, dysfunction of these neurons is involved in both degenerative and psychiatric disorders. Elucidating the roles of these neurons has been greatly facilitated by bacterial artificial chromosome (BAC) transgenic mouse lines expressing channelrhodopsin to readily enable cell-type specific activation. However, corresponding lines to silence these monoaminergic neurons have been lacking. We have generated two BAC transgenic mouse lines expressing the outward proton pump, enhanced ArchT3.0 (eArchT3.0), and GFP under control of the regulatory elements of either the dopamine transporter (DAT; Jax# 031663) or the tryptophan hydroxylase 2 (TPH2; Jax# 031662) gene locus. We demonstrate highly faithful and specific expression of these lines in dopaminergic and serotonergic neurons respectively. Additionally we validate effective and sensitive eArchT3.0-mediated silencing of these neurons using slice electrophysiology as well as with a well-established behavioral assay. These new transgenic tools will help expedite the study of dopaminergic and serotonergic system function in normal behavior and disease.


Assuntos
Neurônios Dopaminérgicos/fisiologia , Optogenética , Neurônios Serotoninérgicos/fisiologia , Potenciais de Ação/genética , Animais , Encéfalo/citologia , Channelrhodopsins/genética , Channelrhodopsins/metabolismo , Proteínas da Membrana Plasmática de Transporte de Dopamina/genética , Proteínas da Membrana Plasmática de Transporte de Dopamina/metabolismo , Estimulação Elétrica , Vetores Genéticos/genética , Vetores Genéticos/metabolismo , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Células HEK293 , Humanos , Técnicas In Vitro , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Técnicas de Patch-Clamp , RNA Mensageiro/metabolismo , Transfecção , Triptofano Hidroxilase/genética , Triptofano Hidroxilase/metabolismo , Tirosina 3-Mono-Oxigenase/metabolismo
6.
J Biol Chem ; 294(14): 5632-5642, 2019 04 05.
Artigo em Inglês | MEDLINE | ID: mdl-30705091

RESUMO

The human dopamine transporter (hDAT) regulates the level of the neurotransmitter dopamine (DA) in the synaptic cleft and recycles DA for storage in the presynaptic vesicular pool. Many neurotransmitter transporters exist as oligomers, but the physiological role of oligomerization remains unclear; for example, it has been speculated to be a prerequisite for amphetamine-induced release and protein trafficking. Previous studies point to an oligomeric quaternary structure of hDAT; however, the exact stoichiometry and the fraction of co-existing oligomeric states are not known. Here, we used single-molecule brightness analysis to quantify the degree of oligomerization of heterologously expressed hDAT fused to monomeric GFP (mGFP-hDAT) in Chinese hamster ovary (CHO) cells. We observed that monomers and dimers of mGFP-hDAT co-exist and that higher-order molecular complexes of mGFP-hDAT are absent at the plasma membrane. The mGFP-hDAT dimers were stable over several minutes, and the fraction of dimers was independent of the mGFP-hDAT surface density. Furthermore, neither oxidation nor depletion of cholesterol had any effect on the fraction of dimers. Unlike for the human serotonin transporter (hSERT), in which direct binding of phosphatidylinositol 4,5-bisphosphate (PIP2) stabilized the oligomers, the stability of mGFP-hDAT dimers was PIP2 independent.


Assuntos
Membrana Celular/metabolismo , Proteínas da Membrana Plasmática de Transporte de Dopamina/metabolismo , Fosfatidilinositol 4,5-Difosfato/metabolismo , Multimerização Proteica , Animais , Células CHO , Membrana Celular/genética , Colesterol/genética , Colesterol/metabolismo , Cricetulus , Proteínas da Membrana Plasmática de Transporte de Dopamina/genética , Humanos , Fosfatidilinositol 4,5-Difosfato/genética
7.
Psychopharmacology (Berl) ; 236(4): 1349-1365, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30539268

RESUMO

RATIONALE: Dopamine levels are controlled in part by transport across the cell membrane by the dopamine transporter (DAT), and recent evidence showed that a polymorphism in the gene encoding DAT is associated with alcoholism. However, research in animal models using DAT knockout mice has yielded conflicting results. OBJECTIVES: The present study was planned to evaluate the effects of DAT knockdown in the nucleus accumbens (Nacc) on voluntary ethanol consumption and preference in male and female C57BL/6J mice. METHODS: For this purpose, animals were stereotaxically injected with DAT siRNA-expressing lentiviral vectors in the Nacc, and using a voluntary, continuous access two-bottle choice model of alcohol, we investigated the importance of accumbal DAT expression in voluntary alcohol intake and preference. We also investigated the effects of DAT knockdown on saccharin and quinine consumption and ethanol metabolism. RESULTS: We show that females consumed more alcohol than males. Interestingly, DAT knockdown in the Nacc significantly decreased alcohol intake and preference in both groups, but no significant sex by group interaction was observed. Also, DAT knockdown did not alter total fluid consumption, saccharin or quinine consumption, or blood ethanol concentrations. Using Pearson correlation, results indicated a strong positive relationship between DAT mRNA expression and ethanol consumption and preference. CONCLUSIONS: Taken together, these data provide further evidence that DAT plays an important role in controlling ethanol intake and that accumbal DAT contributes in the modulation of the reinforcing effects of ethanol. Overall, the results suggest that DAT inhibitors may be valuable in the pharmacotherapy of alcoholism.


Assuntos
Consumo de Bebidas Alcoólicas/metabolismo , Proteínas da Membrana Plasmática de Transporte de Dopamina/deficiência , Etanol/administração & dosagem , Núcleo Accumbens/metabolismo , Caracteres Sexuais , Consumo de Bebidas Alcoólicas/psicologia , Animais , Dopamina/metabolismo , Proteínas da Membrana Plasmática de Transporte de Dopamina/antagonistas & inibidores , Proteínas da Membrana Plasmática de Transporte de Dopamina/genética , Feminino , Células HEK293 , Humanos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Núcleo Accumbens/efeitos dos fármacos , RNA Interferente Pequeno/farmacologia , Reforço (Psicologia) , Sacarina/administração & dosagem
8.
Ecotoxicol Environ Saf ; 170: 227-237, 2019 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-30529917

RESUMO

Broad applications and exposure to the fungicide maneb can lead to toxicity in non-target organisms. Maneb is also associated with neurogenerative diseases such as Parkinson's disease (PD). The objectives of this study were to determine the acute toxicity of maneb to zebrafish by measuring mitochondrial bioenergetics, locomotor activity, and the expression of genes related to the oxidative damage response, as well as those related to dopamine signaling due to its association with PD. Zebrafish embryos at 6 h post-fertilization (hpf) were exposed to either solvent control (0.1% DMSO, v/v), or one dose of 0.1, 0.5, 1.0 and 10.0 µM maneb for 96 h. Maneb was moderately toxic to zebrafish embryos, and had a 96-h LC50 value of 4.29 µM (~ 1.14 mg/L). Maneb induced a dose-dependent increase in mortality, decreased hatching rate, and increased notochord deformity rate at both 1.0 and 10.0 µM after 72 and 96 h. Total body length was also significantly reduced with 1.0 µM maneb. A 50-60% decrease in mean basal oxygen consumption rate was also observed in embryos following a 24 hpf exposure to 10.0 µM maneb but oligomycin-induced ATP production and FCCP-induced maximum respiration remained unaffected. No change was detected in the expression levels of genes associated with oxidative stress (sod1 and sod2), nor those related to dopamine synthesis (th1), dopamine transporter (dat), dopamine receptors (drd1, drd2a, drd3, and drd4b). Thus, modifying the expression of these transcripts may not be a mechanism for maneb-induced developmental toxicity in zebrafish. To assess the potential for neurotoxicity, a dark photokinesis assay was conducted in larvae following 7 d exposure to 0.1, 0.5 and 1.0 µM maneb. Larvae exposed to 0.5 and 1.0 µM maneb showed signs related to hypoactivity, and this reduced activity is hypothesized to be associated with notochord defects as this deformity was prevalent at higher concentrations of maneb. Overall, these data demonstrate that maneb negatively affects embryonic development (i.e. notochord development), affects basal oxygen consumption rates of embryos, and induces hypoactivity in larval fish. This study improves understanding regarding the developmental neurotoxicity of the fungicide maneb to zebrafish.


Assuntos
Embrião não Mamífero/efeitos dos fármacos , Larva/efeitos dos fármacos , Maneb/toxicidade , Mitocôndrias/efeitos dos fármacos , Notocorda/efeitos dos fármacos , Peixe-Zebra/embriologia , Animais , Proteínas da Membrana Plasmática de Transporte de Dopamina/genética , Proteínas da Membrana Plasmática de Transporte de Dopamina/metabolismo , Relação Dose-Resposta a Droga , Embrião não Mamífero/patologia , Desenvolvimento Embrionário/efeitos dos fármacos , Metabolismo Energético/efeitos dos fármacos , Feminino , Expressão Gênica , Locomoção/efeitos dos fármacos , Masculino , Mitocôndrias/patologia , Notocorda/patologia , Estresse Oxidativo/efeitos dos fármacos , Consumo de Oxigênio , Praguicidas/toxicidade , Receptores Dopaminérgicos/genética , Receptores Dopaminérgicos/metabolismo , Superóxido Dismutase/genética , Superóxido Dismutase/metabolismo , Superóxido Dismutase-1/genética , Superóxido Dismutase-1/metabolismo , Peixe-Zebra/metabolismo
9.
J Biol Chem ; 294(10): 3419-3431, 2019 03 08.
Artigo em Inglês | MEDLINE | ID: mdl-30587577

RESUMO

Several protein kinases, including protein kinase C, Ca2+/calmodulin-dependent protein kinase II, and extracellular signal-regulated kinase, play key roles in the regulation of dopamine transporter (DAT) functions. These functions include surface expression, internalization, and forward and reverse transport, with phosphorylation sites for these kinases being linked to distinct regions of the DAT N terminus. Protein phosphatases (PPs) also regulate DAT activity, but the specific residues associated with their activities have not yet been elucidated. In this study, using co-immunoprecipitation followed by MS and immunoblotting analyses, we demonstrate the association of DAT with PP1 and PP2A in the mouse brain and heterologous cell systems. By applying MS in conjunction with a metabolic labeling method, we defined a PP1/2A-sensitive phosphorylation site at Thr-48 in human DAT, a residue that has not been previously reported to be involved in DAT phosphorylation. Site-directed mutagenesis of Thr-48 to Ala (T48A) to prevent phosphorylation enhanced dopamine transport kinetics, supporting a role for this residue in regulating DAT activity. Moreover, T48A-DAT displayed increased palmitoylation, suggesting that phosphorylation/dephosphorylation at this site has an additional regulatory role and reinforcing a previously reported reciprocal relationship between C-terminal palmitoylation and N-terminal phosphorylation.


Assuntos
Encéfalo/metabolismo , Proteínas da Membrana Plasmática de Transporte de Dopamina/metabolismo , Dopamina/metabolismo , Proteína Fosfatase 1/metabolismo , Proteína Fosfatase 2/metabolismo , Animais , Transporte Biológico Ativo/fisiologia , Dopamina/genética , Proteínas da Membrana Plasmática de Transporte de Dopamina/genética , Humanos , Lipoilação/genética , Camundongos , Camundongos Knockout , Fosforilação , Proteína Fosfatase 1/genética , Proteína Fosfatase 2/genética , Treonina/genética , Treonina/metabolismo
10.
J Mol Neurosci ; 65(4): 527-535, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-30083784

RESUMO

The purpose of this study was to explore the key mechanism involved in the pathogenesis of Parkinson's disease (PD) based on microarray analysis. The expression profile data of GSE7621, which contained 9 substantia nigra tissues isolated from normals and 16 substantia nigra tissues isolated from PD patients, was obtained from Gene Expression Omnibus. The differentially expressed genes (DEGs) were screened, followed by functional enrichment analysis and protein-protein interaction (PPI) network construction. After the miRNAs regulating the DEGs were predicted, the miRNA-DEG regulatory network was then constructed. Besides, the 6-hydroxydopamine rat model of PD was established and the expression of key DEGs and miRNA was detected. A total of 388 DEGs were identified, including 218 upregulated genes and 170 downregulated ones. Tyrosine hydroxylase (TH) and solute carrier family 6 member 3 (SLC6A3) were significantly related to the functional terms of catecholamine biosynthetic process and dopamine biosynthetic process. TH and SLC6A3 were hub nodes in the PPI network. EBF3 could be targeted by miR-218. Moreover, TH and SLC6A3 were found downregulated in the 6-OHDA rat model of PD, while miR-218 was markedly upregulated. Our results reveal that SLC6A3, TH, and EBF3 targeted by miR-218 could be involved in PD. These molecules might provide a new insight into the development of therapeutic strategies for PD.


Assuntos
Redes Reguladoras de Genes , MicroRNAs/genética , Doença de Parkinson/genética , Mapas de Interação de Proteínas , Animais , Proteínas da Membrana Plasmática de Transporte de Dopamina/genética , Proteínas da Membrana Plasmática de Transporte de Dopamina/metabolismo , Humanos , Masculino , Ratos , Ratos Wistar , Substância Negra/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Tirosina 3-Mono-Oxigenase/genética , Tirosina 3-Mono-Oxigenase/metabolismo
11.
J Biomed Sci ; 25(1): 61, 2018 Aug 07.
Artigo em Inglês | MEDLINE | ID: mdl-30086746

RESUMO

BACKGROUND: Studies at the molecular level aim to integrate genetic and neurobiological data to provide an increasingly detailed understanding of phenotypes related to the ability in time perception. MAIN TEXT: This study suggests that the polymorphisms genetic SLC6A4 5-HTTLPR, 5HTR2A T102C, DRD2/ANKK1-Taq1A, SLC6A3 3'-UTR VNTR, COMT Val158Met, CLOCK genes and GABRB2 A/C as modification factor at neurochemical levels associated with several neurofunctional aspects, modifying the circadian rhythm and built-in cognitive functions in the timing. We conducted a literature review with 102 studies that met inclusion criteria to synthesize findings on genetic polymorphisms and their influence on the timing. CONCLUSION: The findings suggest an association of genetic polymorphisms on behavioral aspects related in timing. However, order to confirm the paradigm of association in the timing as a function of the molecular level, still need to be addressed future research.


Assuntos
Ritmo Circadiano/genética , Cognição/fisiologia , Predisposição Genética para Doença , Percepção do Tempo/fisiologia , Adulto , Ritmo Circadiano/fisiologia , Proteínas da Membrana Plasmática de Transporte de Dopamina/genética , Feminino , Estudos de Associação Genética , Genótipo , Humanos , Masculino , Polimorfismo de Nucleotídeo Único/genética , Proteínas Serina-Treonina Quinases/genética , Receptores de Dopamina D2/genética , Proteínas da Membrana Plasmática de Transporte de Serotonina/genética
12.
Cell Mol Life Sci ; 75(23): 4357-4370, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30043140

RESUMO

Melatonin, a neuro-hormone released by the pineal gland, has multiple effects in the central nervous system including the regulation of dopamine (DA) levels, but how melatonin accomplishes this task is not clear. Here, we show that melatonin MT1 and MT2 receptors co-immunoprecipitate with the DA transporter (DAT) in mouse striatal synaptosomes. Increased DA re-uptake and decreased amphetamine-induced locomotor activity were observed in the striatum of mice with targeted deletion of MT1 or MT2 receptors. In vitro experiments confirmed the interactions and recapitulated the inhibitory effect of melatonin receptors on DA re-uptake. Melatonin receptors retained DAT in the endoplasmic reticulum in its immature non-glycosylated form. In conclusion, we reveal one of the first molecular complexes between G protein-coupled receptors (MT1 and MT2) and transporters (DAT) in which melatonin receptors regulate the availability of DAT at the plasma membrane, thus limiting the striatal DA re-uptake capacity in mice.


Assuntos
Membrana Celular/metabolismo , Proteínas da Membrana Plasmática de Transporte de Dopamina/metabolismo , Dopamina/metabolismo , Receptor MT1 de Melatonina/metabolismo , Receptor MT2 de Melatonina/metabolismo , Animais , Corpo Estriado/metabolismo , Proteínas da Membrana Plasmática de Transporte de Dopamina/genética , Células HEK293 , Humanos , Masculino , Camundongos Endogâmicos C57BL , Camundongos Knockout , Atividade Motora/genética , Ligação Proteica , Receptor MT1 de Melatonina/genética , Receptor MT2 de Melatonina/genética , Sinaptossomos/metabolismo
13.
Yonsei Med J ; 59(6): 787-792, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-29978616

RESUMO

PURPOSE: The present study investigated associations between dopamine transporter (DAT) availability and α-synuclein levels in cerebrospinal fluid, as well as synuclein gene (SNCA) transcripts, and the effect of single nucleotide polymorphism of SNCA on DAT availability in healthy subjects. MATERIALS AND METHODS: The study population comprised healthy controls who underwent ¹²³I-FP-CIT single-photon emission computed tomography screening. Five SNCA probes were used to target the boundaries of exon 3 and exon 4 (SNCA-E3E4), transcripts with a long 3'UTR region (SNCA-3UTR-1, SNCA-3UTR-2), transcripts that skip exon 5 (SNCA-E4E6), and the rare short transcript isoforms that comprise exons 1-4 (SNCA-007). RESULTS: In total, 123 healthy subjects (male 75, female 48) were included in this study. DAT availability in the caudate nucleus (p=0.0661) and putamen (p=0.0739) tended to differ according to rs3910105 genotype. In post-hoc analysis, DAT availability in the putamen was lower in subjects of TT genotype than those of CC/CT (p=0.0317). DAT availability in the caudate nucleus also showed a trend similar to that in the putamen (p=0.0597). Subjects of CT genotype with rs3910105 showed negative correlations with DAT availability in the putamen with SNCA-E3E4 (p=0.037, rho=-0.277), and SNCA-E4E6 (p=0.042, rho=-0.270), but not those of CC/TT genotypes. CONCLUSION: This is the first study to investigate the association of rs3910105 in SNCA with DAT availability. rs3910105 had an effect on DAT availability, and the correlation between DAT availability and SNCA transcripts were significant in CT genotypes of rs3910105.


Assuntos
Líquido Cefalorraquidiano/metabolismo , Proteínas da Membrana Plasmática de Transporte de Dopamina/metabolismo , Voluntários Saudáveis , Tomografia Computadorizada de Emissão de Fóton Único , alfa-Sinucleína/genética , Adulto , Proteínas da Membrana Plasmática de Transporte de Dopamina/genética , Feminino , Expressão Gênica , Genótipo , Humanos , Masculino , Pessoa de Meia-Idade , Polimorfismo de Nucleotídeo Único/genética , Tomografia Computadorizada de Emissão de Fóton Único/métodos , Tropanos , alfa-Sinucleína/metabolismo
14.
Psychiatr Danub ; 30(1): 47-56, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29546858

RESUMO

BACKGROUND: the objective of this study was to examine the associations between Cloninger temperament and character dimensions with the DAT1 VNTR and COMT Val158Met polymorphisms. SUBJECTS AND METHODS: The study was conducted on 101 subjects, consisting of students of the Police College in Zagreb and staff of the Sestre Milosrdnice University Hospital in Zagreb. The Cloninger Temperament and Character Inventory (TCI) was used to test personality traits. RESULTS: A main effect of the DAT1 VNTR polymorphism was found on the subscale self-directedness - SD2 (F=5.18, df=1, p<0.05), where a higher result was detected in carriers of the 9/9 genotype (M=7.33, SD=0.51) than those carrying the 10-repeat allele (M=6.02, SD=1.36). Also for the COMT Val158Met polymorphism, main effects were found on the subscales: NS3 (novelty seeking) (F=5.18, df=1, p<0.05), where a higher result was found in carriers of the Val allele (M=5.03, SD=2.22) than in carriers of the Met/Met genotype (MD=4.76, SD=2.37), SD3 (self-directedness) (F=5.18, df=1, p<0.05) where a higher result was found in carriers of the Val/Val genotype (M=4.50, SD=0.78) than in those carrying the Met allele (M=3.80, SD=1.31); C3 (cooperativeness) (F=5.18, df=1, p<0.05), where a high result was found in carriers of the Val allele (M=5.68, SD=1.25) than those carrying the Met/Met genotype (M=5.08, SD=1.11); and ST3 (self-transcendence) (F=5.18, df=1, p<0.05), where a higher result was found in carriers of the Met/Met genotype (M=3.46, SD=2.37) than carriers of the Val allele (M=2.69, SD=1.84). Two significant interactions were detected, on the subscale NS3 (novelty seeking) (F=5.18, df=1, p<0.05), and on the subscale C2 (cooperativeness) (F=5.18, df=1, p<0.05). CONCLUSIONS: Cloninger's (1987) hypothesis about negative relationship between novelty seeking and dopamine was confirmed on allele level, because higher novelty seeking was found in Val allele carriers comparing to Met/Met genotype carriers.


Assuntos
Catecol O-Metiltransferase/genética , Caráter , Proteínas da Membrana Plasmática de Transporte de Dopamina/genética , Metionina/genética , Polimorfismo Genético/genética , Temperamento , Valina/genética , Adulto , Alelos , Croácia , Dopamina/fisiologia , Comportamento Exploratório/fisiologia , Feminino , Triagem de Portadores Genéticos , Genótipo , Humanos , Masculino , Repetições Minissatélites/genética , Determinação da Personalidade/estatística & dados numéricos , Psicometria
15.
Neuropharmacology ; 133: 216-223, 2018 05 01.
Artigo em Inglês | MEDLINE | ID: mdl-29407766

RESUMO

Patients afflicted with bipolar disorder demonstrate significant impairments in recognition and episodic memory during acute depressive and manic episodes. These impairments and the related pathophysiology may result from over-activation of the brain dopamine (DA) system. In order to model overactive DA transmission in a well-established novel object recognition (NOR) memory test, we used DA transporter knockdown (DAT-KD) mice, which exhibit reduced DAT expression and display hyper-dopaminergic phenotypes. DAT-KD mice exhibited impaired NOR memory compared to wild-type (WT) mice. This impairment was prevented by administration of FAUC365, a DA D3 receptor (D3R) selective antagonist, prior to object learning. Similarly, D3R knockout (KO)/DAT-KD double mutant mice displayed performance in the NOR test that was comparable to WT mice, suggesting that deficiencies in NOR performance in DAT-KD mice can be compensated by diminishing D3R signaling. GBR12909, a DAT blocker, also impaired NOR performance in WT mice, but not in D3R KO mice. Impaired NOR performance in GBR12909-treated WT mice was also prevented by pretreatment with FAUC365. Together, these findings indicate that reduced DAT activity can impair recognition memory in the NOR test, and D3R appears to be necessary to mediate this effect.


Assuntos
Dopamina/metabolismo , Transtornos da Memória/tratamento farmacológico , Transtornos da Memória/genética , Receptores de Dopamina D3/deficiência , Recognição (Psicologia)/efeitos dos fármacos , Análise de Variância , Animais , Modelos Animais de Doenças , Antagonistas de Dopamina/uso terapêutico , Proteínas da Membrana Plasmática de Transporte de Dopamina/deficiência , Proteínas da Membrana Plasmática de Transporte de Dopamina/genética , Indóis/uso terapêutico , Aprendizagem em Labirinto/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Piperazinas/uso terapêutico , Piperidinas/uso terapêutico , Receptores de Dopamina D3/genética , Tiofenos/uso terapêutico
16.
Genes Brain Behav ; 17(4): e12463, 2018 04.
Artigo em Inglês | MEDLINE | ID: mdl-29406596

RESUMO

Attention deficit hyperactivity disorder (ADHD) is a neuropsychiatric disorder characterized by a developmentally inappropriate, pervasive and persistent pattern of severe inattention, hyperactivity and impulsivity. Despite onset in early childhood, ADHD may continue into adulthood with substantial impairment in social, academic and occupational functioning. A new animal model of this disorder was developed in rats with genetic deletion of the dopamine transporter (DAT) gene (dopamine transporter knockout rats; DAT-KO rats). We analyzed the behavior of DAT-KO rats for a deeper phenotypical characterization of this model. We first tested rats of the 3 genotypes at different ages (preadolescent, adolescent and adult), in a novelty-seeking test using a black/white box (Experiment 1). After that, we tested adult rats in a novelty-preference test using a 3-chamber apparatus with different shapes (Experiment 2). Experiment 1: as evidenced by analysis of time spent in the novel environment, adult DAT heterozygous (DAT-HET) rats show an increased curiosity-driven exploration compared with wild-type (WT) controls while DAT-KO rats did not recognize novelty. The locomotor activity data show a minimal difference between genotypes at adolescent age while the preadolescent and adult DAT-KO rats have significantly increased activity rate compared with WT and DAT-HET subjects. Experiment 2: in this case, due to more clearly evident spatial differences, time spent in novel environment was not significantly different among genotypes. During first 10 minutes, DAT-KO rats showed a decreased hyperactivity, apparently related to curiosity and attention to the new environments. In conclusion, DAT-KO rats may show some inattention while more novelty-seeking traits appear in DAT-HET rats.


Assuntos
Proteínas da Membrana Plasmática de Transporte de Dopamina/genética , Comportamento Exploratório/fisiologia , Fatores Etários , Animais , Transtorno do Deficit de Atenção com Hiperatividade/genética , Cognição/fisiologia , Modelos Animais de Doenças , Emoções/fisiologia , Feminino , Técnicas de Inativação de Genes , Comportamento Impulsivo/fisiologia , Masculino , Atividade Motora/genética , Ratos , Ratos Wistar
17.
Genes Brain Behav ; 17(4): e12460, 2018 04.
Artigo em Inglês | MEDLINE | ID: mdl-29377542

RESUMO

Dopamine (DA) plays a significant role in cognition, motor function and social behavior. The objectives of this study were to (1) quantify the temporal expression of transcripts (DA receptors, transporters and tyrosine hydroxylase) associated with DA signaling during early stages of zebrafish development and (2) determine their expression profiles following treatment with a D2 receptor antagonist domperidone (DMP). We also assessed locomotor behavior following treatment with DMP using alternating periods of light and dark (ie, dark photokinesis), as DA plays a key role in behavior. Relative expression levels of transcripts that were investigated and related to the DA system were detected after the first 24 hours postfertilization (hpf). Some DA receptor transcripts (eg, drd4c) increased in abundance earlier in the embryo compared with other receptors (eg, drd3), suggesting that DA receptor paralogs may have unique roles in development. Treatment of larvae with DMP resulted in the upregulation of DA receptor transcripts (ie, drd1, drd7, drd4b, drd4c) and DA transporter 1 (ie, slc6a3), and it is hypothesized that upregulation of genes related to the DA system is a compensatory neurophysiological response to DA receptor antagonism. Larval activity during dark photokinesis (measured by distance traveled) was also elevated by DMP. We hypothesize that behavioral responses observed with DMP may be related to the regulation of deep brain photoreception in zebrafish (Danio rerio) (ZF) larvae by DA.


Assuntos
Domperidona/farmacologia , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Receptores Dopaminérgicos/efeitos dos fármacos , Animais , Estimulantes do Sistema Nervoso Central/farmacologia , Domperidona/metabolismo , Dopamina/metabolismo , Antagonistas de Dopamina/farmacologia , Proteínas da Membrana Plasmática de Transporte de Dopamina/genética , Larva , Locomoção/efeitos dos fármacos , Receptores Dopaminérgicos/genética , Receptores de Dopamina D2/genética , Ativação Transcricional , Transcriptoma/genética , Tirosina 3-Mono-Oxigenase/genética , Regulação para Cima , Peixe-Zebra/embriologia , Peixe-Zebra/genética , Peixe-Zebra/metabolismo
18.
Neuropharmacology ; 133: 171-180, 2018 05 01.
Artigo em Inglês | MEDLINE | ID: mdl-29378213

RESUMO

This study aimed to address the mechanisms and reinforcing effects of three synthetic cathinone analogs of MDMA commonly reported in "Ecstasy" formulations: methylone, butylone, and pentylone. Whole-cell patch clamp techniques were used to assess the mechanism of each compound at the dopamine and serotonin transporters. Separate groups of rats were trained to discriminate methamphetamine, DOM, or MDMA from vehicle. Substitution studies were performed in each group and antagonism studies with SCH23390 were performed against each compound that produced substitution. Self-administration of each compound was evaluated under a progressive ratio schedule of reinforcement. Each compound produced an inward current at the serotonin transporter, but little or no current at the dopamine transporter. Each of the test compounds substituted fully for the discriminative stimulus effects of methamphetamine, methylone and butylone substituted partially for DOM and fully for MDMA, whereas pentylone failed to substitute for DOM and substituted only partially for MDMA. SCH23390 fully and dose-dependently attenuated methamphetamine-appropriate responding produced by each test compound, but was least potent against pentylone. MDMA-appropriate responding was minimally affected by SCH23390. Each test compound was robustly self-administered with pentylone producing the greatest self-administration at the doses tested. Given the prevalence of synthetic cathinones in "Ecstasy" formulations, these data indicate that adulterated "Ecstasy" formulations may drive more compulsive drug use than those containing only MDMA.


Assuntos
Alucinógenos/farmacologia , N-Metil-3,4-Metilenodioxianfetamina/farmacologia , Reforço (Psicologia) , Transtornos Relacionados ao Uso de Substâncias/fisiopatologia , Animais , Benzazepinas/farmacologia , Estimulantes do Sistema Nervoso Central/farmacologia , Condicionamento Operante/efeitos dos fármacos , Modelos Animais de Doenças , Antagonistas de Dopamina/farmacologia , Proteínas da Membrana Plasmática de Transporte de Dopamina/genética , Proteínas da Membrana Plasmática de Transporte de Dopamina/metabolismo , Combinação de Medicamentos , Células HEK293 , Humanos , Masculino , Metanfetamina/análogos & derivados , Metanfetamina/farmacologia , N-Metil-3,4-Metilenodioxianfetamina/análogos & derivados , Técnicas de Patch-Clamp , Ratos , Ratos Sprague-Dawley , Autoadministração , Serotonina/farmacologia , Transtornos Relacionados ao Uso de Substâncias/etiologia
19.
Int J Mol Med ; 41(1): 421-429, 2018 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-29115391

RESUMO

The herbicide simazine is widely used in agricultural and non-agricultural fields. Studies have shown that simazine inhibits the proliferation of dopaminergic cells and affects the developmental differentiation of dopamine neurons. However, little is known about the effects of simazine on dopaminergic metabolism. Therefore, the present study examined the effects of simazine on Sprague­Dawley (SD) rats from weaning to puberty (40 days exposure). Simazine was administered orally to SD rats at doses of 0, 12.5, 50 and 200 mg/kg body weight. The contents of dopamine (DA), levodopa, dihydroxy-phenyl-acetic acid and homovanillic acid in the striatum were then examined by high-performance liquid chromatography with a fluorescence detector. Quantitative polymerase chain reaction and western blotting were used to analyze the mRNA and protein expression of aromatic amino acid decarboxylase (AADC), tyrosine hydroxylase, orphan nuclear hormone (Nurr1), dopamine transporter (DAT), vesicular monoamine transporter 2 (VMAT2), monoamine oxidase (MAO) and catechol-O-methyltransferase (COMT). The results indicated that simazine influenced the synthesis, transport and metabolism of DA and led to a reduction of DA levels in the striatum. One potential underlying mechanism is decreased levels of Nurr1, DAT and VMAT2 impacting upon the transport of DA; another is the decreased level of AADC and increased levels of MAO and COMT impacting upon the synthesis and metabolism of DA. These factors may eventually lead to neurological disorders of the dopaminergic system.


Assuntos
Dopamina/metabolismo , Neurônios Dopaminérgicos/efeitos dos fármacos , Simazina/toxicidade , Animais , Descarboxilases de Aminoácido-L-Aromático/genética , Catecol O-Metiltransferase/genética , Proliferação de Células/efeitos dos fármacos , Proliferação de Células/genética , Dopamina/biossíntese , Dopamina/genética , Proteínas da Membrana Plasmática de Transporte de Dopamina/genética , Neurônios Dopaminérgicos/metabolismo , Neurônios Dopaminérgicos/patologia , Regulação da Expressão Gênica/efeitos dos fármacos , Monoaminoxidase/genética , Membro 2 do Grupo A da Subfamília 4 de Receptores Nucleares/genética , Ratos , Tirosina 3-Mono-Oxigenase/genética , Proteínas Vesiculares de Transporte de Monoamina/genética
20.
Arch Physiol Biochem ; 124(4): 330-334, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-29145734

RESUMO

Obesity is becoming an epidemic in Latin American countries. Genetic analyses of endophenotypes for obesity, such as body mass index (BMI), are quite useful for research. In this study, we analysed two functional polymorphisms in the dopamine receptor 4 (DRD4) and dopamine transporter (SLC6A3) genes. A sample of 232 Colombian young subjects were recruited and evaluated for BMI. Two functional polymorphisms in the DRD4 and SLC6A3 and genes were genotyped by PCR and electrophoresis. A significant association was found between BMI and the polymorphisms in DRD4 and SLC6A3 genes. DRD4 4/4 genotype was associated with a lower mean BMI and SLC6A3 10/10 genotype was associated with a higher mean BMI. Our work provides additional novel findings about the association of dopaminergic genes with BMI in healthy young adults. In addition, our study is one the first analyses of candidate genes for BMI in Latin American samples.


Assuntos
Proteínas da Membrana Plasmática de Transporte de Dopamina/genética , Predisposição Genética para Doença , Sobrepeso/genética , Polimorfismo Genético , Receptores de Dopamina D4/genética , Adolescente , Adulto , Alelos , Animais , Índice de Massa Corporal , Estudos de Casos e Controles , Colômbia , Proteínas da Membrana Plasmática de Transporte de Dopamina/metabolismo , Feminino , Frequência do Gene , Estudos de Associação Genética , Humanos , Masculino , Sobrepeso/sangue , Sobrepeso/metabolismo , Obesidade Pediátrica/sangue , Obesidade Pediátrica/genética , Obesidade Pediátrica/metabolismo , Receptores de Dopamina D4/metabolismo , Estudantes de Medicina , Adulto Jovem
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