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1.
Biochem Pharmacol ; 175: 113870, 2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-32088264

RESUMO

The 5-HT1A and 5-HT1B serotonin receptors are abundantly expressed in the CNS and constitute validated as well as putative drug targets in a variety of psychiatric and cognitive disorders, alcoholism/addiction, pain and migraine. In the present study we have characterized the functional properties of human 5-HT1A and 5-HT1B stably co-expressed with the human G-protein-activated inwardly rectifying K+ channel 2 (GIRK2) in HEK293 cells in the fluorescence-based FLIPR® Membrane Potential Blue (FMP) assay. Serotonin and other agonists induced robust decreases in fluorescence levels in the 5-HT1A/GIRK2- and 5-HT1B/GIRK2-HEK293 cells in a concentration-dependent manner in the assay, and these responses could be inhibited by selective 5-HT1A/5-HT1B antagonists and by the Gαi/o-protein inhibitor pertussis toxin (PTX). Five additional stable HEK293 cell lines co-expressing 5-HT1A or 5-HT1B with GIRK2 and one of the PTX-insensitive Gαi/o-subunit mutants Gαi1C351I, Gαi2C352I and Gαo1C351I were constructed, and 5-HT1A/5-HT1B-mediated responses through these specific Gαi/o-subunits were measured in these cells pretreated with PTX in the FMP assay. The functional properties of 16 reference 5-HT1 agonists were characterized at the seven cell lines, which constitutes the most detailed pharmacological profiling and comparison of 5-HT1A and 5-HT1B receptor signaling in the same assay published to date. We propose that this approach to assay 5-HT1-mediated signaling through endogenous Gαi/o-proteins in HEK293 cells or through specific Gαi/o-subunits in a fairly high-throughput manner holds some advantages to other functional assays for Gαi/o-coupled receptors. The assay will facilitate detailed profiling of the Gαi/o- and Gßγ-mediated signaling of 5-HT1A and 5-HT1B at the molecular level, and it could also be used to identify novel modulators for the receptors.


Assuntos
Canais de Potássio Corretores do Fluxo de Internalização Acoplados a Proteínas G/metabolismo , Potenciais da Membrana , Receptor 5-HT1A de Serotonina/metabolismo , Receptor 5-HT1B de Serotonina/metabolismo , Bioensaio , Relação Dose-Resposta a Droga , Fluorescência , Canais de Potássio Corretores do Fluxo de Internalização Acoplados a Proteínas G/genética , Células HEK293 , Humanos , Ligantes , Potenciais da Membrana/efeitos dos fármacos , Bloqueadores dos Canais de Potássio/farmacologia , Receptor 5-HT1A de Serotonina/genética , Receptor 5-HT1B de Serotonina/genética , Agonistas do Receptor 5-HT1 de Serotonina/farmacologia , Antagonistas do Receptor 5-HT1 de Serotonina/farmacologia , Transdução de Sinais , Transfecção
2.
Pharmacol Rep ; 72(2): 427-434, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32002826

RESUMO

BACKGROUND: In our previous work, spiroxatrine was taken as reference compound to develop selective NOP ligands. Therefore, several triazaspirodecanone derivatives were synthesized. Here, we verify their selectivity towards other 5-HT1 receptor subtypes and with respect to α2-AR (Adrenergic Receptors). METHODS: Binding affinities were determined on cells expressing human cloned receptors for 5-HT1A/B/D and α2A/B/C subtypes. The Ki values were determined for those with at least 50% radioligand inhibition. RESULTS: All our derivatives show a moderate affinity for α2 subtypes, spanning from 5 to 7.5 pKi values. Moreover, they show affinity values in a µM-nM range at the 5-HT1A receptor, while they are practically inactive at 5-HT1B and 5-HT1D subtypes. Compound 11, the best of the series, has a 5-HT1A pKi value of 8.43 similar to spiroxatrine but, notably, it has a 5-HT1A favorable selectivity ratio of 52, 8 and 29, respectively over α2A, α2B and α2C adrenoceptor subtypes. CONCLUSIONS: In this SAR study, a 5-HT1A selective ligand has been identified in which a tetralone moiety replaced the 1,4-benzodioxane of spiroxatrine and the methylene linker to the triazaspirodecanone portion was maintained in position 2.


Assuntos
Dioxanos/farmacologia , Descoberta de Drogas , Receptor 5-HT1A de Serotonina/metabolismo , Compostos de Espiro/farmacologia , Animais , Ligação Competitiva , Células CHO , Cricetulus , Dioxanos/química , Dioxanos/metabolismo , Humanos , Ligantes , Estrutura Molecular , Ligação Proteica , Ensaio Radioligante , Receptor 5-HT1A de Serotonina/genética , Receptores Adrenérgicos alfa 2/genética , Receptores Adrenérgicos alfa 2/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Compostos de Espiro/química , Compostos de Espiro/metabolismo , Relação Estrutura-Atividade
3.
Drug Dev Res ; 81(1): 102-113, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31617956

RESUMO

The severity of symptoms as well as efficacy of antidepressants in major depressive disorder (MDD) is modified by single nucleotide polymorphisms (SNPs) in different genes, which may contribute in an additive or synergistic fashion. We aimed to investigate depression severity in participants with MDD under treatment with antidepressants in relation to the combinatory effect of selected genetic variants combined using a genetic risk score (GRS). The sample included 150 MDD patients on regular AD therapy from the population-based Swiss PsyCoLaus cohort. We investigated 44 SNPs previously associated with antidepressant response by ranking them with regard to their association to the Center for Epidemiologic Studies Short Depression Scale (CES-D) score using random forest. The three top scoring SNPs (rs12248560, rs878567, rs17710780) were subsequently combined into an unweighted GRS, which was included in linear and logistic regression models using the CES-D score, occurrence of a major depressive episode (MDE) during follow-up and regular antidepressant treatment during the 6 months preceding follow-up assessment as outcomes. The GRS was associated with MDE occurrence (p = .02) and ln CES-D score (p = .001). The HTR1A rs878567 variant was associated with ln CES-D after adjustment for demographic and clinical variables [p = .02, lower scores for minor allele (G) carriers]. Additionally, rs12248560 (CYP2C19) CC homozygotes showed a six-fold higher likelihood of regular AD therapy at follow-up compared to minor allele homozygotes [TT; ultrarapid metabolizers (p = .03)]. Our study suggests that the cumulative consideration of pharmacogenetic risk variants more reliably reflects the impact of the genetic background on depression severity than individual SNPs.


Assuntos
Antidepressivos/uso terapêutico , Citocromo P-450 CYP2C19/genética , Transtorno Depressivo Maior/tratamento farmacológico , Polimorfismo de Nucleotídeo Único , Receptor 5-HT1A de Serotonina/genética , Adulto , Idoso , Transtorno Depressivo Maior/genética , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Variantes Farmacogenômicos , Estudos Prospectivos , Índice de Gravidade de Doença , Suíça , Resultado do Tratamento
4.
Rev. int. androl. (Internet) ; 17(4): 138-142, oct.-dic. 2019. tab
Artigo em Inglês | IBECS | ID: ibc-189271

RESUMO

INTRODUCTION AND OBJECTIVES: Lifelong premature ejaculation (LPE) is identified as the inability to delay ejaculation for more than 1min after vaginal penetration occurring on all or almost all sexual experiences together with feelings of frustration of both the patient and his partner with avoidance of sexual intimacy. Recently, a role for (HTR1A)-C (1019) G gene polymorphism in patients with LPE was postulated. MATERIALS AND METHODS: Three hundred and fifty participants were prospectively enrolled in this study. They were recruited from the outpatient clinic of Andrology & STDs Department Cairo University from December 2015 to January 2017. Two hundred and forty-five of them were suffering from lifelong premature ejaculation joined this study, in addition to 105 controls. We instructed the wives of the patients to measure the intra-vaginal ejaculation latency time (IELT) of the first intercourse only using a stopwatch for 1 month. Genotyping was performed at the end of the study. RESULTS: The results showed that the majority of the patients were CG, while; the controls were GG. This difference revealed a statistically significant association (p-value<0.001). A highly significant statistical association was found between the studied gene polymorphisms and the IELT among cases (p-values=0.001). CONCLUSION: The study replicated the potential role of 5HT-1A receptor gene polymorphisms in patients with lifelong premature ejaculation


INTRODUCCIÓN Y OBJETIVOS: La eyaculación precoz permanente (LPE) se identifica como la incapacidad para retrasar la eyaculación más de 1min después de la penetración vaginal, que en todas o casi todas las experiencias sexuales provoca sentimientos de frustración tanto en el paciente como en su pareja y conduce a la abstención de las relaciones sexuales. Recientemente, se ha propuesto que el polimorfismo del gen (HTR1A)-C (1019) G tiene un papel en pacientes con LPE. MATERIALES Y MÉTODOS: Se incluyó a 350 participantes en este estudio. Se los reclutó en la clínica ambulatoria del Departamento de Andrología y ETS de la Universidad del Cairo entre diciembre de 2015 y enero de 2017. Doscientos cuarenta y cinco de ellos con eyaculación precoz permanente se incorporaron a este estudio, además de 105 controles. Instruimos a las esposas de los pacientes para medir el tiempo de latencia de la eyaculación (TLE) intravaginal de la primera relación sexual utilizando solamente un cronómetro durante 1 mes. La genotipificación se realizó al final del estudio. RESULTADOS: Los resultados mostraron que la mayoría de los pacientes fueron CG, mientras los controles fueron GG. Esta diferencia reveló una asociación estadísticamente significativa (valor de p < 0,001). Se encontró una asociación estadística muy significativa entre los polimorfismos de los genes estudiados y el TLE entre casos (valores de p = 0,001). CONCLUSIÓN: El estudio reprodujo el papel potencial de los polimorfismos del gen del receptor de 5-HT1A en pacientes con eyaculación precoz permanente


Assuntos
Humanos , Masculino , Adulto Jovem , Adulto , Polimorfismo Genético , Ejaculação Precoce/genética , Receptor 5-HT1A de Serotonina/genética , Ejaculação Precoce/fisiopatologia , Estudos Prospectivos
5.
Genes (Basel) ; 10(10)2019 10 12.
Artigo em Inglês | MEDLINE | ID: mdl-31614865

RESUMO

This study identified a transcription factor that might bind to the 5' regulatory region of the HTR1A and explored the potential effect on 5-HT1A receptor expression. Based on JASPAR predictions, the binding of the transcription factor was demonstrated using the electrophoretic mobility shift assay (EMSA). Vectors over-expressing the transcription factor were co-transfected into HEK-293 and SK-N-SH cells with the recombinant pGL3 vector, and relative fluorescence intensity was measured to determine regulatory activity. Additionally, the qRT-PCR and Western blot were also used to identify whether the transcription factor modulated the endogenous expression of 5-HT1A receptor. The results suggest that the transcription factor CCAA/T enhancer binding protein beta (CEBPB) likely binds to the -1219 to -1209 bp (ATG+1) region of the HTR1A. Two sequences located in the -722 to -372 bp and -119 to +99 bp were also identified. Although the effect of CEBPB on endogenous 5-HT1A receptor expression was not significant, it exhibited the strong inhibition on the relative fluorescence intensity and the mRNA level of HTR1A. CEBPB inhibited the human HTR1A expression by binding to the sequence -1219 - -1209 bp. This is useful and informative for ascertaining the regulation of 5-HT1A receptor and mental diseases.


Assuntos
Região 5'-Flanqueadora , Proteína beta Intensificadora de Ligação a CCAAT/metabolismo , Receptor 5-HT1A de Serotonina/genética , Regulação da Expressão Gênica , Células HEK293 , Humanos , Receptor 5-HT1A de Serotonina/metabolismo , Sequências Reguladoras de Ácido Nucleico
6.
Nat Commun ; 10(1): 3924, 2019 09 02.
Artigo em Inglês | MEDLINE | ID: mdl-31477731

RESUMO

The serotonergic system and in particular serotonin 1A receptor (5-HT1AR) are implicated in major depressive disorder (MDD). Here we demonstrated that 5-HT1AR is palmitoylated in human and rodent brains, and identified ZDHHC21 as a major palmitoyl acyltransferase, whose depletion reduced palmitoylation and consequently signaling functions of 5-HT1AR. Two rodent models for depression-like behavior show reduced brain ZDHHC21 expression and attenuated 5-HT1AR palmitoylation. Moreover, selective knock-down of ZDHHC21 in the murine forebrain induced depression-like behavior. We also identified the microRNA miR-30e as a negative regulator of Zdhhc21 expression. Through analysis of the post-mortem brain samples in individuals with MDD that died by suicide we find that miR-30e expression is increased, while ZDHHC21 expression, as well as palmitoylation of 5-HT1AR, are reduced within the prefrontal cortex. Our study suggests that downregulation of 5-HT1AR palmitoylation is a mechanism involved in depression, making the restoration of 5-HT1AR palmitoylation a promising clinical strategy for the treatment of MDD.


Assuntos
Encéfalo/fisiopatologia , Depressão/fisiopatologia , Transtorno Depressivo Maior/fisiopatologia , Receptor 5-HT1A de Serotonina/metabolismo , Aciltransferases/genética , Aciltransferases/metabolismo , Animais , Encéfalo/metabolismo , Linhagem Celular Tumoral , Depressão/genética , Depressão/metabolismo , Transtorno Depressivo Maior/genética , Regulação da Expressão Gênica , Humanos , Lipoilação , Masculino , Camundongos Endogâmicos C57BL , MicroRNAs/genética , Ratos Wistar , Receptor 5-HT1A de Serotonina/genética
7.
J Cell Sci ; 132(16)2019 08 23.
Artigo em Inglês | MEDLINE | ID: mdl-31371490

RESUMO

Protein-protein interaction is often investigated using quantitative molecular microscopy with Förster resonant energy transfer (FRET). Here, we combined 'linear unmixing FRET' (lux-FRET) with the simultaneous application of a FRET-based biosensor for cAMP to investigate the oligomerization between the 5-HT7 receptor (5-HT7R, also known as HTR7) and the 5-HT1A receptor (5-HT1AR, also known as HTR1A) and its importance for cAMP signaling. We found that the 5-HT7R not only stimulates cAMP production, but also forms hetero-oligomers with 5-HT1AR, which blocks the inhibitory effect of the latter. 5-HT7R signaling, however, is not affected by this hetero-oligomerization. By modeling the kinetics of intracellular cAMP level changes in relation to the 5-HT7R:5-HT1AR stoichiometry, we were able to decipher the complex signaling characteristics of endogenous serotonin receptors in cultured hippocampal neurons. Our findings indicate that serotonergic signaling is not only modulated by the concentration of an individual receptor but also by its specific interaction with other receptors in endogenous systems. We conclude that the regulated ratio of serotonin receptors in immature and mature neurons may be critically involved in both the onset and response to treatments of psychiatric diseases, such as anxiety and depression.


Assuntos
AMP Cíclico/metabolismo , Multimerização Proteica , Receptor 5-HT1A de Serotonina/metabolismo , Receptores de Serotonina/metabolismo , Sistemas do Segundo Mensageiro , Animais , Linhagem Celular Tumoral , AMP Cíclico/genética , Camundongos , Receptor 5-HT1A de Serotonina/genética , Receptores de Serotonina/genética
8.
Psychiatr Danub ; 31(2): 256-262, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31291234

RESUMO

BACKGROUND: Post-traumatic stress disorder (PTSD) is a stress related disorder which can occur in an individual after exposure to a traumatic event. It most commonly co-occurs with depression. The two disorders share not only overlapping symptoms, but also genetic diathesis. The aim of this study was to investigate the potential role of single nucleotide polymorphisms (SNPs) of the two serotonergic candidate genes 5-hydroxytryptamine receptor 1A (HTR1A) and tryptophan hydroxylase 2 (TPH2) in the pathogenesis of PTSD and comorbid psychopathology. SUBJECTS AND METHODS: 719 (487 males, 232 females) participants who had experienced war-related trauma between 1991 and 1999 in Bosnia and Herzegovina, Kosovo and Croatia were included in the study. The Sociodemographic questionnaire, Mini International Neuropsychiatric Interview (M.I.N.I.), Clinician Administered PTSD Scale (CAPS) and Brief Symptom Inventory (BSI) were used to collect clinical data. The SNPs rs6295 (HTR1A), rs11178997 and rs1386494 (TPH2) were investigated for their association with PTSD and comorbid psychopathology. RESULTS: A nominal significant association was found between the BSI total score in Lifetime PTSD with the SNP rs6295 of the HTR1A gene. The best result was seen in the dominant model (P=0.018), with the minor allele (C) being the risk allele. Several BSI subscores were also associated with the minor (C) allele in Lifetime PTSD. No association was found for the TPH2 SNPs rs11178997 and rs1386494 in relation to PTSD or comorbid psychopathology. CONCLUSIONS: Our findings suggest that rs6295 in the HTR1A gene may contribute to the psychopathology of PTSD.


Assuntos
Alelos , Receptor 5-HT1A de Serotonina/genética , Transtornos de Estresse Pós-Traumáticos/genética , Triptofano Hidroxilase/genética , Conflitos Armados/psicologia , Bósnia e Herzegóvina , Croácia , Feminino , Humanos , Kosovo , Masculino , Pessoa de Meia-Idade
9.
Behav Brain Res ; 373: 112062, 2019 11 05.
Artigo em Inglês | MEDLINE | ID: mdl-31288061

RESUMO

Anxiety disorders affect nearly twice as many women as men. However, little is known regarding sex-dependent developmental behavioral differences and whether there is an association with later life anxiety disorders. The present study assessed the developmental-behavioral milestones (DBMs) and their relationship with later life anxiety-like behaviors by comparing postnatal ultrasonic vocalizations (USVs) with open field (OF), elevated plus maze (EPM), and light/dark (LD) anxiety test outcomes using the serotonin 1A receptor knockout (KO) mouse model of anxiety. The USVs and DBMs (i.e., grasping, righting, and startle reflexes) were examined on postnatal day 6 (P6), P8, and P10. Adult anxiety-like behaviors were examined on P60 to compare the genotype and sex-dependent differences in anxiety-like behaviors and to correlate them with the USVs. The total number of USVs observed on P8 correlated with later life anxiety-like behaviors in a genotype-, age-, and sex-dependent manner. Interestingly, female KO (KOF) mice exhibited elevated levels of anxiety-like behavior within the OF, EPM, and LD tests. Additionally, an investigation of the USV subtypes, as well as USV sequence structure and repertoire variation, revealed that the KOF mice produced less complex USVs and complex USV-containing sequences on P10. The present study provides an intriguing, predictive "P8/10-USV-to-P60" anxiety-like behavioral model that may prove useful in future characterization, psychopharmacology, and drug rescue studies directed towards sex-specific anxiety treatment.


Assuntos
Ansiedade/fisiopatologia , Receptor 5-HT1A de Serotonina/metabolismo , Vocalização Animal/fisiologia , Fatores Etários , Animais , Ansiedade/genética , Transtornos de Ansiedade/fisiopatologia , Biomarcadores , Feminino , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Fenótipo , Receptor 5-HT1A de Serotonina/genética , Fatores Sexuais , Ondas Ultrassônicas , Ultrassom
10.
Brain Res ; 1719: 243-252, 2019 09 15.
Artigo em Inglês | MEDLINE | ID: mdl-31194947

RESUMO

Serotonin (5-HT) in the central nervous system regulates a variety of biological functions, from the basic homeostatic control to higher brain functions, by acting on fourteen known receptor subtypes. However, it is still usually unclear which receptor subtype is responsible for a specific function due to the lack of highly selective ligands for most of these receptors. Although 5-HT receptor knockout mice are useful, the brain-wide distribution of various receptors makes it difficult to dissect receptor functions in specific and brain regions and cell types. Recent advances in CRISPR/Cas9-mediated in vivo genome editing technology may overcome this problem. In this study, we constructed a viral vector expressing a single guide (sg)RNA targeting Htr1a (sgHtr1a) and Cre recombinase under the control of a neuron-specific promoter. Injection of the viral vector into the dorsal raphe nucleus (DRN) of Cre-dependent Cas9 knock-in mice induced Cre-dependent Cas9 expression mainly in DRN serotonin and GABA neurons. Mismatch cleavage assay and Sanger sequencing showed insertion or deletion formation at the target site. 5-HT1A receptor agonist-induced hypothermia was attenuated and antidepressant effect of a selective serotonin reuptake inhibitor (SSRI) was enhanced by microinjection of the viral vector expressing sgHtr1a into the DRN of Cre-dependent Cas9 knock-in mice. These results suggest that this in vivo CRISPR/Cas9-mediated 5-HT receptor gene knockout strategy provides a reliable and low-cost method for elucidating 5-HT receptor functions in specific cell types and brain regions. Further, we demonstrate that the neuronal 5-HT1A receptor in the DRN regulates body temperature and antidepressant effect of SSRI.


Assuntos
Regulação da Temperatura Corporal/genética , Núcleo Dorsal da Rafe/fisiologia , Receptor 5-HT1A de Serotonina/genética , Animais , Regulação da Temperatura Corporal/fisiologia , Sistemas CRISPR-Cas/genética , Núcleo Dorsal da Rafe/metabolismo , Feminino , Edição de Genes/métodos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Inibição Neural/fisiologia , Receptor 5-HT1A de Serotonina/metabolismo , Receptores de Serotonina/metabolismo , Neurônios Serotoninérgicos/metabolismo , Serotonina/metabolismo , Antagonistas da Serotonina/farmacologia , Agonistas do Receptor de Serotonina/farmacologia , Inibidores de Captação de Serotonina/farmacologia
11.
Mol Cell Endocrinol ; 493: 110472, 2019 08 01.
Artigo em Inglês | MEDLINE | ID: mdl-31167113

RESUMO

Serotonin (5-HT) acts as a morphogen influencing embryonic brain development, and as a neurotransmitter regulating multiple biological functions with lifelong effects on animal physical, physiological and mental health, especially during the rapid growth phase prior to birth when embryos face many challenges to reach structural and functional completion. In this study, the development of the serotoninergic (5-HTergic) system and its modulatory effect on the dopaminergic (DAergic) system and related neural circuits were investigated during the mid-late embryogenesis, embryonic day (E)12-E20, in the chicken's brain. During 5-HTergic neuronal maturation, a growth-related anatomical and functional remodeling was highlighted: the 5-HT neurons continuously grew during E12-E20 except for a remarkable regression during E14-E16. Correspondingly, there was a time-dependent change in the 5-HT synthetic capacity. Specifically, 5-HT concentrations in the raphe nuclei increased from E12 to E14, reaching a first plateau during E14-E16, then continuously increased up to E19, and reaching a second plateau between E19-E20. The second plateau of the 5-HT concentration was in correspondence with the establishment of the 5-HTergic autoregulatory loop during E19-E20 and the development of the DAergic system. The DA concentrations remained unchanged from E12 to E16, then started to increase at E16, reaching a maximum at E19, and diminished before hatching. The unique developing time sequence between the 5-HTergic and DAergic systems suggests that the 5-HTergic system may play a critical role in forming the 5-HT - DA neural circuit during chicken embryogenesis. These results provide new insights for understanding the functional organization of the 5-HTergic system during embryonic development and raise the possibility that prenatally modulating the 5-HTergic system may lead to long-lasting brain structural and functional alterations.


Assuntos
Encéfalo/embriologia , Dopamina/metabolismo , Serotonina/metabolismo , Animais , Encéfalo/metabolismo , Galinhas , Cromatografia Líquida de Alta Pressão , Desenvolvimento Embrionário , Regulação da Expressão Gênica no Desenvolvimento , Receptor 5-HT1A de Serotonina/genética , Receptor 5-HT1A de Serotonina/metabolismo , Proteínas da Membrana Plasmática de Transporte de Serotonina/genética , Proteínas da Membrana Plasmática de Transporte de Serotonina/metabolismo , Triptofano Hidroxilase/genética , Triptofano Hidroxilase/metabolismo
12.
Asian J Psychiatr ; 43: 184-188, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31228794

RESUMO

INTRODUCTION: Major Depressive Disorder (MDD) is a broad heterogeneous diagnostic construct. Previous studies have shown that it can be resolved into several symptom-clusters which are proposed to be associated with single nucleotide polymorphisms (SNPs) of the serotonergic pathway (5-HTTLPR, 5HTR1A, 5-HTR2A). METHODS AND MATERIAL: In a cross-sectional study conducted at a tertiary level mental health care set-up in north India, 80 out-patients with MDD were evaluated with Montgomery Asberg Depression Rating Scale (MADRS) and then genotyping was done. The different clinical and genetic variables were compared across the factor structures of MADRS. Also, the comparison of the genetic data of cases was done with the pre-existing database of the non-blood related healthy ethnically-matched controls. RESULTS: There was no significant association between age, gender, other clinical variables, SNPs like 5-HTTLPR SS/SL, rs6295 CC/CG/GG, rs6311GG/GA/AA, rs6313 CC/CT/TT and different factor-structures like 'detachment' consisting of items like concentration difficulty, lassitude, inability to feel; 'psychic anxiety' consisting of suicidal thoughts and inner tension; 'mood-pessimism' consisting of symptoms like apparent sadness, reported sadness, pessimistic thoughts and 'vegetative symptoms' like decreased sleep, poor appetite. Neither there was any association between genotype of the cases compared with the controls. CONCLUSIONS: No significant association was obtained between the four-factor structures of depression in MADRS and serotonin transporter and receptor SNPs in a study with a small sample size. This study evaluates whether depression symptom-clusters have distinct genotypic determinants and necessitates more comprehensive studies for unravelling the genetic determinants of depression.


Assuntos
Transtorno Depressivo Maior/genética , Transtorno Depressivo Maior/fisiopatologia , Receptor 5-HT1A de Serotonina/genética , Receptor 5-HT2A de Serotonina/genética , Proteínas da Membrana Plasmática de Transporte de Serotonina/genética , Adulto , Estudos Transversais , Transtorno Depressivo Maior/classificação , Feminino , Humanos , Índia , Masculino , Pessoa de Meia-Idade , Polimorfismo de Nucleotídeo Único
13.
Mol Biol Evol ; 36(7): 1418-1429, 2019 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-31045220

RESUMO

Serotonin is a neurotransmitter that plays an important role in regulating behavior and personality in humans and other mammals. Polymorphisms in genes coding for the serotonin receptor subtype 1A (HTR1A), the serotonin transporter (SLC6A4), and the serotonin degrading enzyme monoamine oxidase A (MAOA) are associated with anxiety, impulsivity, and neurotic personality in humans. In primates, previous research has largely focused on SLC6A4 and MAOA, with few studies investigating the role of HTR1A polymorphic variation on behavior. Here, we examined variation in the coding region of HTR1A across apes, and genotyped polymorphic coding variation in a sample of 214 chimpanzees with matched measures of personality and behavior. We found evidence for positive selection at three amino acid substitution sites, one in chimpanzees-bonobos (Thr26Ser), one in humans (Phe33Val), and one in orangutans (Ala274Gly). Investigation of the HTR1A coding region in chimpanzees revealed a polymorphic site, where a C/A single nucleotide polymorphism changes a proline to a glutamine in the amino acid sequence (Pro248Gln). The substitution is located in the third intracellular loop of the receptor, a region important for serotonin signal transduction. The derived variant is the major allele in this population (frequency 0.67), and is associated with a reduction in anxiety, decreased rates of male agonistic behavior, and an increase in socio-positive behavior. These results are the first evidence that the HTR1A gene may be involved in regulating social behavior in chimpanzees and encourage further systematic investigation of polymorphic variation in other primate populations with corresponding data on behavior.


Assuntos
Comportamento Agonístico , Ansiedade/genética , Pan troglodytes/genética , Receptor 5-HT1A de Serotonina/genética , Sequência de Aminoácidos , Animais , Feminino , Variação Genética , Masculino , Pan troglodytes/psicologia , Personalidade/genética
14.
ACS Chem Neurosci ; 10(7): 3154-3166, 2019 07 17.
Artigo em Inglês | MEDLINE | ID: mdl-31140276

RESUMO

Serotonin (5-hydroxytryptamine; 5-HT) coordinates behavioral responses to stress through a variety of presynaptic and postsynaptic receptors distributed across functionally diverse neuronal networks in the central nervous system. Efferent 5-HT projections from the dorsal raphe nucleus (DRN) to the bed nucleus of the stria terminalis (BNST) are generally thought to enhance anxiety and aversive learning by activating 5-HT2C receptor (5-HT2CR) signaling in the BNST, although an opposing role for postsynaptic 5-HT1A receptors has recently been suggested. In the present study, we sought to delineate a role for postsynaptic 5-HT1A receptors in the BNST in aversive behaviors using a conditional knockdown of the 5-HT1A receptor. Both males and females were tested to dissect out sex-specific effects. We found that male mice have significantly reduced fear memory recall relative to female mice and inactivation of 5-HT1A receptor in the BNST increases contextual fear conditioning in male mice so that they resemble the females. This coincided with an increase in neuronal excitability in males, suggesting that 5-HT1A receptor deletion may enhance contextual fear recall by disinhibiting fear memory circuits in the BNST. Interestingly, 5-HT1A receptor knockdown did not significantly alter anxiety-like behavior in male or female mice, which is in agreement with previous findings that anxiety and fear are modulated by dissociable circuits in the BNST. Overall, these results suggest that BNST 5-HT1A receptors do not significantly alter behavior under basal conditions, but can act as a molecular brake that buffer against excessive activation of aversive circuits in more threatening contexts.


Assuntos
Ansiedade/metabolismo , Medo/fisiologia , Neurônios/metabolismo , Receptor 5-HT1A de Serotonina/metabolismo , Núcleos Septais/metabolismo , Animais , Comportamento Animal/fisiologia , Comportamento Alimentar/fisiologia , Feminino , Técnicas de Silenciamento de Genes , Masculino , Camundongos , Camundongos Transgênicos , Atividade Motora/fisiologia , Receptor 5-HT1A de Serotonina/genética , Fatores Sexuais
15.
Mol Neurobiol ; 56(11): 7680-7693, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31098953

RESUMO

Disturbed serotonergic signaling in the hippocampus observed in many individuals vulnerable to stress has been suggested as one of the primary factors contributing to the development of depression. However, little is known about the physiology of the brain in the resilient phenotype. Resilient subjects maintain a positive mood and psychological balance despite being under the stress influence. In our study, we generated stress-vulnerable and resilient rats by using a chronic mild stress (CMS) paradigm. Using different molecular approaches, we revealed that resilient animals exhibited a significantly decreased expression level of miR-18a-5p and, in the same time, an elevated level of 5-HT1AR in dorsal, but not ventral, part of the hippocampus. Described biochemical changes were not observed in animals behaviorally vulnerable to stress. Further, in vitro analysis showed that miR-18a-5p may be a negative epigenetic regulator of 5-HT1AR since the treatment of adult hippocampal neurons with miR-18a-5p mimic significantly lowered the expression level of mRNA encoding 5-HT1AR. Moreover, bioinformatic analysis of potential target genes expressed in the hippocampus and being regulated by miR-18a-5p showed that this microRNA may regulate biological processes, such as axonogenesis, which are important in the functioning of the hippocampus in both rats and humans. All these molecular features may contribute to serotonergic homeostatic balance at the level of serotonin turnover observed in hippocampi of resilient but not stress-vulnerable rats. Delineation of further molecular and biochemical markers underlying resilience to stress may contribute to the development of new antidepressant strategies which will restore resilient phenotype in depressed patients.


Assuntos
Hipocampo/metabolismo , MicroRNAs/genética , Receptor 5-HT1A de Serotonina/genética , Resiliência Psicológica , Estresse Psicológico/genética , Animais , Encéfalo/metabolismo , Corticosterona/sangue , Regulação da Expressão Gênica , Ácido Hidroxi-Indolacético/metabolismo , Masculino , MicroRNAs/metabolismo , Fenótipo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos , Receptor 5-HT1A de Serotonina/metabolismo , Receptores de Esteroides/sangue , Sacarose
16.
Neuropharmacology ; 153: 134-141, 2019 07 15.
Artigo em Inglês | MEDLINE | ID: mdl-31078489

RESUMO

Brain serotonin (5-HT) plays a key role in aggressive behaviours and related psychopathologies, but its precise mechanism of action remains elusive. Genetic animal models may provide a tool to elucidate the relationship between aggression and serotonin. The present study showed that tryptophan hydroxylase 2 (Tph2) knockout (KO) rats, which exhibit profoundly diminished extracellular serotonin levels, display increased aggressiveness compared to their Tph2 wildtype (WT) counterparts. However, the level of aggression in Tph2 KO rats did not equal that of feral wild type Groningen (WTG) rats. To investigate whether enhanced 5-HT1A receptor functionality may be present in Tph2 KO rats, we tested the acute anti-aggressive potency of the highly selective 5-HT1A receptor full agonist NLX-112 (a.k.a. befiradol or F13640). Data show that compared to Tph2 WT and WTG rats, the NLX-112 dose-effect curve was shifted to the right in Tph2 KO animals. These results suggest that, unlike previous reports in Tph2 KO mice, Tph2 KO rats have a decreased 5-HT1A receptor sensitivity compared to both Tph2 WT and WTG animals.


Assuntos
Agressão/fisiologia , Fenótipo , Receptor 5-HT1A de Serotonina/metabolismo , Agonistas do Receptor 5-HT1 de Serotonina/farmacologia , Triptofano Hidroxilase/deficiência , Agressão/efeitos dos fármacos , Agressão/psicologia , Animais , Relação Dose-Resposta a Droga , Feminino , Masculino , Distribuição Aleatória , Ratos , Ratos Transgênicos , Receptor 5-HT1A de Serotonina/genética , Triptofano Hidroxilase/genética
17.
Food Funct ; 10(5): 2720-2728, 2019 May 22.
Artigo em Inglês | MEDLINE | ID: mdl-31033966

RESUMO

Piperine is the crucial alkaloid component of black pepper (Piper nigrum Linn.) and has neuroprotective effects. Because inhibition of glutamatergic excitatory neurotransmission is a possible mechanism involved in neuroprotection, we investigated the effect of piperine on the 4-aminopyridine (4-AP)-evoked release of glutamate from rat hippocampal synaptosomes. Piperine inhibited 4-AP-evoked glutamate release, and the inhibition was prevented by the chelation of extracellular Ca2+ ions and a vesicular transporter inhibitor. Piperine reduced the 4-AP-evoked elevation of intrasynaptosomal Ca2+ levels but did not affect the synaptosomal membrane potential. In the presence of ω-conotoxin MVIIC, an N- and P/Q-type channel blocker, the piperine-mediated inhibition of 4-AP-evoked glutamate release was markedly reduced; however, dantrolene and CGP37157, which are intracellular Ca2+-release inhibitors, did not alter the piperine effect. In addition, immunocytochemical analysis confirmed the presence of presynaptic 5-hydroxytryptamine 1A (5-HT1A) receptor proteins. The glutamate release-inhibiting effect of piperine was discovered to be prevented by the 5-HT1A receptor antagonist WAY100635 and the G protein ßγ subunit inhibitor gallein; however, it was unaffected by the adenylate cyclase inhibitor SQ22536 or the protein kinase A inhibitor PKI622. These results suggest that piperine inhibits glutamate release from rat hippocampal nerve terminals by reducing Ca2+ influx through N- and P/Q-type Ca2+ channels and that the activation of presynaptic 5-HT1A receptors and the G protein ßγ subunit is involved in this effect.


Assuntos
Alcaloides/farmacologia , Benzodioxóis/farmacologia , Cálcio/metabolismo , Subunidades beta da Proteína de Ligação ao GTP/metabolismo , Subunidades gama da Proteína de Ligação ao GTP/metabolismo , Ácido Glutâmico/metabolismo , Hipocampo/metabolismo , Piperidinas/farmacologia , Alcamidas Poli-Insaturadas/farmacologia , Receptor 5-HT1A de Serotonina/metabolismo , 4-Aminopiridina/metabolismo , Animais , Canais de Cálcio/genética , Canais de Cálcio/metabolismo , Subunidades beta da Proteína de Ligação ao GTP/genética , Subunidades gama da Proteína de Ligação ao GTP/genética , Hipocampo/efeitos dos fármacos , Masculino , Piper nigrum/química , Terminações Pré-Sinápticas/efeitos dos fármacos , Terminações Pré-Sinápticas/metabolismo , Ratos , Ratos Sprague-Dawley , Receptor 5-HT1A de Serotonina/genética , Sinaptossomos/efeitos dos fármacos , Sinaptossomos/metabolismo
18.
Biochem Biophys Res Commun ; 511(2): 440-446, 2019 04 02.
Artigo em Inglês | MEDLINE | ID: mdl-30808545

RESUMO

The association between dysregulated serotonergic activity and major depressive disorder (MDD) is well known. However, the various mechanisms underlying serotonergic dysregulation in MDD remain unclear. Previous research on serotonergic (5-HT) neurons identified microRNA-26a (miR-26a) targeting of the serotonin autoreceptor, 5-HT receptor 1A (HTR1A). Reporter assays with the Htr1a 5'UTR sequence were performed in vitro. Adult transgenic mouse models altering miR-26a-2 and Htr1a expression were used for chronic social defeat, antidepressant treatment, and in vivo lentiviral experiments. Mice were tested for anxiety-like behavior using the elevated plus-maze, dark-light transfer, and open-field tests, and for depression-like behavior using the forced-swim test. We confirmed that miR-26a-2 downregulates Htr1a expression in 5-HT neurons in vitro. miR-26a-2 levels were significantly upregulated in the mouse dorsal raphe nucleus (DRN) following antidepressant therapy. The transgenic murine model overexpressing miR-26a-2 in serotonergic neurons displayed improved behavioral resiliency to social defeat. These effects were abrogated by the addition of Htr1a overexpression. In contrast, the transgenic murine model with miR-26a-2 knockdown in serotonergic neurons displayed increased anxious behavior and weakened antidepressant response. These effects were rescued by silencing Htr1a expression. Our findings suggest that miR-26a-2 functions as an endogenous antidepressant by targeting HTR1A in serotonergic neurons.


Assuntos
Antidepressivos/uso terapêutico , Transtorno Depressivo Maior/tratamento farmacológico , Transtorno Depressivo Maior/genética , MicroRNAs/genética , Receptor 5-HT1A de Serotonina/genética , Animais , Ansiedade/complicações , Ansiedade/genética , Ansiedade/patologia , Transtorno Depressivo Maior/complicações , Transtorno Depressivo Maior/patologia , Modelos Animais de Doenças , Regulação para Baixo/efeitos dos fármacos , Camundongos , Camundongos Transgênicos , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Neurônios/patologia , Regulação para Cima/efeitos dos fármacos
19.
J Psychiatry Neurosci ; 44(3): 164-176, 2019 05 01.
Artigo em Inglês | MEDLINE | ID: mdl-30807072

RESUMO

Major depression and anxiety are highly prevalent and involve chronic dysregulation of serotonin, but they remain poorly understood. Here, we review novel transcriptional (genetic, epigenetic) and posttranscriptional (microRNA, alternative splicing) mechanisms implicated in mental illness, focusing on a key serotonin-related regulator, the serotonin 1A (5-HT1A) receptor. Functional single-nucleotide polymorphisms and stress-induced DNA methylation of the 5-HT1A promoter converge to differentially alter pre- and postsynaptic 5-HT1A receptor expression associated with major depression and reduced therapeutic response to serotonergic antidepressants. Major depression is also associated with altered levels of splice factors and microRNA, posttranscriptional mechanisms that regulate RNA stability. The human 5-HT1A 3'-untranslated region is alternatively spliced, removing microRNA sites and increasing 5-HT1A expression, which is reduced in major depression and may be genotype-dependent. Thus, the 5-HT1A receptor gene illustrates the convergence of genetic, epigenetic and posttranscriptional mechanisms in gene expression, neurodevelopment and neuroplasticity, and major depression. Understanding gene regulatory mechanisms could enhance the detection, categorization and personalized treatment of major depression.


Assuntos
Processamento Alternativo/genética , Metilação de DNA/genética , Transtorno Depressivo Maior/genética , Epigênese Genética/genética , MicroRNAs/genética , Receptor 5-HT1A de Serotonina/genética , Transcrição Genética/genética , Animais , Transtorno Depressivo Maior/tratamento farmacológico , Humanos
20.
Artigo em Inglês | MEDLINE | ID: mdl-30690153

RESUMO

Serotonin is involved in the regulation of many physiological and behavioral processes in vertebrates and invertebrates. The effects of serotonin are mediated through interactions of several 5-HT receptor types. The expression and pharmacological properties of 5-HT1 have received more attention than other serotonin receptors, but its functions at the individual level are little studied in arthropods. Silkworm, a Lepidoptera model, almost has no reports about serotonin receptors. To analyze the function of Bm5-HT1A receptor in vitro, the ORF of Bm5-HT1A was cloned into the pcDNA3.1 vector and expressed in HEK 293 cells. Serotonin activation of Bm5-HT1A-expressing cells decreased forskolin-stimulated cAMP synthesis and had the most potent effect compared to other biogenic amines. Serotonin reduced cAMP synthesis in a dose-dependent manner, and half-maximal activation (EC50) occurred at a concentration of 1.17 × 10-7 M (117 nM). The pharmacological analysis demonstrated that the rank potency of agonists was pimozide >8-OH-DPAT >5-MeOT ~ αm-5-HT, and antagonists was WAY-100635 > prazosin > SB-269970 > methiothepin at the Bm5-HT1A receptor. Injecting the antagonist of Bm5-HT1A receptor into larvae caused slow or weak motility, and adults lowered courtship vitality or moving speed. Injecting dsRNA of Bm5-HT1A into adults also dropped locomotivity in courtship. These results show that the Bm5-HT1A receptor is related to locomotor activity. This study provides the first information of serotonin receptor on pharmacological in silkworm and on individual functions in arthropods.


Assuntos
Bombyx/fisiologia , Locomoção/fisiologia , Receptor 5-HT1A de Serotonina/fisiologia , Animais , Relação Dose-Resposta a Droga , Feminino , Células HEK293 , Humanos , Masculino , Receptor 5-HT1A de Serotonina/efeitos dos fármacos , Receptor 5-HT1A de Serotonina/genética , Serotonina/farmacologia
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