RESUMO
Gardenia jasminoides Ellis is a famous fragrant flower in China. Previous pharmacological research mainly focuses on its fruit. In this study, the essential oil of the flower of 'Shanzhizi', which was a major variety for traditional Chinese medicine use, was extracted by hydro distillation and analyzed by GC-MS. Mouse anxiety models included open field, elevated plus maze (EPM), and light and dark box (LDB), which were used to evaluate its anxiolytic effect via inhalation. The involvement of monoamine system was studied by pretreatment with neurotransmitter receptor antagonists WAY100635, flumazenil and sulpiride. The monoamine neurotransmitters contents in the prefrontal cortex (PFC) and hippocampus after aroma inhalation were also analyzed. The results showed that inhalation of G. jasminoides essential oil could significantly elevated the time and entries into open arms in EPM tests and the time explored in the light chamber in LDB tests with no sedative effect. WAY100635 and sulpiride, but not flumazenil, blocked its anxiolytic effect. Inhalation of G. jasminoides essential oil significantly down-regulated the 5-HIAA/5-HT in the PFC and reduced the 5-HIAA content in hippocampus compared to the control treatment. In conclusion, inhalation of gardenia essential oil showed an anxiolytic effect in mice. Monoamine, especially the serotonergic system, was involved in its anxiolytic effect.
Assuntos
Ansiolíticos/farmacologia , Gardenia/química , Óleos Voláteis/química , Óleos Voláteis/farmacologia , Administração por Inalação , Animais , Ansiolíticos/administração & dosagem , Ansiolíticos/química , Monoaminas Biogênicas/análise , Cicloexanos/farmacologia , Modelos Animais de Doenças , Medicamentos de Ervas Chinesas/química , Medicamentos de Ervas Chinesas/farmacologia , Teste de Labirinto em Cruz Elevado , Flumazenil/farmacologia , Cromatografia Gasosa-Espectrometria de Massas , Hipnóticos e Sedativos/administração & dosagem , Hipnóticos e Sedativos/química , Hipnóticos e Sedativos/farmacologia , Masculino , Camundongos Endogâmicos ICR , Óleos Voláteis/administração & dosagem , Pentobarbital/farmacologia , Piperazinas/farmacologia , Córtex Pré-Frontal/efeitos dos fármacos , Córtex Pré-Frontal/metabolismo , Receptores de Neurotransmissores/antagonistas & inibidores , Sono/efeitos dos fármacos , Sulpirida/farmacologia , Transmissão Sináptica/efeitos dos fármacosRESUMO
Neurotransmitters are conventionally viewed as nerve-secreted substances that mediate the stimulatory or inhibitory neuronal functions through binding to their respective receptors. In the past decades, many novel discoveries come to light elucidating the regulatory roles of neurotransmitters in the physiological and pathological functions of tissues and organs. Notably, emerging data suggest that cancer cells take advantage of the neurotransmitters-initiated signaling pathway to activate uncontrolled proliferation and dissemination. In addition, neurotransmitters can affect immune cells and endothelial cells in the tumor microenvironment to promote tumor progression. Therefore, a better understanding of the mechanisms underlying neurotransmitter function in tumorigenesis, angiogenesis, and inflammation is expected to enable the development of the next generation of antitumor therapies. Here, we summarize the recent important studies on the different neurotransmitters, their respective receptors, target cells, as well as pro/antitumor activity of specific neurotransmitter/receptor axis in cancers and provide perspectives and insights regarding the rationales and strategies of targeting neurotransmitter system to cancer treatment.
Assuntos
Inflamação/patologia , Neoplasias/patologia , Neovascularização Patológica/patologia , Neurotransmissores/metabolismo , Receptores de Neurotransmissores/metabolismo , Animais , Antineoplásicos/uso terapêutico , Carcinogênese/efeitos dos fármacos , Carcinogênese/patologia , Modelos Animais de Doenças , Humanos , Inflamação/tratamento farmacológico , Inflamação/imunologia , Camundongos , Neoplasias/irrigação sanguínea , Neoplasias/tratamento farmacológico , Neoplasias/imunologia , Neovascularização Patológica/tratamento farmacológico , Neovascularização Patológica/imunologia , Neurotransmissores/antagonistas & inibidores , Receptores de Neurotransmissores/antagonistas & inibidores , Transdução de Sinais/efeitos dos fármacos , Microambiente Tumoral/efeitos dos fármacos , Microambiente Tumoral/imunologiaRESUMO
We investigated age-related changes in in vivo and in vitro functions and gene expression of the bladder of male and female mice. Mature and aged (12 and 27-30 month old) C57BL/6 mice of both sexes were used. Frequency volume, conscious free-moving cystometry and detrusor contractile and relaxant properties in in vitro organ bath were evaluated. mRNA expression level of muscarinic, purinergic, and ß-adrenergic receptors and gene expression changes by cDNA microarray analysis of the bladder were determined. Cystometry demonstrated storage and voiding dysfunctions with ageing in both sexes. Detrusor strips from aged mice showed weaker contractile responses particularly in the cholinergic component and weaker relaxant responses to isoproterenol. These age-related impairments were generally severer in males. mRNA expression of bladder tissue was decreased for M3 muscarinic receptors in aged males and ß2-adrenoceptors in aged females. cDNA microarray analysis results, albeit substantial sex difference, indicated "cell-to-cell signaling and interaction" as the most common feature of age-related gene expression. In summary, aged mice demonstrated voiding and storage dysfunctions resembling to detrusor hyperactivity with impaired contractility (DHIC), which were more pronounced in males. Genomic changes associated with aging may contribute to the age-related bladder functional deterioration in mice.
Assuntos
Envelhecimento/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Contração Muscular , Bexiga Urinária/metabolismo , Envelhecimento/fisiologia , Animais , Feminino , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Receptores de Neurotransmissores/antagonistas & inibidores , Receptores de Neurotransmissores/genética , Receptores de Neurotransmissores/metabolismo , Bexiga Urinária/crescimento & desenvolvimento , Bexiga Urinária/fisiologiaRESUMO
Rabies virus induces drastic behaviour modifications in infected hosts. The mechanisms used to achieve these changes in the host are not known. The main finding of this study is that a region in the rabies virus glycoprotein, with homologies to snake toxins, has the ability to alter behaviour in animals through inhibition of nicotinic acetylcholine receptors present in the central nervous system. This finding provides a novel aspect to virus receptor interaction and host manipulation by pathogens in general. The neurotoxin-like region of the rabies virus glycoprotein inhibited acetylcholine responses of α4ß2 nicotinic receptors in vitro, as did full length ectodomain of the rabies virus glycoprotein. The same peptides significantly altered a nicotinic receptor induced behaviour in C. elegans and increased locomotor activity levels when injected into the central nervous system of mice. These results provide a mechanistic explanation for the behavioural changes in hosts infected by rabies virus.
Assuntos
Sistema Nervoso Central/metabolismo , Sistema Nervoso Central/virologia , Glicoproteínas/química , Interações Hospedeiro-Patógeno , Vírus da Raiva/fisiologia , Receptores de Neurotransmissores/antagonistas & inibidores , Venenos de Serpentes/química , Sequência de Aminoácidos , Animais , Comportamento Animal , Caenorhabditis elegans/virologia , Sequência Conservada , Humanos , Camundongos , Neurotoxinas/química , Neurotoxinas/metabolismo , Peptídeos/química , Peptídeos/metabolismo , Ligação Proteica , Domínios Proteicos , Receptores de Neurotransmissores/metabolismo , Receptores Nicotínicos/química , Receptores Nicotínicos/metabolismo , Homologia de Sequência de Aminoácidos , XenopusRESUMO
Neuromedin U (NMU) mediates various physiological functions via NMUR1 and NMUR2 receptors. NMUR2 has been considered a promising treatment option for diabetes and obesity. Although NMU-8, a shorter peptide, has potent agonist activity for both receptors, it is metabolically unstable. Therefore, NMU-8 analogs modified with long-chain alkyl moieties via a linker were synthesized. An octadecanoyl analog (17) with amino acid substitutions [αMePhe19, Nle21, and Arg(Me)24] and a linker [Tra-γGlu-PEG(2)] dramatically increased NMUR2 selectivity, with retention of high agonist activity. Subcutaneous administration of 17 induced anorectic activity in C57BL/6J mice. Owing to its high metabolic stability, 17 would be useful in clarifying the physiological role and therapeutic application of NMU.
Assuntos
Depressores do Apetite/metabolismo , Peptídeos/metabolismo , Receptores de Neurotransmissores/metabolismo , Alquilação , Sequência de Aminoácidos , Animais , Depressores do Apetite/química , Depressores do Apetite/farmacologia , Ingestão de Alimentos/efeitos dos fármacos , Humanos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Peptídeos/agonistas , Receptores de Neurotransmissores/antagonistas & inibidores , Relação Estrutura-AtividadeRESUMO
During acute neuroinflammation, increased levels of cytokines within the brain may contribute to synaptic reorganization that results in long-term changes in network hyperexcitability. Indeed, inflammatory cytokines are implicated in synaptic dysfunction in epilepsy and in an array of degenerative and autoimmune diseases of the central nervous system. Current tools for studying the impact of inflammatory factors on neural networks are either insufficiently fast and sensitive or require complicated and costly experimental rigs. Calcium imaging offers a reasonable surrogate for direct measurement of neuronal network activity, but traditional imaging paradigms are confounded by cellular heterogeneity and cannot readily distinguish between glial and neuronal calcium transients. While the establishment of pure neuron cultures is possible, the removal of glial cells ignores physiologically relevant cell-cell interactions that may be critical for circuit level disruptions induced by inflammatory factors. To overcome these issues, we provide techniques and algorithms for image processing and waveform feature extraction using automated analysis of spontaneous and evoked calcium transients in primary murine cortical neuron cultures transduced with an adeno-associated viral vector driving the GCaMP6f reporter behind a synapsin promoter. Using this system, we provide evidence of network perturbations induced by the inflammatory cytokines TNFα, IL1ß, and IFNγ.
Assuntos
Cálcio/metabolismo , Córtex Cerebral/citologia , Córtex Cerebral/patologia , Citocinas/metabolismo , Imagem Molecular , Vias Neurais , Neurônios/fisiologia , Animais , Biomarcadores , Sinalização do Cálcio , Células Cultivadas , Imunofluorescência , Expressão Gênica , Mediadores da Inflamação/metabolismo , Camundongos , Rede Nervosa , Neurônios/efeitos dos fármacos , Receptores de Neurotransmissores/agonistas , Receptores de Neurotransmissores/antagonistas & inibidoresRESUMO
Combinations of Ca2+ channel inhibitors have been proposed as an effective means to prevent excess Ca2+ flux and death of neurons and glia following neurotrauma in vivo. However, it is not yet known if beneficial outcomes such as improved viability have been due to direct effects on intracellular Ca2+ concentrations. Here, the effects of combinations of Lomerizine (Lom), 2,3-dioxo-7-(1H-imidazol-1-yl)6-nitro-1,2,3,4-tetrahydro-1-quinoxalinyl]acetic acid monohydrate (YM872), 3,5-dimethyl-1-adamantanamine (memantine (Mem)) and/or adenosine 5'-triphosphate periodate oxidized sodium salt (oxATP) to block voltage-gated Ca2+ channels, Ca2+ permeable α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors, NMDA receptors and purinergic P2X7 receptors (P2X7R) respectively, on Ca2+ concentration and viability of rat primary mixed cortical (MC) cultures exposed to hydrogen peroxide (H2O2) insult, were assessed. The contribution of ryanodine-sensitive intracellular stores to intracellular Ca2+ concentration was also assessed. Live cell calcium imaging revealed that a 30min H2O2 insult induced a slow increase in intracellular Ca2+, in part from intracellular sources, associated with loss of cell viability by 6h. Most combinations of inhibitors that included oxATP significantly decreased Ca2+ influx and increased cell viability when administered simultaneously with H2O2. However, reductions in intracellular Ca2+ concentration were not always linked to improved cell viability. Examination of the density of specific cell subpopulations demonstrated that most combinations of inhibitors that included oxATP preserved NG2+ non-oligodendroglial cells, but preservation of astrocytes and neurons required additional inhibitors. Olig2+ oligodendroglia and ED-1+ activated microglia/macrophages were not preserved by any of the inhibitor combinations. These data indicate that following H2O2 insult, limiting intracellular Ca2+ entry via P2X7R is generally associated with increased cell viability. Protection of NG2+ non-oligodendroglial cells by Ca2+ channel inhibitor combinations may contribute to observed beneficial outcomes in vivo.
Assuntos
Cálcio/metabolismo , Neuroglia/efeitos dos fármacos , Neurônios/efeitos dos fármacos , Fármacos Neuroprotetores/farmacologia , Neurotransmissores/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Animais , Cátions/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/fisiologia , Córtex Cerebral/efeitos dos fármacos , Córtex Cerebral/metabolismo , Técnicas de Cocultura , Peróxido de Hidrogênio/toxicidade , Imidazóis/farmacologia , Memantina/farmacologia , Neuroglia/metabolismo , Neurônios/metabolismo , Estresse Oxidativo/fisiologia , Piperazinas/farmacologia , Quinoxalinas/farmacologia , Ratos , Receptores de Neurotransmissores/antagonistas & inibidores , Receptores de Neurotransmissores/metabolismoRESUMO
Synthetic derivatives of phenothiazine have been used for over a century as well-tolerated drugs against a variety of human ailments from psychosis to cancer. This implies a considerable diversity in the mechanisms of action produced by structural changes to the phenothiazine scaffold. For example, chlorpromazine treatment of psychosis is related to its interaction with dopaminergic receptors. On the other hand, antagonistic action of such drugs on cholinergic receptor systems would be counter-productive for treatment of Alzheimer's disease. In a search for phenothiazines that are inhibitors of cholinesterases, especially butyrylcholinesterase, with potential to treat Alzheimer's disease, we wished to ascertain that such molecules could be devoid of neurotransmitter receptor interactions. To that end, a number of our synthetic N-10-carbonyl phenothiazine derivatives, with cholinesterase inhibitory activity, were tested for interaction with a variety of neurotransmitter receptor systems. We demonstrate that phenothiazines can be prepared without significant neurotransmitter receptor interactions while retaining high potency as cholinesterase ligands for treatment of Alzheimer's disease.
Assuntos
Butirilcolinesterase/metabolismo , Inibidores da Colinesterase/farmacologia , Fenotiazinas/farmacologia , Receptores de Neurotransmissores/antagonistas & inibidores , Inibidores da Colinesterase/síntese química , Inibidores da Colinesterase/química , Relação Dose-Resposta a Droga , Humanos , Estrutura Molecular , Fenotiazinas/síntese química , Fenotiazinas/química , Receptores de Neurotransmissores/metabolismo , Relação Estrutura-AtividadeRESUMO
Past studies have suggested that progesterone-derived ovarian hormones contribute to the discriminative stimulus effects of ethanol, particularly via progesterone metabolites that act at γ-aminobutyric acid type A (GABA(A)) receptors. It is unknown whether loss of ovarian hormones in women, for example, after menopause, may be associated with altered receptor mediation of the effects of ethanol. The current study measured the substitution of allopregnanolone, pregnanolone, pentobarbital, midazolam, dizocilpine, TFMPP, and RU 24969 in female sham and ovariectomized rats trained to discriminate 1.0 g/kg ethanol from water. The groups did not differ in the substitution of GABA(A)-positive modulators (barbiturates, benzodiazepines, neuroactive steroids) or the N-methyl-D-aspartate receptor antagonist dizocilpine. Similarly, blood-ethanol concentration did not differ between the groups, and plasma adrenocorticotropic hormone, progesterone, pregnenolone, and deoxycorticosterone were unchanged 30 min after administration of 1.0 g/kg ethanol or water. However, substitution of neuroactive steroids and RU 24969, a 5-hydroxytryptamine (5-HT)(1A/1B) receptor agonist, was lower than observed in previous studies of male rats, and TFMPP substitution was decreased in ovariectomized rats. Ovarian hormones appear to contribute to 5-HT receptor mediation of the discriminative stimulus effects of ethanol in rats.
Assuntos
Consumo de Bebidas Alcoólicas/efeitos adversos , Aprendizagem por Discriminação/efeitos dos fármacos , Ciclo Estral/metabolismo , Ovário/fisiologia , Pós-Menopausa/metabolismo , Progesterona/análogos & derivados , Receptores de Progesterona/metabolismo , Dissuasores de Álcool/uso terapêutico , Consumo de Bebidas Alcoólicas/sangue , Consumo de Bebidas Alcoólicas/metabolismo , Consumo de Bebidas Alcoólicas/prevenção & controle , Animais , Comportamento Animal/efeitos dos fármacos , Resistência a Medicamentos , Ciclo Estral/sangue , Etanol/sangue , Feminino , Terapia de Reposição Hormonal , Neurotransmissores/sangue , Neurotransmissores/farmacocinética , Neurotransmissores/uso terapêutico , Ovariectomia/efeitos adversos , Ovário/efeitos dos fármacos , Ovário/metabolismo , Pós-Menopausa/sangue , Progesterona/sangue , Progesterona/farmacocinética , Progesterona/uso terapêutico , Ratos , Ratos Long-Evans , Receptores de Neurotransmissores/agonistas , Receptores de Neurotransmissores/antagonistas & inibidores , Receptores de Neurotransmissores/metabolismo , Receptores de Progesterona/agonistas , Receptores de Progesterona/antagonistas & inibidores , Agonistas do Receptor de Serotonina/sangue , Agonistas do Receptor de Serotonina/farmacocinética , Agonistas do Receptor de Serotonina/uso terapêuticoRESUMO
Acetylcholine, and to a lesser extent ATP, mediates neurogenic contractions of bladder smooth muscle. Recently, the urothelium and lamina propria have also been shown to have contractile properties, but the neurotransmitters involved in mediating responses to nerve stimulation have not been investigated. Isolated strips of porcine urothelium with lamina propria were electrically field stimulated and contractions recorded. Drugs interfering with neurotransmission were then employed to identify which neurotransmitters mediated responses. Strips of urothelium/lamina propria developed spontaneous contractions with a frequency of 3.5±0.1 cycles min⻹ and amplitude of 0.84±0.06 g. Electrical field stimulation at 5, 10, and 20 Hz resulted in frequency-related contractions (1.13±0.36 g, 1.59±0.46 g and 2.20±0.53 g, respectively, n=13), and these were reduced in the presence of tetrodotoxin (1 µm) by 77±20% at 5 Hz, 79±7% at 10 Hz and 74±12% at 20 Hz (all P<0.01), indicating they were predominantly neurogenic in nature. Neither the muscarinic antagonist atropine (10 µm), the adrenergic neurone blocker guanethidine (10 µm) nor desensitization of the purinergic receptors with α,ß-methylene ATP (10 µm) affected the contractile amplitude. Similarly, responses were not affected by the nitric oxide synthase inhibitor L-NNA (100 µm) or drugs that interfere with peptide neurotransmission (capsaicin, NK2 antagonist GR159897, protease inhibitors). In conclusion, electrical depolarization of the nerves present in the porcine urothelium/lamina propria results in frequency-dependent contractions, which are predominantly neurogenic in nature. These contractions are resistant to drugs that inhibit the adrenergic, cholinergic and purinergic systems. The neurotransmitter involved in the responses of this tissue is therefore unknown but does not appear to be a peptide.
Assuntos
Mucosa/inervação , Contração Muscular , Músculo Liso/inervação , Neurônios/metabolismo , Transmissão Sináptica , Bexiga Urinária/inervação , Urotélio/inervação , Matadouros , Acetilcolina/antagonistas & inibidores , Acetilcolina/fisiologia , Trifosfato de Adenosina/antagonistas & inibidores , Trifosfato de Adenosina/fisiologia , Animais , Estimulação Elétrica , Técnicas In Vitro , Mucosa/efeitos dos fármacos , Mucosa/metabolismo , Contração Muscular/efeitos dos fármacos , Músculo Liso/efeitos dos fármacos , Músculo Liso/metabolismo , Proteínas do Tecido Nervoso/antagonistas & inibidores , Proteínas do Tecido Nervoso/metabolismo , Neurônios/efeitos dos fármacos , Neurotransmissores/farmacologia , Óxido Nítrico/antagonistas & inibidores , Óxido Nítrico/fisiologia , Norepinefrina/antagonistas & inibidores , Norepinefrina/fisiologia , Parassimpatolíticos/farmacologia , Receptores de Neurotransmissores/antagonistas & inibidores , Receptores de Neurotransmissores/metabolismo , Sus scrofa , Transmissão Sináptica/efeitos dos fármacos , Tetrodotoxina/farmacologia , Bexiga Urinária/efeitos dos fármacos , Bexiga Urinária/metabolismo , Urotélio/efeitos dos fármacos , Urotélio/metabolismoRESUMO
Development of kinase-targeted therapies for central nervous system (CNS) diseases is a great challenge. Glycogen synthase kinase 3 (GSK-3) offers a great potential for severe CNS unmet diseases, being one of the inhibitors on clinical trials for different tauopathies. Following our hypothesis based on the enhanced reactivity of residue Cys199 in the binding site of GSK-3, we examine here the suitability of phenylhalomethylketones as irreversible inhibitors. Our data confirm that the halomethylketone unit is essential for the inhibitory activity. Moreover, addition of the halomethylketone moiety to reversible inhibitors turned them into irreversible inhibitors with IC(50) values in the nanomolar range. Overall, the results point out that these compounds might be useful pharmacological tools to explore physiological and pathological processes related to signaling pathways regulated by GSK-3 opening new avenues for the discovery of novel GSK-3 inhibitors.
Assuntos
Fármacos do Sistema Nervoso Central/síntese química , Quinase 3 da Glicogênio Sintase/antagonistas & inibidores , Cetonas/síntese química , Trifosfato de Adenosina/química , Animais , Sítios de Ligação , Bovinos , Fármacos do Sistema Nervoso Central/química , Fármacos do Sistema Nervoso Central/farmacologia , Cerebelo/citologia , Desenho de Fármacos , Humanos , Técnicas In Vitro , Cetonas/química , Cetonas/farmacologia , Camundongos , Modelos Moleculares , Doenças Neurodegenerativas/tratamento farmacológico , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Fosforilação , Ligação Proteica , Ratos , Receptores de Neurotransmissores/antagonistas & inibidores , Estereoisomerismo , Relação Estrutura-Atividade , Proteínas tau/metabolismoRESUMO
The requirement of c-Myb during erythropoiesis spurred an interest in identifying c-Myb target genes that are important for erythroid development. Here, we determined that the neuropeptide neuromedin U (NmU) is a c-Myb target gene. Silencing NmU, c-myb, or NmU's cognate receptor NMUR1 expression in human CD34(+) cells impaired burst-forming unit-erythroid (BFU-E) and colony-forming unit-erythroid (CFU-E) formation compared with control. Exogenous addition of NmU peptide to NmU or c-myb siRNA-treated CD34(+) cells rescued BFU-E and yielded a greater number of CFU-E than observed with control. No rescue of BFU-E and CFU-E growth was observed when NmU peptide was exogenously added to NMUR1 siRNA-treated cells compared with NMUR1 siRNA-treated cells cultured without NmU peptide. In K562 and CD34(+) cells, NmU activated protein kinase C-ßII, a factor associated with hematopoietic differentiation-proliferation. CD34(+) cells cultured under erythroid-inducing conditions, with NmU peptide and erythropoietin added at day 6, revealed an increase in endogenous NmU and c-myb gene expression at day 8 and a 16% expansion of early erythroblasts at day 10 compared to cultures without NmU peptide. Combined, these data strongly support that the c-Myb target gene NmU functions as a novel cofactor for erythropoiesis and expands early erythroblasts.
Assuntos
Eritroblastos/metabolismo , Células Precursoras Eritroides/metabolismo , Eritropoese/fisiologia , Neuropeptídeos/genética , Proteínas Proto-Oncogênicas c-myb/metabolismo , Receptores de Neurotransmissores/metabolismo , Western Blotting , Diferenciação Celular , Células Cultivadas , Imunoprecipitação da Cromatina , Ensaio de Unidades Formadoras de Colônias , Citometria de Fluxo , Imunofluorescência , Humanos , Luciferases/metabolismo , Neuropeptídeos/antagonistas & inibidores , Neuropeptídeos/metabolismo , Fragmentos de Peptídeos/farmacologia , Proteínas Proto-Oncogênicas c-myb/antagonistas & inibidores , Proteínas Proto-Oncogênicas c-myb/genética , RNA Mensageiro/genética , RNA Interferente Pequeno/genética , Receptores de Neurotransmissores/antagonistas & inibidores , Receptores de Neurotransmissores/genética , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Neuromedin U (NMU) plays very important roles in the central nervous system. However, to date, any role of NMU in hippocampal neurons and the relevant mechanisms still remain unknown. In the present study, we report that NMU selectively inhibits L-type high-voltage-gated Ca(2+) channels (HVGCC) in mouse hippocampal neurons, in which NMU type 1 receptor (NMUR1), but not NMUR2, is endogenously expressed. In wild type mice, NMU (0.1 microM) reversibly inhibited HVGCC barium currents (I(Ba)) by approximately 28%, while in NMUR1(-/-) mice NMU had no significant effects. Intracellular infusion of GDP-beta-S or a selective antibody raised against the G(o)alpha, as well as pretreatment of the neurons with pertussis toxin, blocked the inhibitory effects of NMU, indicating the involvement of G(o)-protein. This NMUR1-mediated effect did not display the characteristics of a direct interaction between G-protein betagamma subunit (G(betagamma)) and L-type HVGCC, but was abolished by dialyzing cells with QEHA peptide or an antibody to the G(beta). The classical and novel protein kinase C (PKC) antagonist calphostin C, as well as phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002, abolished NMU responses, whereas the classical PKC antagonist Gö6976 had no such effects. Cells dialyzed with a PKC epsilon isoform (PKCepsilon) specific inhibitor peptide, GAVSLLPT, abolished NMU responses. In contrast, in cells dialyzed with an inactive PKCepsilon control scramble peptide, LSGTLPAV, no significant effects were observed. In summary, these results suggest that NMU inhibits L-type HVGCC via activation of NMUR1 and downstream G(betagamma), PI3K, and a novel PKCepsilon signaling pathway.
Assuntos
Canais de Cálcio Tipo L/metabolismo , Neurônios/metabolismo , Fosfatidilinositol 3-Quinase/metabolismo , Proteína Quinase C-épsilon/metabolismo , Receptores de Neurotransmissores/metabolismo , Sequência de Aminoácidos , Animais , Células Cultivadas , Cromonas/farmacologia , Subunidades beta da Proteína de Ligação ao GTP/metabolismo , Hipocampo/citologia , Camundongos , Morfolinas/farmacologia , Naftalenos/metabolismo , Neurônios/enzimologia , Neurônios/fisiologia , Neuropeptídeos/genética , Neuropeptídeos/metabolismo , Peptídeos/metabolismo , Toxina Pertussis/farmacologia , Receptores de Neurotransmissores/antagonistas & inibidores , Receptores de Neurotransmissores/fisiologia , Transdução de SinaisAssuntos
Sistemas de Liberação de Medicamentos/métodos , Neurofarmacologia/métodos , Domínios e Motivos de Interação entre Proteínas/efeitos dos fármacos , Psicofarmacologia/métodos , Animais , Fatores de Transcrição de Zíper de Leucina Básica/antagonistas & inibidores , Neoplasias da Mama/tratamento farmacológico , Proteínas de Transporte/antagonistas & inibidores , Doenças do Sistema Nervoso Central/tratamento farmacológico , Proteínas de Grupos de Complementação da Anemia de Fanconi/antagonistas & inibidores , Feminino , Proteínas de Arcabouço Homer , Humanos , Neoplasias Ovarianas/tratamento farmacológico , Ratos , Receptores de Neurotransmissores/agonistas , Receptores de Neurotransmissores/antagonistas & inibidores , Proteínas Supressoras de Tumor/antagonistas & inibidores , Ubiquitina Tiolesterase/antagonistas & inibidoresRESUMO
Responses to psychostimulants vary with age, but the molecular etiologies of these differences are largely unknown. The goal of the present research was to identify age-specific behavioral and molecular adaptations to cocaine and to elucidate the mechanisms involved therein. Postweanling, periadolescent, and adult male CD-1 mice were exposed to cocaine (20 mg/kg) for 7 d. The rewarding effects of cocaine were assessed, as were the response to a Trk antagonist and the regulation of dopamine and cAMP-regulated phosphoprotein, 32 kDa (DARPP-32). Cocaine was rewarding in both periadolescent and adult mice using a conditioned place preference procedure. In contrast, postweanling mice failed to demonstrate significant cocaine-induced place preference. Because components of the neurotrophin system including brain-derived neurotrophic factor and TrkB are developmentally regulated, their role in the age-specific effects of cocaine was determined using the Trk receptor antagonist K252a. Postweanling mice that received K252a before daily cocaine showed a significant place preference to the cocaine-paired environment that was not seen in the absence of K252a. DARPP-32 protein levels were significantly upregulated in the lateral region of the caudate-putamen exclusively in postweanling mice after chronic cocaine. Daily pretreatment with K252a attenuated the induction of DARPP-32 in the postweanling striatum. These data indicate that Trk neurotransmission plays a role in age-specific behavioral and molecular responses to cocaine and concurrently modulates DARPP-32 levels.
Assuntos
Cocaína/farmacologia , Atividade Motora/fisiologia , Receptor trkB/química , Receptor trkB/fisiologia , Fatores Etários , Animais , Animais Recém-Nascidos , Carbazóis/farmacologia , Condicionamento Psicológico/efeitos dos fármacos , Condicionamento Psicológico/fisiologia , Fosfoproteína 32 Regulada por cAMP e Dopamina/fisiologia , Alcaloides Indólicos/farmacologia , Masculino , Camundongos , Atividade Motora/efeitos dos fármacos , Receptor trkB/antagonistas & inibidores , Receptores de Neurotransmissores/antagonistas & inibidores , Receptores de Neurotransmissores/química , Receptores de Neurotransmissores/fisiologiaRESUMO
Postoperative nausea and vomiting (PONV) is one of the most common and distressing complications following surgery, and understanding the mechanism(s) underlying PONV is essential to providing optimal prophylaxis and/or treatment of PONV. The knowledge base of PONV physiology has significantly expanded over the past decade. This article reviews the risk factors for the development of PONV and the mechanisms of action of pharmacological agents (including antagonists of serotonin 5-HT(3), dopaminergic D(2), histamine H(1), muscarinic cholinergic, opioid and neurokinin NK(1) receptors) for the management (i.e. prophylaxis and treatment) of PONV. NK(1) receptor antagonists, with their unique mechanism of action, are a particularly promising area of research as they appear to be efficacious in preventing PONV during both the early and the late postoperative periods. A successful PONV management strategy includes: (i) identifying patients at risk; (ii) keeping the baseline risk low; and (iii) using a combination of antiemetics acting on different receptors in moderate- to high-risk patients.
Assuntos
Tratamento Farmacológico/métodos , Náusea e Vômito Pós-Operatórios/prevenção & controle , Receptores de Neurotransmissores/antagonistas & inibidores , Animais , Humanos , Metanálise como Assunto , Modelos Biológicos , Náusea e Vômito Pós-Operatórios/fisiopatologiaRESUMO
OBJECTIVE: Among antipsychotics, clozapine ranks highest in terms of the risk for weight gain and developing diabetes. However, the mechanism by which clozapine induces weight gain and diabetes remains unclear. The aim of this study was to determine the mechanism of clozapine-induced weight gain and hyperglycemia, and to clarify whether clozapine-induced hyperglycemia results from impairment of the system regulating appetite. METHODS: Circulatory glucose, insulin, leptin and ghrelin levels were analyzed after acute administration of clozapine in rats. Clozapine (10 mg/kg) or a vehicle was injected intraperitoneally and blood samples were collected at 0, 15, 30, and 60 min after the injection. Clozapine (5, 10 or 20 mg/kg) or the vehicle was given, and blood samples were collected at 30 min after the injection. Since clozapine has receptor affinity for multiple neurotransmitters, selective antagonists of it, including dopamine, serotonin, alpha-adrenergic, muscarine and histamine were administered to clarify the pathway of clozapine-induced blood glucose and changes in plasma ghrelin. RESULTS: Clozapine administration increased the blood glucose level at all time points (p<0.05) compared to controls. Plasma ghrelin was elevated at 30 min (p=0.0124) and 60 min (p=0.00152). Blood glucose was increased in rats given 5 (p=0.0344), 10 (p<0.0001), or 20 mg/kg (p<0.0001) clozapine, while plasma ghrelin was increased in rats treated with 10 mg/kg (p=0.0009) or 20 mg/kg (p=0.0059) clozapine. Blood glucose was increased in rats treated with a selective alpha1-adrenergic receptor antagonist (p<0.0001), while plasma ghrelin was significantly increased in rats given a selective alpha1- (p=0.025) or alpha2-adrenergic receptor antagonist (p=0.0003). CONCLUSIONS: Clozapine impairs glucose metabolism and the appetite-regulation system. Clozapine increases blood glucose independent of insulin. The antagonistic action of alpha-adrenergic receptors is one of the mechanisms that induces both hyperglycemia and elevation of ghrelin.
Assuntos
Antipsicóticos/farmacologia , Glicemia/metabolismo , Clozapina/farmacologia , Hormônios Peptídicos/metabolismo , Antagonistas Adrenérgicos alfa/farmacologia , Animais , Relação Dose-Resposta a Droga , Grelina , Insulina/sangue , Leptina/sangue , Masculino , Ratos , Ratos Sprague-Dawley , Receptores de Neurotransmissores/antagonistas & inibidoresRESUMO
The brain controls fertility through release of gonadotropin-releasing hormone (GnRH), but the mechanisms underlying action potential patterning and GnRH release are not understood. We investigated whether GnRH neurons exhibit afterdepolarizing potentials (ADPs) and whether these are modified by reproductive state. Whole-cell current-clamp recordings of GnRH neurons in brain slices from ovariectomized mice revealed a slow ADP (sADP) after action potentials generated by brief current injection. Generating two or four spikes enhanced sADP amplitude and duration. sADP amplitude was not affected by blocking selected neurotransmitter/neuromodulator receptors, delayed-rectifier potassium channels, calcium-dependent cation channels, or hyperpolarization-activated cation channels but was halved by the calcium channel blocker cadmium and abolished by tetrodotoxin. Cadmium also reduced peak latency. Intrinsic mechanisms underlying the sADP were investigated using voltage-clamp protocols simulating action potential waveforms. A single action potential produced an inward current, which increased after double and quadruple stimulation. Cadmium did not affect current amplitude but reduced peak latency. Pretreatment with blockers of calcium-activated potassium currents (I(KCa)) reproduced this shift and blocked subsequent cadmium-induced changes, suggesting cadmium changes latency indirectly by blocking I(KCa). Tetrodotoxin abolished the inward current, suggesting that it is carried by sodium. In contrast, I(KCa) blockers increased the inward current, indicating that I(KCa) may oppose generation of the sADP. Strong sADPs were suprathreshold, generating repetitive spontaneous firing. I(ADP), sADP, and excitability were enhanced by in vivo estradiol, which triggers a preovulatory surge of GnRH release. Physiological feedback modification of this inward current and resulting sADP may modulate action potential firing and subsequent GnRH release.
Assuntos
Núcleo Hipotalâmico Anterior/metabolismo , Fertilidade/fisiologia , Hormônio Liberador de Gonadotropina/metabolismo , Neurônios/metabolismo , Área Pré-Óptica/metabolismo , Canais de Sódio/metabolismo , Potenciais de Ação/efeitos dos fármacos , Potenciais de Ação/fisiologia , Animais , Núcleo Hipotalâmico Anterior/citologia , Núcleo Hipotalâmico Anterior/efeitos dos fármacos , Bloqueadores dos Canais de Cálcio/farmacologia , Canais de Cálcio/efeitos dos fármacos , Canais de Cálcio/metabolismo , Estradiol/metabolismo , Ciclo Estral/efeitos dos fármacos , Ciclo Estral/fisiologia , Retroalimentação Fisiológica/fisiologia , Feminino , Fertilidade/efeitos dos fármacos , Camundongos , Neurônios/citologia , Neurônios/efeitos dos fármacos , Técnicas de Cultura de Órgãos , Ovariectomia , Técnicas de Patch-Clamp , Área Pré-Óptica/citologia , Área Pré-Óptica/efeitos dos fármacos , Tempo de Reação/efeitos dos fármacos , Tempo de Reação/fisiologia , Receptores de Neurotransmissores/antagonistas & inibidores , Receptores de Neurotransmissores/metabolismo , Bloqueadores dos Canais de Sódio/farmacologia , Canais de Sódio/efeitos dos fármacos , Tetrodotoxina/farmacologiaRESUMO
Chemical signal transduction from the cell surface to organelles was studied in unicellular vegetative (Equisetum arvense) and generative (Hippeastrum hybridum pollen) microspores of plants. Neurotransmitters acetylcholine, dopamine, and serotonin, their agonists and antagonists, Na+, K+, and Ca2+ channel blockers, as well as forskolin and theophylline (agents increasing the intracellular level of cyclic adenosine monophosphate) were used as chemical signals. Both types of microspores exposed to neurotransmitters, their agonists, forskolin, and theophylline demonstrated growth activation, while neurotransmitter antagonists and ion channel blockers inhibited this process. No stimulating effects of neurotransmitters were observed for cells pretreated with the antagonists and ion channel blockers. Pretreatment with ion channel blockers and then by anticontractile agents (cytochalasin B or colchicine) either had no effect or increased the inhibition of microspore growth. Pathways of chemical signal transduction from the cell surface to organelles are discussed.
Assuntos
Equisetum/fisiologia , Liliaceae/fisiologia , Colchicina/análogos & derivados , Colchicina/farmacologia , Colforsina/farmacologia , AMP Cíclico/metabolismo , Citocalasina B/farmacologia , Equisetum/efeitos dos fármacos , Equisetum/crescimento & desenvolvimento , Ativação do Canal Iônico , Canais Iônicos/efeitos dos fármacos , Liliaceae/efeitos dos fármacos , Liliaceae/crescimento & desenvolvimento , Neurotransmissores/agonistas , Neurotransmissores/antagonistas & inibidores , Neurotransmissores/farmacologia , Inibidores de Fosfodiesterase/farmacologia , Receptores de Neurotransmissores/agonistas , Receptores de Neurotransmissores/antagonistas & inibidores , Receptores de Neurotransmissores/metabolismo , Transdução de Sinais , Esporos/efeitos dos fármacos , Esporos/crescimento & desenvolvimento , Esporos/fisiologia , Teofilina/farmacologiaRESUMO
Many of the motoric features that define Parkinson's disease (PD) result primarily from the loss of dopaminergic neurons of the substantia nigra. l-dopa remains at present the most powerful symptomatic drug for the treatment of this condition. However, motor complications of chronic l-dopa treatment have emerged as a major limitation of this therapy. Slowing or delaying the progression of the disease with neuroprotective therapies may delay the need for l-dopa. In the past few years, novel insight into the pathogenetic mechanisms of neurodegeneration in PD has been provided. Mitochondrial function deficiency, increased oxidative stress, apoptosis, excitotoxicity, and inflammation are part of the processes that ultimately result in neurodegeneration. Drugs that are now under clinical scrutiny as neuroprotectant include molecules that combine one or more of the following properties: (1) monoamine oxidase inhibition (rasagiline, safinamide); (2) mitochondrial enhancement (coenzyme Q10, creatine); (3) antiapoptotic activity; (4) anti-inflammatory activity; (5) protein aggregation inhibition; (6) neurotrophic activity. In advanced Parkinson's disease, the combination of disease progression and l-dopa therapy leads to the development of motor response complications, particularly wearing off, on off, dyskinesias and dystonias. The nonphysiologic pulsatile stimulation of striatal dopamine receptors, produced by the currently available dopaminergic drugs, may trigger a dysregulation of many neurotransmitter systems within the basal ganglia, mainly localized on medium spiny striatal neurons. These include alterations of glutamatergic, serotonergic, adrenergic and adenosine A(2A) receptors. Novel strategies for pharmacological intervention with nondopaminergic treatments hold the promise of providing effective control or reversal of motor response complications. Of particular interest are NMDA and AMPA antagonists or drugs acting on 5-HT subtype 2A, alpha2-adrenergic, and adenosine A(2) receptors. Future strategies may also target pre- and postsynaptic components that regulate firing pattern of basal ganglia neurons, such as synaptic vesicle proteins, nonsynaptic gap junction communication mechanisms, or signal transduction systems that modulate the phosphorylation state of glutamatergic receptors.