Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 18 de 18
Filtrar
1.
J Allergy Clin Immunol ; 150(3): 690-700, 2022 09.
Artigo em Inglês | MEDLINE | ID: mdl-35240144

RESUMO

BACKGROUND: Atopic dermatitis is a chronic inflammatory skin disease with persistent and severe itch among its hallmark features. Significant increases in type 2 cytokines (ie, IL-4, IL-13, IL-31) have been documented in acute atopic dermatitis lesions and lead to multifaceted downstream effects, including inflammation, epidermal barrier dysfunction, and itch. OBJECTIVE: The primary objective of preclinical studies reported here was to test direct effects of IL-13 and an anti-IL-13 mAb, lebrikizumab, in a human dorsal root ganglion model in itch amplification, neuronal excitability, and transcriptional downstream targets. METHODS: Neuroactive effects were assessed via live cell calcium imaging, electric field stimulation, and RNA sequencing of human dorsal root ganglia stimulated with IL-13 alone or in combination with lebrikizumab. RESULTS: These preclinical findings suggest that IL-13 plays a direct enhancer role in multiple itch and neuroactive pathways as well as transcriptional downstream effects, and provide key insights into the mechanistic basis for lebrikizumab's anti-itch effects. CONCLUSION: IL-13 is a potent enhancer of neuronal responses to different itch stimuli, consistent with the neuroimmune axis contributing to chronic itch-associated inflammatory skin disease, and blockade of this cytokine pathway may provide a therapeutic approach.


Assuntos
Dermatite Atópica , Anticorpos Monoclonais/farmacologia , Anticorpos Monoclonais/uso terapêutico , Antipruriginosos/farmacologia , Citocinas/metabolismo , Humanos , Prurido , Pele
2.
Cereb Cortex ; 30(4): 2372-2388, 2020 04 14.
Artigo em Inglês | MEDLINE | ID: mdl-31761935

RESUMO

Huntington's disease (HD) is a neurodegenerative disorder characterized by involuntary movements, cognitive deficits, and psychiatric disturbances. Although evidence indicates that projections from motor cortical areas play a key role in the development of dysfunctional striatal activity and motor phenotype, little is known about the changes in cortical microcircuits and their role in the development of the HD phenotype. Here we used two-photon laser-scanning microscopy to evaluate network dynamics of motor cortical neurons in layers II/III in behaving transgenic R6/2 and knock-in Q175+/- mice. Symptomatic R6/2 mice displayed increased motion manifested by a significantly greater number of motion epochs, whereas symptomatic Q175 mice displayed decreased motion. In both models, calcium transients in symptomatic mice displayed reduced amplitude, suggesting decreased bursting activity. Changes in frequency were genotype- and time-dependent; for R6/2 mice, the frequency was reduced during both motion and nonmotion, whereas in symptomatic Q175 mice, the reduction only occurred during nonmotion. In presymptomatic Q175 mice, frequency was increased during both behavioral states. Interneuronal correlation coefficients were generally decreased in both models, suggesting disrupted interneuronal communication in HD cerebral cortex. These results indicate similar and contrasting effects of the HD mutation on cortical ensemble activity depending on mouse model and disease stage.


Assuntos
Cálcio , Modelos Animais de Doenças , Doença de Huntington/diagnóstico por imagem , Doença de Huntington/genética , Córtex Motor/diagnóstico por imagem , Rede Nervosa/diagnóstico por imagem , Animais , Cálcio/metabolismo , Feminino , Doença de Huntington/metabolismo , Masculino , Camundongos , Camundongos Transgênicos , Microscopia de Fluorescência por Excitação Multifotônica/métodos , Córtex Motor/metabolismo , Neurônios Motores/metabolismo , Rede Nervosa/metabolismo
3.
J Neurosci ; 36(1): 65-79, 2016 Jan 06.
Artigo em Inglês | MEDLINE | ID: mdl-26740650

RESUMO

Retrospective epidemiological studies show an inverse correlation between susceptibility to Parkinson's disease and a person's history of tobacco use. Animal model studies suggest nicotine as a neuroprotective agent and nicotinic acetylcholine (ACh) receptors (nAChRs) as targets for neuroprotection, but the underlying neuroprotective mechanism(s) are unknown. We cultured mouse ventral midbrain neurons for 3 weeks. Ten to 20% of neurons were dopaminergic (DA), revealed by tyrosine hydroxylase (TH) immunoreactivity. We evoked mild endoplasmic reticulum (ER) stress with tunicamycin (Tu), producing modest increases in the level of nuclear ATF6, phosphorylated eukaryotic initiation factor 2α, nuclear XBP1, and the downstream proapoptotic effector nuclear C/EBP homologous protein. We incubated cultures for 2 weeks with 200 nm nicotine, the approximate steady-state concentration between cigarette smoking or vaping, or during nicotine patch use. Nicotine incubation suppressed Tu-induced ER stress and the unfolded protein response (UPR). Study of mice with fluorescent nAChR subunits showed that the cultured TH+ neurons displayed α4, α6, and ß3 nAChR subunit expression and ACh-evoked currents. Gene expression profile in cultures from TH-eGFP mice showed that the TH+ neurons also express several other genes associated with DA release. Nicotine also upregulated ACh-induced currents in DA neurons by ∼2.5-fold. Thus, nicotine, at a concentration too low to activate an appreciable fraction of plasma membrane nAChRs, induces two sequelae of pharmacological chaperoning in the ER: UPR suppression and nAChR upregulation. Therefore, one mechanism of neuroprotection by nicotine is pharmacological chaperoning, leading to UPR suppression. Measuring this pathway may help in assessing neuroprotection. SIGNIFICANCE STATEMENT: Parkinson's disease (PD) cannot yet be cured or prevented. However, many retrospective epidemiological studies reveal that PD is diagnosed less frequently in tobacco users. Existing programs attempting to develop nicotinic drugs that might exert this apparent neuroprotective effect are asking whether agonists, antagonists, partial agonists, or channel blockers show the most promise. The underlying logic resembles the previous development of varenicline for smoking cessation. We studied whether, and how, nicotine produces neuroprotective effects in cultured dopaminergic neurons, an experimentally tractable, mechanistically revealing neuronal system. We show that nicotine, operating via nicotinic receptors, does protect these neurons against endoplasmic reticulum stress. However, the mechanism is probably "inside-out": pharmacological chaperoning in the endoplasmic reticulum. This cellular-level insight could help to guide neuroprotective strategies.


Assuntos
Potenciais de Ação/fisiologia , Neurônios Dopaminérgicos/fisiologia , Nicotiana/química , Nicotina/administração & dosagem , Fumaça , Resposta a Proteínas não Dobradas/fisiologia , Potenciais de Ação/efeitos dos fármacos , Animais , Células Cultivadas , Neurônios Dopaminérgicos/efeitos dos fármacos , Relação Dose-Resposta a Droga , Feminino , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Fármacos Neuroprotetores/administração & dosagem , Resposta a Proteínas não Dobradas/efeitos dos fármacos
4.
J Biol Chem ; 289(38): 26451-26463, 2014 Sep 19.
Artigo em Inglês | MEDLINE | ID: mdl-25056953

RESUMO

The α7 nicotinic acetylcholine receptor gene (CHRNA7) is linked to schizophrenia. A partial duplication of CHRNA7 (CHRFAM7A) is found in humans on 15q13-14. Exon 6 of CHRFAM7A harbors a 2-bp deletion polymorphism, CHRFAM7AΔ2bp, which is also associated with schizophrenia. To understand the effects of the duplicated subunits on α7 receptors, we fused α7, dupα7, and dupΔα7 subunits with various fluorescent proteins. The duplicated subunits co-localized with full-length α7 subunits in mouse neuroblastoma cells (Neuro2a) as well as rat hippocampal neurons. We investigated the interaction between the duplicated subunits and full-length α7 by measuring Förster resonance energy transfer using donor recovery after photobleaching and fluorescence lifetime imaging microscopy. The results revealed that the duplicated proteins co-assemble with α7. In electrophysiological studies, Leu at the 9'-position in the M2 membrane-spanning segment was replaced with Cys in dupα7 or dupΔα7, and constructs were co-transfected with full-length α7 in Neuro2a cells. Exposure to ethylammonium methanethiosulfonate inhibited acetylcholine-induced currents, showing that the assembled functional nicotinic acetylcholine receptors (nAChRs) included the duplicated subunit. Incorporation of dupα7 and dupΔα7 subunits modestly changes the sensitivity of receptors to choline and varenicline. Thus, the duplicated proteins are assembled and transported to the cell membrane together with full-length α7 subunits and alter the function of the nAChRs. The characterization of dupα7 and dupΔα7 as well as their influence on α7 nAChRs may help explain the pathophysiology of schizophrenia and may suggest therapeutic strategies.


Assuntos
Receptor Nicotínico de Acetilcolina alfa7/metabolismo , Acetilcolina/farmacologia , Animais , Benzazepinas/farmacologia , Linhagem Celular Tumoral , Colina/farmacologia , Duplicação Gênica , Humanos , Potenciais da Membrana/efeitos dos fármacos , Camundongos , Agonistas Nicotínicos/farmacologia , Multimerização Proteica , Estrutura Quaternária de Proteína , Subunidades Proteicas/genética , Subunidades Proteicas/metabolismo , Transporte Proteico , Quinoxalinas/farmacologia , Ratos , Esquizofrenia/genética , Deleção de Sequência , Vareniclina , Receptor Nicotínico de Acetilcolina alfa7/genética
5.
J Neurophysiol ; 113(7): 2953-66, 2015 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-25673747

RESUMO

The Q175 knockin mouse model of Huntington's disease (HD) carries a CAG trinucleotide expansion of the human mutant huntingtin allele in its native mouse genomic context and recapitulates the genotype more closely than transgenic models. In this study we examined the progression of changes in intrinsic membrane properties and excitatory and inhibitory synaptic transmission, using whole cell patch-clamp recordings of medium-sized spiny neurons (MSNs) in the dorsolateral striatum and cortical pyramidal neurons (CPNs) in layers 2/3 of the primary motor cortex in brain slices from heterozygous (Q175(+/-)) and homozygous (Q175(+/+)) mice. Input resistance in MSNs from Q175(+/+) and Q175(+/-) mice was significantly increased compared with wild-type (WT) littermates beginning at 2 mo. Furthermore, the frequency of spontaneous and miniature excitatory postsynaptic currents (EPSCs) was significantly reduced in MSNs from Q175(+/+) and Q175(+/-) mice compared with WTs beginning at 7 mo. In contrast, the frequency of spontaneous inhibitory postsynaptic currents (sIPSCs) and IPSC-to-EPSC ratios were increased in MSNs from Q175(+/+) mice beginning at 2 mo. Morphologically, significant decreases in spine density of MSNs from Q175(+/-) and Q175(+/+) mice occurred at 7 and 12 mo. In CPNs, sIPSC frequencies and IPSC-to-EPSC ratios were significantly increased in Q175(+/-) mice compared with WTs at 12 mo. There were no changes in intrinsic membrane properties or morphology. In summary, we show a number of alterations in electrophysiological and morphological properties of MSNs in Q175 mice that are similar to other HD mouse models. However, unlike other models, CPN inhibitory activity is increased in Q175(+/-) mice, indicating reduced cortical excitability.


Assuntos
Corpo Estriado/fisiopatologia , Modelos Animais de Doenças , Potenciais Pós-Sinápticos Excitadores , Doença de Huntington/fisiopatologia , Inibição Neural , Células Piramidais , Animais , Tamanho Celular , Corpo Estriado/patologia , Feminino , Técnicas de Introdução de Genes , Masculino , Camundongos , Plasticidade Neuronal
6.
J Neurosci ; 32(40): 13987-99, 2012 Oct 03.
Artigo em Inglês | MEDLINE | ID: mdl-23035106

RESUMO

Traf2 and NcK interacting kinase (TNiK) contains serine-threonine kinase and scaffold domains and has been implicated in cell proliferation and glutamate receptor regulation in vitro. Here we report its role in vivo using mice carrying a knock-out mutation. TNiK binds protein complexes in the synapse linking it to the NMDA receptor (NMDAR) via AKAP9. NMDAR and metabotropic receptors bidirectionally regulate TNiK phosphorylation and TNiK is required for AMPA expression and synaptic function. TNiK also organizes nuclear complexes and in the absence of TNiK, there was a marked elevation in GSK3ß and phosphorylation levels of its cognate phosphorylation sites on NeuroD1 with alterations in Wnt pathway signaling. We observed impairments in dentate gyrus neurogenesis in TNiK knock-out mice and cognitive testing using the touchscreen apparatus revealed impairments in pattern separation on a test of spatial discrimination. Object-location paired associate learning, which is dependent on glutamatergic signaling, was also impaired. Additionally, TNiK knock-out mice displayed hyperlocomotor behavior that could be rapidly reversed by GSK3ß inhibitors, indicating the potential for pharmacological rescue of a behavioral phenotype. These data establish TNiK as a critical regulator of cognitive functions and suggest it may play a regulatory role in diseases impacting on its interacting proteins and complexes.


Assuntos
Aprendizagem por Associação/fisiologia , Transtornos Cognitivos/enzimologia , Giro Denteado/enzimologia , Aprendizagem por Discriminação/fisiologia , Proteínas do Tecido Nervoso/fisiologia , Densidade Pós-Sináptica/enzimologia , Proteínas Serina-Treonina Quinases/fisiologia , Detecção de Sinal Psicológico/fisiologia , Percepção Espacial/fisiologia , Animais , Núcleo Celular/enzimologia , Transtornos Cognitivos/fisiopatologia , Giro Denteado/patologia , Ácido Glutâmico/fisiologia , Quinase 3 da Glicogênio Sintase/antagonistas & inibidores , Quinase 3 da Glicogênio Sintase/fisiologia , Glicogênio Sintase Quinase 3 beta , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Potenciais Pós-Sinápticos em Miniatura/fisiologia , Proteínas do Tecido Nervoso/deficiência , Neurogênese/fisiologia , Fenótipo , Fosforilação , Densidade Pós-Sináptica/fisiologia , Processamento de Proteína Pós-Traducional , Proteínas Serina-Treonina Quinases/biossíntese , Proteínas Serina-Treonina Quinases/deficiência , Proteínas Serina-Treonina Quinases/genética , Proteínas Recombinantes de Fusão/fisiologia
7.
J Neurosci ; 31(45): 16194-207, 2011 Nov 09.
Artigo em Inglês | MEDLINE | ID: mdl-22072671

RESUMO

Densin is an abundant scaffold protein in the postsynaptic density (PSD) that forms a high-affinity complex with αCaMKII and α-actinin. To assess the function of densin, we created a mouse line with a null mutation in the gene encoding it (LRRC7). Homozygous knock-out mice display a wide variety of abnormal behaviors that are often considered endophenotypes of schizophrenia and autism spectrum disorders. At the cellular level, loss of densin results in reduced levels of α-actinin in the brain and selective reduction in the localization of mGluR5 and DISC1 in the PSD fraction, whereas the amounts of ionotropic glutamate receptors and other prominent PSD proteins are unchanged. In addition, deletion of densin results in impairment of mGluR- and NMDA receptor-dependent forms of long-term depression, alters the early dynamics of regulation of CaMKII by NMDA-type glutamate receptors, and produces a change in spine morphology. These results indicate that densin influences the function of mGluRs and CaMKII at synapses and contributes to localization of mGluR5 and DISC1 in the PSD fraction. They are consistent with the hypothesis that mutations that disrupt the organization and/or dynamics of postsynaptic signaling complexes in excitatory synapses can cause behavioral endophenotypes of mental illness.


Assuntos
Regulação da Expressão Gênica/genética , Transtornos Mentais/genética , Proteínas do Tecido Nervoso/metabolismo , Densidade Pós-Sináptica/metabolismo , Receptores de Ácido Caínico/metabolismo , Sialoglicoproteínas/deficiência , Actinas/metabolismo , Agressão/fisiologia , Animais , Comportamento Animal/fisiologia , Bicuculina/farmacologia , Peso Corporal/genética , Proteína Quinase Tipo 2 Dependente de Cálcio-Calmodulina/metabolismo , Células Cultivadas , Espinhas Dendríticas/metabolismo , Modelos Animais de Doenças , Embrião de Mamíferos , Endofenótipos , Comportamento Exploratório/fisiologia , Feminino , Antagonistas GABAérgicos/farmacologia , Antagonistas de Receptores de GABA-A/farmacologia , Genótipo , Proteína Glial Fibrilar Ácida/metabolismo , Glicina/farmacologia , Proteínas de Fluorescência Verde/genética , Hipocampo/citologia , Técnicas In Vitro , Inibição Psicológica , Potenciação de Longa Duração/genética , Depressão Sináptica de Longo Prazo/genética , Masculino , Potenciais da Membrana/efeitos dos fármacos , Potenciais da Membrana/genética , Memória de Curto Prazo/fisiologia , Transtornos Mentais/patologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Atividade Motora/genética , Força Muscular/genética , Mutação/genética , Proteínas do Tecido Nervoso/genética , Neurônios/citologia , Neurônios/efeitos dos fármacos , Técnicas de Patch-Clamp , Picrotoxina/farmacologia , Desempenho Psicomotor/fisiologia , Receptores de AMPA/genética , Receptores de Ácido Caínico/genética , Reconhecimento Psicológico/fisiologia , Teste de Desempenho do Rota-Rod , Estatísticas não Paramétricas , Fatores de Tempo
8.
Learn Mem ; 18(2): 118-27, 2011 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-21257779

RESUMO

Through protein interactions mediated by their cytoplasmic C termini the GluN2A and GluN2B subunits of NMDA receptors (NMDARs) have a key role in the formation of NMDAR signaling complexes at excitatory synapses. Although these signaling complexes are thought to have a crucial role in NMDAR-dependent forms of synaptic plasticity such as long-term potentiation (LTP), the role of the C terminus of GluN2A in coupling NMDARs to LTP enhancing and/or suppressing signaling pathways is unclear. To address this issue we examined the induction of LTP in the hippocampal CA1 region in mice lacking the C terminus of endogenous GluN2A subunits (GluN2AΔC/ΔC). Our results show that truncation of GluN2A subunits produces robust, but highly frequency-dependent, deficits in LTP and a reduction in basal levels of extracellular signal regulated kinase 2 (ERK2) activation and phosphorylation of AMPA receptor GluA1 subunits at a protein kinase A site (serine 845). Consistent with the notion that these signaling deficits contribute to the deficits in LTP in GluN2AΔC/ΔC mice, activating ERK2 and increasing GluA1 S845 phosphorylation through activation of ß-adrenergic receptors rescued the induction of LTP in these mutants. Together, our results indicate that the capacity of excitatory synapses to undergo plasticity in response to different patterns of activity is dependent on the coupling of specific signaling pathways to the intracellular domains of the NMDARs and that abnormal plasticity resulting from mutations in NMDARs can be reduced by activation of key neuromodulatory transmitter receptors that engage converging signaling pathways.


Assuntos
Hipocampo/fisiologia , Potenciação de Longa Duração/genética , Mutação/genética , Receptores Adrenérgicos beta/metabolismo , Agonistas Adrenérgicos beta/farmacologia , Animais , Fenômenos Biofísicos/genética , Estimulação Elétrica/métodos , Agonistas de Aminoácidos Excitatórios/farmacologia , Hipocampo/citologia , Hipocampo/efeitos dos fármacos , Técnicas In Vitro , Isoproterenol/farmacologia , Potenciação de Longa Duração/efeitos dos fármacos , Potenciação de Longa Duração/fisiologia , Camundongos , Proteína Quinase 1 Ativada por Mitógeno/metabolismo , N-Metilaspartato/farmacologia , Técnicas de Patch-Clamp/métodos , Fragmentos de Peptídeos/metabolismo , Fosforilação/efeitos dos fármacos , Células Piramidais/efeitos dos fármacos , Células Piramidais/fisiologia , Receptores de N-Metil-D-Aspartato/deficiência , Receptores de N-Metil-D-Aspartato/genética , Serina/metabolismo
9.
Sci Rep ; 11(1): 12014, 2021 06 08.
Artigo em Inglês | MEDLINE | ID: mdl-34103608

RESUMO

Late sodium current (late INa) inhibition has been proposed to suppress the incidence of arrhythmias generated by pathological states or induced by drugs. However, the role of late INa in the human heart is still poorly understood. We therefore investigated the role of this conductance in arrhythmias using adult primary cardiomyocytes and tissues from donor hearts. Potentiation of late INa with ATX-II (anemonia sulcata toxin II) and E-4031 (selective blocker of the hERG channel) slowed the kinetics of action potential repolarization, impaired Ca2+ homeostasis, increased contractility, and increased the manifestation of arrhythmia markers. These effects could be reversed by late INa inhibitors, ranolazine and GS-967. We also report that atrial tissues from donor hearts affected by atrial fibrillation exhibit arrhythmia markers in the absence of drug treatment and inhibition of late INa with GS-967 leads to a significant reduction in arrhythmic behaviour. These findings reveal a critical role for the late INa in cardiac arrhythmias and suggest that inhibition of this conductance could provide an effective therapeutic strategy. Finally, this study highlights the utility of human ex-vivo heart models for advancing cardiac translational sciences.


Assuntos
Fibrilação Atrial/metabolismo , Canal de Potássio ERG1/metabolismo , Potenciais da Membrana , Modelos Cardiovasculares , Miócitos Cardíacos/metabolismo , Adulto , Cálcio/metabolismo , Venenos de Cnidários/farmacologia , Canal de Potássio ERG1/antagonistas & inibidores , Átrios do Coração/metabolismo , Humanos , Miócitos Cardíacos/patologia , Piperidinas/farmacologia , Piridinas/farmacologia , Ranolazina/farmacologia , Sódio , Triazóis/farmacologia
10.
Sci Rep ; 10(1): 7692, 2020 05 06.
Artigo em Inglês | MEDLINE | ID: mdl-32376974

RESUMO

Effects of non-cardiac drugs on cardiac contractility can lead to serious adverse events. Furthermore, programs aimed at treating heart failure have had limited success and this therapeutic area remains a major unmet medical need. The challenges in assessing drug effect on cardiac contractility point to the fundamental translational value of the current preclinical models. Therefore, we sought to develop an adult human primary cardiomyocyte contractility model that has the potential to provide a predictive preclinical approach for simultaneously predicting drug-induced inotropic effect (sarcomere shortening) and generating multi-parameter data to profile different mechanisms of action based on cluster analysis of a set of 12 contractility parameters. We report that 17 positive and 9 negative inotropes covering diverse mechanisms of action exerted concentration-dependent increases and decreases in sarcomere shortening, respectively. Interestingly, the multiparametric readout allowed for the differentiation of inotropes operating via distinct mechanisms. Hierarchical clustering of contractility transient parameters, coupled with principal component analysis, enabled the classification of subsets of both positive as well as negative inotropes, in a mechanism-related mode. Thus, human cardiomyocyte contractility model could accurately facilitate informed mechanistic-based decision making, risk management and discovery of molecules with the most desirable pharmacological profile for the correction of heart failure.


Assuntos
Cardiotônicos/farmacologia , Contração Miocárdica/efeitos dos fármacos , Miócitos Cardíacos/efeitos dos fármacos , Sarcômeros/efeitos dos fármacos , Adulto , Diferenciação Celular/efeitos dos fármacos , Análise por Conglomerados , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Adulto Jovem
11.
Front Neurosci ; 13: 176, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30881283

RESUMO

The hippocampus, a structure essential for spatial navigation and memory undergoes anatomical and functional changes during chronic stress. Here, we investigate the effects of chronic stress on the ability of place cells to encode the neural representation of a linear track. To model physiological conditions of chronic stress on hippocampal function, transgenic mice expressing the genetically encoded calcium indicator GCaMP6f in CA1 pyramidal neurons were chronically administered with 40 µg/ml of cortisol for 8 weeks. Cortisol-treated mice exhibited symptoms typically observed during chronic stress, including diminished reward seeking behavior and reduced adrenal gland and spleen weights. In vivo imaging of hippocampal cellular activity during linear track running behavior revealed a reduced number of cells that could be recruited to encode spatial position, despite an unchanged overall number of active cells, in cortisol-treated mice. The properties of the remaining place cells that could be recruited to encode spatial information, however, was unperturbed. Bayesian decoders trained to estimate the mouse's position on the track using single neuron activity data demonstrated reduced performance in a cue richness-dependent fashion in cortisol-treated animals. The performance of decoders utilizing data from the entire neuronal ensemble was unaffected by cortisol treatment. Finally, to test the hypothesis that an antidepressant drug could prevent the effects of cortisol, we orally administered a group of mice with 10 mg/kg citalopram during cortisol administration. Citalopram prevented the cortisol-induced decrease in single-neuron decoder performance but failed to significantly prevent anhedonia and the cortisol-induced reduction in the proportion place cells. The dysfunction observed at the single-neuron level indicates that chronic stress may impair the ability of the hippocampus to encode individual neural representations of the mouse's spatial position, a function pivotal to forming an accurate navigational map of the mouse's external environment; however, the hippocampal ensemble as a whole is resilient to any cortisol-induced insults to single neuronal place cell function on the linear track.

12.
Schizophr Res ; 99(1-3): 286-93, 2008 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-18248794

RESUMO

OBJECTIVE: Earlier studies described gaze discrimination impairment in schizophrenia. The purpose of this study was to compare gaze discrimination abilities and associated brain activation in persons with stable schizophrenia and matched controls. METHODS: 13 schizophrenia and 12 healthy participants underwent a gaze discrimination task with face stimuli rotated at 0, 4 and 8 degrees deviation. During fMRI with BOLD imaging, subjects were asked to identify whether a face was making eye contact. Subject-level parameter estimates for BOLD signal change were entered into an orientation by group mixed effect repeated measures ANOVA. RESULTS: Gaze discrimination performance did not differ between groups. Patients showed decreased activation in areas of bilateral inferior frontal and occipital areas, and select temporo-limbic regions, including amygdala. Groups differed by activation patterns according to gaze deviation. In controls, faces with 4 degrees deviation produced higher activation in frontal and temporal regions. In patients, 0 degrees deviation produced increased activation in amygdala and areas of temporal neocortex. CONCLUSIONS: Despite similar gaze discrimination abilities, schizophrenia patients exhibit decreased brain activation in areas associated with executive, emotional and visual processing. Controls exhibited increased activation associated with the more difficult task in select frontal and temporal regions. Patients exhibited increased activation associated with direct gaze in temporal regions, which may relate to common symptoms.


Assuntos
Atenção/fisiologia , Córtex Cerebral/fisiopatologia , Discriminação Psicológica/fisiologia , Face , Fixação Ocular/fisiologia , Processamento de Imagem Assistida por Computador , Imageamento por Ressonância Magnética , Orientação/fisiologia , Oxigênio/sangue , Reconhecimento Visual de Modelos/fisiologia , Esquizofrenia/fisiopatologia , Psicologia do Esquizofrênico , Adulto , Tonsila do Cerebelo/fisiopatologia , Mapeamento Encefálico , Dominância Cerebral/fisiologia , Feminino , Lobo Frontal/fisiopatologia , Humanos , Masculino , Pessoa de Meia-Idade , Rede Nervosa/fisiopatologia , Esquizofrenia/diagnóstico , Lobo Temporal/fisiopatologia
13.
Curr Protoc Pharmacol ; 82(1): e42, 2018 09.
Artigo em Inglês | MEDLINE | ID: mdl-30129249

RESUMO

Imaging neuronal activity in awake behaving mice with miniature fluorescence microscopes requires the implementation of a variety of procedures. Surgeries are performed to gain access to the cell population of interest and to implant microscope components. After a recovery period, mice are trained to exhibit a desired behavior. Finally, neuronal activity is imaged and synchronized with that behavior. To take full advantage of the technology, selection of the calcium indicator and experimental design must be carefully considered. In this article, we explain the procedures and considerations that are critical for obtaining high-quality calcium imaging data. As an example, we describe how to utilize miniature fluorescence microscopy to image hippocampal place cell activity during linear track running in Thy1.GCaMP6f transgenic mice. © 2018 by John Wiley & Sons, Inc.


Assuntos
Células de Lugar/fisiologia , Animais , Comportamento Animal , Cálcio/fisiologia , Hipocampo/citologia , Hipocampo/fisiologia , Camundongos Transgênicos , Microscopia de Fluorescência , Atividade Motora
14.
Cell Rep ; 25(4): 841-851.e4, 2018 10 23.
Artigo em Inglês | MEDLINE | ID: mdl-30355491

RESUMO

The GluN2 subtype (2A versus 2B) determines biophysical properties and signaling of forebrain NMDA receptors (NMDARs). During development, GluN2A becomes incorporated into previously GluN2B-dominated NMDARs. This "switch" is proposed to be driven by distinct features of GluN2 cytoplasmic C-terminal domains (CTDs), including a unique CaMKII interaction site in GluN2B that drives removal from the synapse. However, these models remain untested in the context of endogenous NMDARs. We show that, although mutating the endogenous GluN2B CaMKII site has secondary effects on GluN2B CTD phosphorylation, the developmental changes in NMDAR composition occur normally and measures of plasticity and synaptogenesis are unaffected. Moreover, the switch proceeds normally in mice that have the GluN2A CTD replaced by that of GluN2B and commences without an observable decline in GluN2B levels but is impaired by GluN2A haploinsufficiency. Thus, GluN2A expression levels, and not GluN2 subtype-specific CTD-driven events, are the overriding factor in the developmental switch in NMDAR composition.


Assuntos
Receptores de N-Metil-D-Aspartato/química , Receptores de N-Metil-D-Aspartato/metabolismo , Sequência de Aminoácidos , Animais , Sítios de Ligação , Proteína Quinase Tipo 2 Dependente de Cálcio-Calmodulina/metabolismo , Potenciação de Longa Duração , Camundongos Endogâmicos C57BL , Mutação/genética , Neurogênese , Fosforilação , Subunidades Proteicas/metabolismo , Ratos , Receptores de N-Metil-D-Aspartato/genética , Sinapses/metabolismo , Ritmo Teta/fisiologia
15.
Schizophr Res ; 94(1-3): 253-63, 2007 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-17583481

RESUMO

OBJECTIVE: Schizophrenia patients demonstrate impaired emotional processing that may be due, in part, to impaired facial emotion recognition. This study examined event-related potential (ERP) responses to emotional faces in schizophrenia patients and controls to determine when, in the temporal processing stream, patient abnormalities occur. METHOD: 16 patients and 16 healthy control participants performed a facial emotion recognition task. Very sad, somewhat sad, neutral, somewhat happy, and very happy faces were each presented for 100 ms. Subjects indicated whether each face was "Happy", "Neutral", or "Sad". Evoked potential data were obtained using a 32-channel EEG system. RESULTS: Controls performed better than patients in recognizing facial emotions. In patients, better recognition of happy faces correlated with less severe negative symptoms. Four ERP components corresponding to the P100, N170, N250, and P300 were identified. Group differences were noted for the N170 "face processing" component that underlies the structural encoding of facial features, but not for the subsequent N250 "affect modulation" component. Higher amplitude of the N170 response to sad faces was correlated with less severe delusional symptoms. Although P300 abnormalities were found, the variance of this component was explained by the earlier N170 response. CONCLUSION: Patients with schizophrenia demonstrate abnormalities in early visual encoding of facial features that precedes the ERP response typically associated with facial affect recognition. This suggests that affect recognition deficits, at least for happy and sad discrimination, are secondary to faulty structural encoding of faces. The association of abnormal face encoding with delusions may denote the physiological basis for clinical misidentification syndromes.


Assuntos
Transtornos Cognitivos/etiologia , Emoções Manifestas , Expressão Facial , Reconhecimento Psicológico , Esquizofrenia/complicações , Percepção Social , Adulto , Encéfalo/fisiopatologia , Transtornos Cognitivos/diagnóstico , Eletroencefalografia , Feminino , Humanos , Masculino , Esquizofrenia/fisiopatologia , Percepção Visual/fisiologia
16.
J Neurosci ; 23(9): 3820-5, 2003 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-12736352

RESUMO

Since the discovery of facial asymmetries in emotional expressions of humans and other primates, hypotheses have related the greater left-hemiface intensity to right-hemispheric dominance in emotion processing. However, the difficulty of creating true frontal views of facial expressions in two-dimensional photographs has confounded efforts to better understand the phenomenon. We have recently described a method for obtaining three-dimensional photographs of posed and evoked emotional expressions and used these stimuli to investigate both intensity of expression and accuracy of recognizing emotion in chimeric faces constructed from only left- or right-side composites. The participant population included 38 (19 male, 19 female) African-American, Caucasian, and Asian adults. They were presented with chimeric composites generated from faces of eight actors and eight actresses showing four emotions: happiness, sadness, anger, and fear, each in posed and evoked conditions. We replicated the finding that emotions are expressed more intensely in the left hemiface for all emotions and conditions, with the exception of evoked anger, which was expressed more intensely in the right hemiface. In contrast, the results indicated that emotional expressions are recognized more efficiently in the right hemiface, indicating that the right hemiface expresses emotions more accurately. The double dissociation between the laterality of expression intensity and that of recognition efficiency supports the notion that the two kinds of processes may have distinct neural substrates. Evoked anger is uniquely expressed more intensely and accurately on the side of the face that projects to the viewer's right hemisphere, dominant in emotion recognition.


Assuntos
Dominância Cerebral/fisiologia , Emoções/fisiologia , Face , Reconhecimento Visual de Modelos/fisiologia , Adolescente , Adulto , Ira , Medo , Feminino , Lateralidade Funcional/fisiologia , Pesar , Felicidade , Humanos , Processamento de Imagem Assistida por Computador/métodos , Masculino , Estimulação Luminosa/métodos , Grupos Raciais
17.
J Neurosci Methods ; 141(1): 61-73, 2005 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-15585289

RESUMO

We present a novel methodology for quantitative analysis of changes in facial display as the intensity of an emotion evolves from neutral to peak expression. The face is modeled as a combination of regions and their boundaries. An expression change in a face is characterized and quantified through a combination of non-rigid (elastic) deformations, i.e., expansions and contractions of these facial regions. After elastic interpolation, this yields a geometry-based high-dimensional 2D shape transformation, which is used to register regions defined on subjects (i.e., faces with expression) to those defined on the reference template face (a neutral face). This shape transformation produces a vector-valued deformation field and is used to define a scalar valued regional volumetric difference (RVD) function, which characterizes and quantifies the facial expression. The approach is applied to a standardized database consisting of single images of professional actors expressing emotions at predefined intensities. We perform a detailed analysis of the deformations generated and the regional volumetric differences computed for expressions. We were able to quantify subtle changes in expression that can distinguish the intended emotions. A model for the average expression of specific emotions was also constructed using the RVD maps. This method can be applied in basic and clinical investigations of facial affect and its neural substrates.


Assuntos
Algoritmos , Expressão Facial , Processamento de Imagem Assistida por Computador/métodos , Fotografação/métodos , Adolescente , Adulto , Idoso , Emoções/fisiologia , Feminino , Humanos , Processamento de Imagem Assistida por Computador/instrumentação , Masculino , Pessoa de Meia-Idade , Comportamento Social
18.
Nat Neurosci ; 16(1): 25-32, 2013 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-23201971

RESUMO

Two genome duplications early in the vertebrate lineage expanded gene families, including GluN2 subunits of the NMDA receptor. Diversification between the four mammalian GluN2 proteins occurred primarily at their intracellular C-terminal domains (CTDs). To identify shared ancestral functions and diversified subunit-specific functions, we exchanged the exons encoding the GluN2A (also known as Grin2a) and GluN2B (also known as Grin2b) CTDs in two knock-in mice and analyzed the mice's biochemistry, synaptic physiology, and multiple learned and innate behaviors. The eight behaviors were genetically separated into four groups, including one group comprising three types of learning linked to conserved GluN2A/B regions. In contrast, the remaining five behaviors exhibited subunit-specific regulation. GluN2A/B CTD diversification conferred differential binding to cytoplasmic MAGUK proteins and differential forms of long-term potentiation. These data indicate that vertebrate behavior and synaptic signaling acquired increased complexity from the duplication and diversification of ancestral GluN2 genes.


Assuntos
Comportamento Animal/fisiologia , Citoplasma/metabolismo , Evolução Molecular , Plasticidade Neuronal/fisiologia , Receptores de N-Metil-D-Aspartato/metabolismo , Sinapses/fisiologia , Animais , Biofísica , Estimulação Elétrica , Embrião de Mamíferos , Células-Tronco Embrionárias , Emoções/fisiologia , Potenciais Pós-Sinápticos Excitadores/efeitos dos fármacos , Potenciais Pós-Sinápticos Excitadores/genética , Humanos , Imunoprecipitação , Ativação do Canal Iônico/efeitos dos fármacos , Ativação do Canal Iônico/genética , Aprendizagem/fisiologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Motivação/genética , Atividade Motora/genética , Mutação/genética , Plasticidade Neuronal/efeitos dos fármacos , Plasticidade Neuronal/genética , Técnicas de Patch-Clamp , Fenótipo , Estrutura Terciária de Proteína/genética , Receptores de N-Metil-D-Aspartato/genética , Sinapses/efeitos dos fármacos , Sinapses/genética
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA