Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 35
Filtrar
Mais filtros










Base de dados
Intervalo de ano de publicação
1.
Cells ; 13(12)2024 Jun 18.
Artigo em Inglês | MEDLINE | ID: mdl-38920682

RESUMO

Neuroplasticity in the amygdala and its central nucleus (CeA) is linked to pain modulation and pain behaviors, but cellular mechanisms are not well understood. Here, we addressed the role of small-conductance Ca2+-activated potassium (SK) channels in pain-related amygdala plasticity. The facilitatory effects of the intra-CeA application of an SK channel blocker (apamin) on the pain behaviors of control rats were lost in a neuropathic pain model, whereas an SK channel activator (NS309) inhibited pain behaviors in neuropathic rats but not in sham controls, suggesting the loss of the inhibitory behavioral effects of amygdala SK channels. Brain slice electrophysiology found hyperexcitability of CeA neurons in the neuropathic pain condition due to the loss of SK channel-mediated medium afterhyperpolarization (mAHP), which was accompanied by decreased SK2 channel protein and mRNA expression, consistent with a pretranscriptional mechanisms. The underlying mechanisms involved the epigenetic silencing of the SK2 gene due to the increased DNA methylation of the CpG island of the SK2 promoter region and the change in methylated CpG sites in the CeA in neuropathic pain. This study identified the epigenetic dysregulation of SK channels in the amygdala (CeA) as a novel mechanism of neuropathic pain-related plasticity and behavior that could be targeted to control abnormally enhanced amygdala activity and chronic neuropathic pain.


Assuntos
Tonsila do Cerebelo , Epigênese Genética , Neuralgia , Canais de Potássio Ativados por Cálcio de Condutância Baixa , Animais , Masculino , Ratos , Tonsila do Cerebelo/metabolismo , Tonsila do Cerebelo/fisiopatologia , Comportamento Animal/efeitos dos fármacos , Metilação de DNA/genética , Neuralgia/metabolismo , Neuralgia/genética , Neuralgia/fisiopatologia , Neurônios/metabolismo , Ratos Sprague-Dawley , Canais de Potássio Ativados por Cálcio de Condutância Baixa/metabolismo , Canais de Potássio Ativados por Cálcio de Condutância Baixa/genética
2.
Front Pharmacol ; 15: 1368634, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38576475

RESUMO

Introduction: Pain is a clinically relevant health care issue with limited therapeutic options, creating the need for new and improved analgesic strategies. The amygdala is a limbic brain region critically involved in the regulation of emotional-affective components of pain and in pain modulation. The central nucleus of amygdala (CeA) serves major output functions and receives nociceptive information via the external lateral parabrachial nucleus (PB). While amygdala neuroplasticity has been linked causally to pain behaviors, non-neuronal pain mechanisms in this region remain to be explored. As an essential part of the neuroimmune system, astrocytes that represent about 40-50% of glia cells within the central nervous system, are required for physiological neuronal functions, but their role in the amygdala remains to be determined for pain conditions. In this study, we measured time-specific astrocyte activation in the CeA in a neuropathic pain model (spinal nerve ligation, SNL) and assessed the effects of astrocyte inhibition on amygdala neuroplasticity and pain-like behaviors in the pain condition. Methods and Results: Glial fibrillary acidic protein (GFAP, astrocytic marker) immunoreactivity and mRNA expression were increased at the chronic (4 weeks post-SNL), but not acute (1 week post-SNL), stage of neuropathic pain. In order to determine the contribution of astrocytes to amygdala pain-mechanisms, we used fluorocitric acid (FCA), a selective inhibitor of astrocyte metabolism. Whole-cell patch-clamp recordings were performed from neurons in the laterocapsular division of the CeA (CeLC) obtained from chronic neuropathic rats. Pre-incubation of brain slices with FCA (100 µM, 1 h), increased excitability through altered hyperpolarization-activated current (Ih) functions, without significantly affecting synaptic responses at the PB-CeLC synapse. Intra-CeA injection of FCA (100 µM) had facilitatory effects on mechanical withdrawal thresholds (von Frey and paw pressure tests) and emotional-affective behaviors (evoked vocalizations), but not on facial grimace score and anxiety-like behaviors (open field test), in chronic neuropathic rats. Selective inhibition of astrocytes by FCA was confirmed with immunohistochemical analyses showing decreased astrocytic GFAP, but not NeuN, signal in the CeA. Discussion: Overall, these results suggest a complex modulation of amygdala pain functions by astrocytes and provide evidence for beneficial functions of astrocytes in CeA in chronic neuropathic pain.

3.
Neurobiol Dis ; 190: 106361, 2024 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-37992784

RESUMO

The prefrontal cortex is a crucial regulator of alcohol drinking, and dependence, and other behavioral phenotypes associated with AUD. Comprehensive identification of cell-type specific transcriptomic changes in alcohol dependence will improve our understanding of mechanisms underlying the excessive alcohol use associated with alcohol dependence and will refine targets for therapeutic development. We performed single nucleus RNA sequencing (snRNA-seq) and Visium spatial gene expression profiling on the medial prefrontal cortex (mPFC) obtained from C57BL/6 J mice exposed to the two-bottle choice-chronic intermittent ethanol (CIE) vapor exposure (2BC-CIE, defined as dependent group) paradigm which models phenotypes of alcohol dependence including escalation of alcohol drinking. Gene co-expression network analysis and differential expression analysis identified highly dysregulated co-expression networks in multiple cell types. Dysregulated modules and their hub genes suggest novel understudied targets for studying molecular mechanisms contributing to the alcohol dependence state. A subtype of inhibitory neurons was the most alcohol-sensitive cell type and contained a downregulated gene co-expression module; the hub gene for this module is Cpa6, a gene previously identified by GWAS to be associated with excessive alcohol consumption. We identified an astrocytic Gpc5 module significantly upregulated in the alcohol-dependent group. To our knowledge, there are no studies linking Cpa6 and Gpc5 to the alcohol-dependent phenotype. We also identified neuroinflammation related gene expression changes in multiple cell types, specifically enriched in microglia, further implicating neuroinflammation in the escalation of alcohol drinking. Here, we present a comprehensive atlas of cell-type specific alcohol dependence mediated gene expression changes in the mPFC and identify novel cell type-specific targets implicated in alcohol dependence.


Assuntos
Alcoolismo , Animais , Camundongos , Alcoolismo/genética , Doenças Neuroinflamatórias , Camundongos Endogâmicos C57BL , Encéfalo/metabolismo , Córtex Pré-Frontal/metabolismo , Etanol/toxicidade
4.
Neuropharmacology ; 242: 109768, 2024 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-37865137

RESUMO

Stress increases alcohol consumption in dependent animals and contributes to the development of alcohol use disorder. The nucleus of the solitary tract (NTS) is a critical brainstem region for integrating and relaying central and peripheral signals to regulate stress responses, but it is not known if it plays a role in alcohol dependence- or in stress-induced escalations in alcohol drinking in dependent mice. Here, we used RNA-sequencing and bioinformatics analyses to study molecular adaptations in the NTS of C57BL/6J male mice that underwent an ethanol drinking procedure that uses exposure to chronic intermittent ethanol (CIE) vapor, forced swim stress (FSS), or both conditions (CIE + FSS). Transcriptome profiling was performed at three different times after the last vapor cycle (0-hr, 72-hr, and 168-hr) to identify changes in gene expression associated with different stages of ethanol intoxication and withdrawal. In the CIE and CIE + FSS groups at 0-hr, there was upregulation of genes enriched for cellular response to type I interferon (IFN) and type I IFN- and cytokine-mediated signaling pathways, while the FSS group showed upregulation of neuronal genes. IFN signaling was the top gene network positively correlated with ethanol consumption levels in the CIE and CIE + FSS groups. Results from different analyses (differential gene expression, weighted gene coexpression network analysis, and rank-rank hypergeometric overlap) indicated that activation of type I IFN signaling would be expected to increase ethanol consumption. The CIE and CIE + FSS groups also shared an immune signature in the NTS as has been demonstrated in other brain regions after chronic ethanol exposure. A temporal-based clustering analysis revealed a unique expression pattern in the CIE + FSS group that suggests the interaction of these two stressors produces adaptations in synaptic and glial functions that may drive stress-induced drinking.


Assuntos
Alcoolismo , Masculino , Animais , Camundongos , Alcoolismo/genética , Transcriptoma , Núcleo Solitário , Camundongos Endogâmicos C57BL , Etanol/farmacologia , Consumo de Bebidas Alcoólicas/genética
5.
Curr Top Behav Neurosci ; 64: 289-305, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37584834

RESUMO

Extinction is the process by which the memory of a learned conditioned association decreases over time and with introduction of new associations. It is a vital part of fear learning, and it is critical to recovery in multiple fear-related disorders, including Specific and Social Phobias, Panic Disorder, Obsessive Compulsive Disorder (OCD), and Posttraumatic Stress Disorder (PTSD). The process of extinction is also the underlying mechanism for recovery in gold-standard therapies for PTSD, including prolonged exposure, cognitive processing therapy, eye movement desensitization and procession, as well as other empirically-based paradigms. Pharmacological modulators of extinction are thus promising targets for treatment of fear-related disorders. We focus here on emerging psychopharmacological treatments to facilitate extinction: D-cycloserine, scopolamine, losartan, ketamine, and 3,4-methylenedioxymethamphetamine. We also provide an overview of recent advances in molecular pathways that show promise as targets for extincion and inhibitory learning, including pathways related to cannabinoid, brain-derived neurotrophic factor, hypothalamic-pituitary-adrenal signaling, and promising work in neurosteroid compounds.


Assuntos
Transtorno Obsessivo-Compulsivo , Transtornos de Estresse Pós-Traumáticos , Humanos , Medo , Extinção Psicológica , Transtornos de Estresse Pós-Traumáticos/tratamento farmacológico , Aprendizagem
6.
Int J Mol Sci ; 24(15)2023 Jul 26.
Artigo em Inglês | MEDLINE | ID: mdl-37569320

RESUMO

Chronic pain presents a therapeutic challenge due to the highly complex interplay of sensory, emotional-affective and cognitive factors. The mechanisms of the transition from acute to chronic pain are not well understood. We hypothesized that neuroimmune mechanisms in the amygdala, a brain region involved in the emotional-affective component of pain and pain modulation, play an important role through high motility group box 1 (Hmgb1), a pro-inflammatory molecule that has been linked to neuroimmune signaling in spinal nociception. Transcriptomic analysis revealed an upregulation of Hmgb1 mRNA in the right but not left central nucleus of the amygdala (CeA) at the chronic stage of a spinal nerve ligation (SNL) rat model of neuropathic pain. Hmgb1 silencing with a stereotaxic injection of siRNA for Hmgb1 into the right CeA of adult male and female rats 1 week after (post-treatment), but not 2 weeks before (pre-treatment) SNL induction decreased mechanical hypersensitivity and emotional-affective responses, but not anxiety-like behaviors, measured 4 weeks after SNL. Immunohistochemical data suggest that neurons are a major source of Hmgb1 in the CeA. Therefore, Hmgb1 in the amygdala may contribute to the transition from acute to chronic neuropathic pain, and the inhibition of Hmgb1 at a subacute time point can mitigate neuropathic pain.


Assuntos
Dor Crônica , Neuralgia , Animais , Feminino , Masculino , Ratos , Tonsila do Cerebelo , Neuralgia/genética , Neuralgia/terapia , Neurônios/fisiologia , Ratos Sprague-Dawley
7.
Radiat Oncol J ; 41(4): 267-273, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-38185931

RESUMO

PURPOSE: We evaluate various approaches to target volume definition and boost delivery in patients with complete response to neoadjuvant systemic therapy (NST) who were treated by radiotherapy without a surgery. MATERIALS AND METHODS: A pathological complete response (pCR) was diagnosed in 21 of 27 patients included in "surgery de-escalation" prospective observation study. Clips were placed in the primary tumor volume (PrTV) before NST and during the vacuum aspiration biopsy. Twenty patients with pCR underwent the whole breast irradiation and a boost to the PrTV. High-dose rate brachytherapy (HDRB) was the basic technique for boost delivery. Finally, we identified the value of fused images (computed tomography [CT] before NST with simulation CT), clips and their combination for an accurate boost delivery. RESULTS: A complete overlap between PrTV on pre-treatment CT with the localization of the clips on simulation CT was mentioned in 10, partial mismatch in three patients. In 12 of these 13 women, HDRB was successfully used for the boost delivery. In five cases we mentioned a marked discrepancy between the PrTV on fused images and the topography of the clips. In other two women we did not find clips on simulation CT. The fused images in five of these seven patients showed anatomical landmarks (scar, fibrosis) used for identification of the gross tumor volume. In all 20 women with pCR (average follow-up of 16.6 months), there were no locoregional recurrences. CONCLUSION: Combination of the clips with fusion of pre-NST and simulation CTs is important for an accurate boost delivery.

8.
Front Pharmacol ; 13: 903978, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35694266

RESUMO

Functional pain syndromes (FPS) occur in the absence of identifiable tissue injury or noxious events and include conditions such as migraine, fibromyalgia, and others. Stressors are very common triggers of pain attacks in various FPS conditions. It has been recently demonstrated that kappa opioid receptors (KOR) in the central nucleus of amygdala (CeA) contribute to FPS conditions, but underlying mechanisms remain unclear. The CeA is rich in KOR and encompasses major output pathways involving extra-amygdalar projections of corticotropin releasing factor (CRF) expressing neurons. Here we tested the hypothesis that KOR blockade in the CeA in a rat model of FPS reduces pain-like and nocifensive behaviors by restoring inhibition of CeA-CRF neurons. Intra-CeA administration of a KOR antagonist (nor-BNI) decreased mechanical hypersensitivity and affective and anxiety-like behaviors in a stress-induced FPS model. In systems electrophysiology experiments in anesthetized rats, intra-CeA application of nor-BNI reduced spontaneous firing and responsiveness of CeA neurons to peripheral stimulation. In brain slice whole-cell patch-clamp recordings, nor-BNI increased feedforward inhibitory transmission evoked by optogenetic and electrical stimulation of parabrachial afferents, but had no effect on monosynaptic excitatory transmission. Nor-BNI decreased frequency, but not amplitude, of spontaneous inhibitory synaptic currents, suggesting a presynaptic action. Blocking KOR receptors in stress-induced FPS conditions may therefore represent a novel therapeutic strategy.

9.
Neurobiol Stress ; 14: 100330, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33997155

RESUMO

Post-traumatic stress disorder (PTSD) is a devastating illness with treatment that is effective in only approximately half of the population. This limited rate of response highlights the necessity for research into underlying individual biological mechanisms that mediate development and progression of this disease, allowing for identification of patient-specific treatments. PTSD has clear sex differences in both risk and symptom patterns. Thus, one approach is to characterize trauma-related changes between men and women who exhibit differences in treatment efficacy and response to trauma. Recent technological advances in sequencing have identified several genomic loci and transcriptional changes that are associated with post-trauma symptomatology. However, although the diagnosis of PTSD is more prevalent in women, the genetic factors underlying sex differences remain poorly understood. Here, we review recent work that highlights current understanding and limitations in the field of sex differences in PTSD and related symptomatology.

10.
Ann Clin Psychiatry ; 33(2): 116-123, 2021 05.
Artigo em Inglês | MEDLINE | ID: mdl-33878286

RESUMO

BACKGROUND: Clozapine is an effective antipsychotic for treatment-resistant schizophrenia. One limitation of clozapine use is required monitoring of absolute neutrophil count (ANC) because of the risk of clozapine-induced neutropenia. Standard monitoring requires venous blood draws, which is a significant barrier to clozapine use. METHODS: This study assesses the feasibility of use and physician and patient satisfaction of a novel point-of-care (POC) measure of ANC using Athelas One, a device that calculates white blood cell count and ANC using a fingerstick blood sample. This is a subanalysis of a prospective, open-label clinical trial of clozapine treatment, during which patients received a venous blood draw and a capillary fingerstick at baseline and Week 2 of the study, and completed a 5-point Likert scale, comparing the 2 methods. RESULTS: Patients reported benefits from the fingerstick technology, including POC testing being important for their doctors and their health, improved treatment, avoiding sending blood away, and convenience. There was a trend for less concern about the effects of blood draws on health with a fingerstick, and greater physician satisfaction with POC sampling. CONCLUSIONS: This study suggests the feasibility, satisfaction, and ease by both clinicians and patients of using POC testing for ANC monitoring during clozapine treatment.


Assuntos
Antipsicóticos , Clozapina , Antipsicóticos/efeitos adversos , Clozapina/efeitos adversos , Estudos de Viabilidade , Humanos , Contagem de Leucócitos , Neutrófilos , Medidas de Resultados Relatados pelo Paciente , Satisfação do Paciente , Satisfação Pessoal , Sistemas Automatizados de Assistência Junto ao Leito , Estudos Prospectivos
11.
Brain Sci ; 10(5)2020 May 02.
Artigo em Inglês | MEDLINE | ID: mdl-32370184

RESUMO

Chronic stress is a known contributing factor to the development of drug and alcohol addiction. Animal models have previously shown that repeated forced swim stress promotes escalated alcohol consumption in dependent animals. To investigate the underlying molecular adaptations associated with stress and chronic alcohol exposure, RNA-sequencing and bioinformatics analyses were conducted on the prefrontal cortex (CTX) of male C57BL/6J mice that were behaviorally tested for either non-dependent alcohol consumption (CTL), chronic intermittent ethanol (CIE) vapor dependent alcohol consumption, repeated bouts of forced swim stress alone (FSS), and chronic intermittent ethanol with forced swim stress (CIE + FSS). Brain tissue from each group was collected at 0-h, 72-h, and 168-h following the final test to determine long-lasting molecular changes associated with maladaptive behavior. Our results demonstrate unique temporal patterns and persistent changes in coordinately regulated gene expression systems with respect to the tested behavioral group. For example, increased expression of genes involved in "transmitter-gated ion channel activity" was only determined for CIE + FSS. Overall, our results provide a summary of transcriptomic adaptations across time within the CTX that are relevant to understanding the neurobiology of chronic alcohol exposure and stress.

12.
Emerg Infect Dis ; 25(12): 2264-2265, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31742511

RESUMO

We assessed IgM detection in Zika patients from the 2016 outbreak in Miami-Dade County, Florida, USA. Of those with positive or equivocal IgM after 12-19 months, 87% (26/30) had IgM 6 months later. In a survival analysis, ≈76% had IgM at 25 months. Zika virus IgM persists for years, complicating serologic diagnosis.


Assuntos
Anticorpos Antivirais/imunologia , Imunoglobulina M/imunologia , Infecção por Zika virus/epidemiologia , Infecção por Zika virus/imunologia , Zika virus/imunologia , Adulto , Idoso , Anticorpos Antivirais/sangue , Surtos de Doenças , Feminino , Florida/epidemiologia , Humanos , Imunoglobulina M/sangue , Masculino , Pessoa de Meia-Idade , Fatores de Tempo , Adulto Jovem , Zika virus/genética , Infecção por Zika virus/diagnóstico , Infecção por Zika virus/virologia
13.
Emerg Infect Dis ; 25(2): 299-303, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30666931

RESUMO

Data on the duration of detectable Zika virus-specific IgM in infected persons are limited. Neutralizing antibody cross-reactivity occurs between Zika virus and related flaviviruses, but the degree to which this confounds diagnosis is uncertain. We tested serum specimens collected 12-19 months after illness onset from patients with confirmed Zika virus disease for Zika virus IgM and Zika virus and dengue virus neutralizing antibodies. Among 62 participants, 45 (73%) had detectable Zika virus IgM and 12 (19%) had an equivocal result. Although all patients tested had Zika virus neutralizing antibodies, 39 (63%) also had neutralizing antibodies against dengue virus; of those, 12 (19%) had <4-fold difference between Zika virus and dengue virus titers, and 5 (8%) had dengue virus titer >4-fold higher than Zika virus titer. Prolonged detection of IgM and neutralizing antibody cross-reactivity make it difficult to determine the timing of Zika virus infection and differentiate between related flaviviruses.


Assuntos
Anticorpos Neutralizantes/imunologia , Anticorpos Antivirais/imunologia , Imunoglobulina M/imunologia , Infecção por Zika virus/imunologia , Infecção por Zika virus/virologia , Zika virus/imunologia , Adolescente , Adulto , Idoso , Anticorpos Neutralizantes/sangue , Anticorpos Antivirais/sangue , Criança , Reações Cruzadas/imunologia , Feminino , Humanos , Imunoglobulina M/sangue , Masculino , Pessoa de Meia-Idade , Gravidez , Fatores de Tempo , Adulto Jovem , Zika virus/genética
14.
Alcohol Clin Exp Res ; 42(5): 926-938, 2018 05.
Artigo em Inglês | MEDLINE | ID: mdl-29469962

RESUMO

BACKGROUND: Phosphodiesterase type 4 (PDE4) inhibitors produce widespread anti-inflammatory effects and reduce ethanol (EtOH) consumption in several rodent models. These drugs are potential treatments for several diseases, including central nervous system disorders, but clinical use is limited by their emetic activity. Apremilast is a selective PDE4 inhibitor with fewer gastrointestinal side effects that is FDA-approved for the treatment of psoriasis. METHODS: We measured the acute and chronic effects of apremilast on EtOH consumption in male and female C57BL/6J mice using the continuous and intermittent 24-hour 2-bottle choice drinking models. We also studied the effects of apremilast on preference for sucrose or saccharin, spontaneous locomotor activity, and blood EtOH clearance. Finally, apremilast levels in plasma, liver, and brain were measured 1 or 2 hours after injection. RESULTS: In the continuous and intermittent drinking tests, apremilast (15 to 50 mg/kg, p.o.) dose dependently reduced EtOH intake and preference in male and female mice. Higher doses of apremilast (30 to 50 mg/kg) also reduced total fluid intake in these mice. Chronic administration of apremilast (20 mg/kg) produced a stable reduction in EtOH consumption in both drinking tests with no effect on total fluid intake. The drinking effects were reversible after drug treatment was replaced with vehicle administration (saline) for 2 to 4 days. Six daily apremilast injections did not alter preference for saccharin or sucrose in male or female mice. Apremilast (20 mg/kg) transiently decreased spontaneous locomotor activity and did not alter blood EtOH clearance. The highest levels of apremilast were found in liver followed by plasma and brain. CONCLUSIONS: Apremilast produced stable reductions in voluntary EtOH consumption and was rapidly distributed to plasma and tissues (including the brain), suggesting that it may be an improved PDE4 inhibitor for medication development and repurposing efforts to treat alcohol abuse.


Assuntos
Consumo de Bebidas Alcoólicas/prevenção & controle , Preferências Alimentares/efeitos dos fármacos , Talidomida/análogos & derivados , Animais , Encéfalo/metabolismo , Comportamento de Escolha/efeitos dos fármacos , Etanol/sangue , Etanol/farmacocinética , Feminino , Fígado/metabolismo , Masculino , Camundongos , Atividade Motora/efeitos dos fármacos , Sacarina/farmacologia , Sacarose/farmacologia , Talidomida/sangue , Talidomida/farmacocinética , Talidomida/farmacologia
15.
eNeuro ; 3(5)2016.
Artigo em Inglês | MEDLINE | ID: mdl-27822501

RESUMO

Proinflammatory pathways in neuronal and non-neuronal cells are implicated in the acute and chronic effects of alcohol exposure in animal models and humans. The nuclear factor-κB (NF-κB) family of DNA transcription factors plays important roles in inflammatory diseases. The kinase IKKß mediates the phosphorylation and subsequent proteasomal degradation of cytosolic protein inhibitors of NF-κB, leading to activation of NF-κB. The role of IKKß as a potential regulator of excessive alcohol drinking had not previously been investigated. Based on previous findings that the overactivation of innate immune/inflammatory signaling promotes ethanol consumption, we hypothesized that inhibiting IKKß would limit/decrease drinking by preventing the activation of NF-κB. We studied the systemic effects of two pharmacological inhibitors of IKKß, TPCA-1 and sulfasalazine, on ethanol intake using continuous- and limited-access, two-bottle choice drinking tests in C57BL/6J mice. In both tests, TPCA-1 and sulfasalazine reduced ethanol intake and preference without changing total fluid intake or sweet taste preference. A virus expressing Cre recombinase was injected into the nucleus accumbens and central amygdala to selectively knock down IKKß in mice genetically engineered with a conditional Ikkb deletion (IkkbF/F ). Although IKKß was inhibited to some extent in astrocytes and microglia, neurons were a primary cellular target. Deletion of IKKß in either brain region reduced ethanol intake and preference in the continuous access two-bottle choice test without altering the preference for sucrose. Pharmacological and genetic inhibition of IKKß decreased voluntary ethanol consumption, providing initial support for IKKß as a potential therapeutic target for alcohol abuse.


Assuntos
Consumo de Bebidas Alcoólicas/metabolismo , Tonsila do Cerebelo/metabolismo , Quinase I-kappa B/antagonistas & inibidores , Núcleo Accumbens/metabolismo , Dissuasores de Álcool/farmacologia , Consumo de Bebidas Alcoólicas/patologia , Consumo de Bebidas Alcoólicas/terapia , Tonsila do Cerebelo/efeitos dos fármacos , Tonsila do Cerebelo/patologia , Animais , Depressores do Sistema Nervoso Central/administração & dosagem , Modelos Animais de Doenças , Etanol/administração & dosagem , Preferências Alimentares/efeitos dos fármacos , Preferências Alimentares/fisiologia , Técnicas de Silenciamento de Genes , Quinase I-kappa B/metabolismo , Masculino , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , NF-kappa B/metabolismo , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Neurônios/patologia , Núcleo Accumbens/efeitos dos fármacos , Núcleo Accumbens/patologia , RNA Mensageiro/metabolismo , Sacarina
16.
Sci Rep ; 6: 27618, 2016 06 10.
Artigo em Inglês | MEDLINE | ID: mdl-27283430

RESUMO

Peroxisome proliferator-activated receptors (PPARs) are nuclear hormone receptors that act as ligand-activated transcription factors. PPAR agonists have well-documented anti-inflammatory and neuroprotective roles in the central nervous system. Recent evidence suggests that PPAR agonists are attractive therapeutic agents for treating neurodegenerative diseases as well as addiction. However, the distribution of PPAR mRNA and protein in brain regions associated with these conditions (i.e. prefrontal cortex, nucleus accumbens, amygdala, ventral tegmental area) is not well defined. Moreover, the cell type specificity of PPARs in mouse and human brain tissue has yet to be investigated. We utilized quantitative PCR and double immunofluorescence microscopy to determine that both PPAR mRNA and protein are expressed ubiquitously throughout the adult mouse brain. We found that PPARs have unique cell type specificities that are consistent between species. PPARα was the only isotype to colocalize with all cell types in both adult mouse and adult human brain tissue. Overall, we observed a strong neuronal signature, which raises the possibility that PPAR agonists may be targeting neurons rather than glia to produce neuroprotection. Our results fill critical gaps in PPAR distribution and define novel cell type specificity profiles in the adult mouse and human brain.


Assuntos
Doenças Neurodegenerativas/genética , Receptores Ativados por Proliferador de Peroxissomo/genética , Isoformas de Proteínas/genética , Tonsila do Cerebelo/efeitos dos fármacos , Animais , Comunicação Celular/genética , Linhagem da Célula/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Isotipos de Imunoglobulinas , Camundongos , Doenças Neurodegenerativas/tratamento farmacológico , Doenças Neurodegenerativas/patologia , Neurônios/efeitos dos fármacos , Neurônios/patologia , Receptores Ativados por Proliferador de Peroxissomo/agonistas , Receptores Ativados por Proliferador de Peroxissomo/classificação , Córtex Pré-Frontal/efeitos dos fármacos , Córtex Pré-Frontal/patologia , Isoformas de Proteínas/classificação , RNA Mensageiro/efeitos dos fármacos
17.
Biochim Biophys Acta ; 1858(9): 2199-2207, 2016 09.
Artigo em Inglês | MEDLINE | ID: mdl-27342373

RESUMO

Reaction mixture for PGHS (prostaglandin-H-synthase) is a two-phase system including micellar hydrophobic phase and hydrophilic aqueous phase. Reagents added to the mixture are distributed between phases, thus concentrations of reagents dissolved in phases can differ significantly from their overall contents. Using dynamic light scattering we found that the hydrophobic phase produced by tween-20 consists of micelles, which radius (4-5nm) does not depend on either tween-20 overall content (0.1%-1% v/v) or arachidonic acid (AA) addition (10-1000µM) or PGHS addition (1µM). Tween-20 overall content changing from 0.1% to 2% v/v dramatically affected COX kinetic, but accounting AA distribution between phases allowed us to estimate "true" parameters, independent of the tween-20 overall content and the concentration of another substrate: KM(Ox) equals 9.8µM O2 in the aqueous phase or 0.0074bar in the gaseous phase, KM(AA) equals 5400µM AA in the phase of tween-20 micelles and 5400/PµM AA in the aqueous phase (P is the distribution ratio for the AA between the aqueous phase and the hydrophobic phase (P≫1000)). This approach allowed to evaluate PS, the distribution ratio for the AA between the hydrophobic phase and the PGHS active center (PS ~310). This coefficient indicates the AA selectivity toward the cyclooxygenase active center.


Assuntos
Ácido Araquidônico/química , Micelas , Polissorbatos/química , Prostaglandina-Endoperóxido Sintases/química , Animais , Interações Hidrofóbicas e Hidrofílicas , Luz , Masculino , Ratos , Espalhamento de Radiação
18.
Neural Dev ; 11: 2, 2016 Jan 20.
Artigo em Inglês | MEDLINE | ID: mdl-26791407

RESUMO

BACKGROUND: Charcot-Marie-Tooth2b (CMT2b) is an axonal form of a human neurodegenerative disease that preferentially affects sensory neurons. CMT2b is dominantly inherited and is characterized by unusually early onset, presenting in the second or third decade of life. Five missense mutations in the gene encoding Rab7 GTPase have been identified as causative in human CMT2b disease. Although several studies have modeled CMT2b disease in cultured neurons and in Drosophila, the mechanisms by which defective Rab7 leads to disease remain poorly understood. RESULTS: We used zebrafish to investigate the effects of CMT2b-associated Rab7 mutations in a vertebrate model. We generated transgenic animals expressing the CMT2b-associated mutant forms of Rab7 in sensory neurons, and show that these Rab7 variants cause neurodevelopmental defects, including defects in sensory axon growth, branching and pathfinding at early developmental stages. We also find reduced axon growth and branching in neurons expressing a constitutively active form of Rab7, suggesting these defects may be caused by Rab7 gain-of-function. Further, we use high-speed, high-resolution imaging of endosome transport in vivo and find that CMT2b-associated Rab7 variants cause reduced vesicle speeds, suggesting altered transport may underlie axon development defects. CONCLUSIONS: Our data provide new insight into how disease-associated alterations in Rab7 protein disrupt cellular function in vertebrate sensory neurons. Moreover, our findings suggest that defects in axon development may be a previously unrecognized component of CMT2b disease.


Assuntos
Axônios/fisiologia , Doença de Charcot-Marie-Tooth/embriologia , Doença de Charcot-Marie-Tooth/genética , Células Receptoras Sensoriais/fisiologia , Proteínas de Peixe-Zebra/genética , Proteínas rab de Ligação ao GTP/genética , Sequência de Aminoácidos , Animais , Animais Geneticamente Modificados , Axônios/patologia , Morte Celular , Doença de Charcot-Marie-Tooth/patologia , Modelos Animais de Doenças , Endossomos/genética , Endossomos/fisiologia , Laminopatias , Dados de Sequência Molecular , Mutação , Células Receptoras Sensoriais/metabolismo , Células Receptoras Sensoriais/patologia , Alinhamento de Sequência , Peixe-Zebra , Proteínas de Peixe-Zebra/química , Proteínas de Peixe-Zebra/metabolismo , Proteínas rab de Ligação ao GTP/química , Proteínas rab de Ligação ao GTP/metabolismo , proteínas de unión al GTP Rab7
19.
Ann Nucl Med ; 29(2): 177-83, 2015 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-25398309

RESUMO

OBJECTIVE: To determine diagnostic accuracy of SPECT, CT and SPECT-CT in axillary lymph node (LN) staging in breast cancer (BC). METHODS: Sixty consecutive patients with primary operable T1-3NxM0 BC were included in this study. All patients underwent SPECT-CT examination on Symbia-T16 scanner which consists of dual-head gamma camera combined with 16 slices diagnostic CT. SPECT-CT acquisition started 10-15 min after i/v injection of 740-1,000 MBq of 99mTc-MIBI. On CT images of axillary LN we analyzed following diagnostic signs: size (short axis more or less than 10 mm), shape (round or oval), cortical thickness and fat content (solid or with fat gate). Intensity of tracer uptake in axillary LN was classified as follows: grade (Gr) I-background, Gr II-slightly above background, Gr III-intense but below uptake in muscles, Gr IV-as high as in muscles. Histological examination of dissected LN was used as gold standard. RESULTS: Various combinations of CT signs of axillary LN involvement demonstrated moderate diagnostic value with best results characterized by low (55 %) sensitivity (SEN), 97 % specificity (SP) and 83 % accuracy (AC). Intensive (Gr IV) uptake of 99mTc-MIBI in axillary LN characterized by low (55 %) SEN, high (100 %) SP and moderate (84 %) AC. Combination of CT and SPECT signs looks most promising especially when LN metastases were diagnosed in patients with enlarged solid LN or normal sized LN with Gr III-IV 99Tc-MIBI uptake. In these cases, SEN was equal to 75 %, SP-90 %, AC-85 %, only one of 5 patients with false negative results had metastases in more than 2 LN. CONCLUSIONS: By combination of SPECT and CT data we can more accurately diagnose axillary LN invasion by breast cancer.


Assuntos
Neoplasias da Mama/diagnóstico por imagem , Neoplasias da Mama/patologia , Linfonodos/diagnóstico por imagem , Tecnécio Tc 99m Sestamibi , Tomografia Computadorizada de Emissão de Fóton Único , Adulto , Idoso , Axila , Feminino , Humanos , Linfonodos/patologia , Pessoa de Meia-Idade , Invasividade Neoplásica , Estadiamento de Neoplasias , Sensibilidade e Especificidade , Tomografia Computadorizada por Raios X
20.
J Neurosci ; 34(28): 9235-48, 2014 Jul 09.
Artigo em Inglês | MEDLINE | ID: mdl-25009257

RESUMO

Precise regulation of axon branching is crucial for neuronal circuit formation, yet the mechanisms that control branch formation are not well understood. Moreover, the highly complex morphology of neurons makes them critically dependent on protein/membrane trafficking and transport systems, although the functions for membrane trafficking in neuronal morphogenesis are largely undefined. Here we identify a kinesin adaptor, Calsyntenin-1 (Clstn-1), as an essential regulator of axon branching and neuronal compartmentalization in vivo. We use morpholino knockdown and a Clstn-1 mutant to show that Clstn-1 is required for formation of peripheral but not central sensory axons, and for peripheral axon branching in zebrafish. We used live imaging of endosomal trafficking in vivo to show that Clstn-1 regulates transport of Rab5-containing endosomes from the cell body to specific locations of developing axons. Our results suggest a model in which Clstn-1 patterns separate axonal compartments and define their ability to branch by directing trafficking of specific endosomes.


Assuntos
Axônios/fisiologia , Axônios/ultraestrutura , Proteínas de Ligação ao Cálcio/metabolismo , Endossomos/fisiologia , Plasticidade Neuronal/fisiologia , Células Receptoras Sensoriais/fisiologia , Células Receptoras Sensoriais/ultraestrutura , Envelhecimento/patologia , Envelhecimento/fisiologia , Animais , Animais Geneticamente Modificados , Proteínas de Ligação ao Cálcio/genética , Células Cultivadas , Endossomos/ultraestrutura , Modelos Animais , Neurogênese/fisiologia , Transporte Proteico/fisiologia , Peixe-Zebra
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...