RESUMO
The effects of environmental radiofrequency electromagnetic fields (RF-EMF) on embryonic neural stem cells have not been determined, particularly at the proteomic level. This study aims to elucidate the effects of environmental levels of RF-EMF radiation on embryonic neural stem cells. Neuroectodermal stem cells (NE-4C cells) were randomly divided into a sham group and an RF group, which were sham-exposed and continuously exposed to a 1950 MHz RF-EMF at 2 W/kg for 48 h. After exposure, cell proliferation was determined by a Cell Counting Kit-8 (CCK8) assay, the cell cycle distribution and apoptosis were measured by flow cytometry, protein abundance was detected by liquid chromatography-tandem mass spectrometry (LC-MS/MS), and mRNA expression was evaluated by quantitative reverse transcription polymerase chain reaction (qRT-PCR). We did not detect differences in cell proliferation, cell cycle distribution, and apoptosis between the two groups. However, we detected differences in the abundance of 23 proteins between the two groups, and some of these differences were consistent with alterations in transcript levels determined by qRT-PCR (P < 0.05). A bioinformatics analysis indicated that the differentially regulated proteins were mainly enriched in 'localization' in the cellular process category; however, no significant pathway alterations in NE-4C cells were detected. We conclude that under the experimental conditions, low-level RF-EMF exposure was not neurotoxic but could induce minor changes in the abundance of some proteins involved in neurodevelopment or brain function.
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Campos Eletromagnéticos , Células-Tronco Neurais , Campos Eletromagnéticos/efeitos adversos , Cromatografia Líquida , Proteômica , Espectrometria de Massas em Tandem , Ondas de Rádio/efeitos adversosRESUMO
OBJECTIVE: 5.8 GHz spectrum is gaining more attention in wireless technology. To explore the potential hazards, we investigated the effect of exposure to 5.8 GHz microwave on learning and memory ability of rats. Methods: Morris Water maze (MWM), Novel object recognition (NOR) and Fear conditioning test (FCT) were used to evaluate the ability of spatial and non-spatial memory of rats. The hippocampal morphology, the level of brain injury factors in serum and the mitochondrial membrane potential of hippocampal neurons was examined to evaluate the damage of hippocampal neurons. The density of dendritic spines, the ultrastructure of synapses and the level of PSD95, Synaptophysin, p-CREB and CREB were detected to evaluate the hippocampal synaptic plasticity. RESULTS: Compared with Sham group, there was no significant difference in the performance of ethology (in MWM, NOR, FCT) in Microwave 2 h group or Microwave 4 h group. The hippocampal morphology, the serum level of brain injury factors and the content of mitochondrial JC-1 monomer in Microwave 2 h group or Microwave 4 h group did not change obviously, compared with Sham group. The density of dendritic spines, the ultrastructure of synapse and the level of PSD95, Synaptophysin, p-CREB and CREB in hippocampus in Microwave 2 h group or Microwave 4 h group did not significantly change, compared with Sham group. CONCLUSION: Under this experimental condition, exposure to 5.8 GHz microwave could not affect the hippocampal synaptic plasticity of rats.
Assuntos
Lesões Encefálicas , Hipocampo , Animais , Ratos , Hipocampo/metabolismo , Aprendizagem em Labirinto , Plasticidade Neuronal , Sinaptofisina/metabolismo , Sinaptofisina/farmacologiaRESUMO
Malignant tumor treatment remains a big challenge till now, and expanding literature indicated that pulsed electromagnetic fields (PEMF) is promising in tumor treatment with the advantage of safety and being economical, but it is still controversial on whether PEMF could affect the tumor cell viability. Therefore, we conducted the meta-analysis to evaluate effects of PEMF on tumor cell viability. The PubMed, EMBASE, Web of Science, and Cochrane Library databases were searched for studies published up to February 2021. Studies on the direct effects of PEMF on tumor cell viability, determined using colorimetric analysis, were included. Two authors extracted the data and completed the quality assessment. A meta-analysis was performed to calculate the absorbance values and 95% confidence intervals (CIs) using random-effects models. Seven studies, including 32 randomized controlled experiments, were analyzed. Compared with the control group, tumor cell viability in the PEMF exposure group was obviously lower (SMD, -0.67; 95% CI: -1.12 to -0.22). The subgroup meta-analysis results showed that PEMF significantly reduced epithelial cancer cell viability (SMD, -0.58; 95% CI: -0.92 to -0.23) but had no influence on stromal tumor cell viability (SMD, -0.93; 95% CI: -0.21 to 0.15). Our study demonstrated that PEMF could inhibit tumor cell proliferation to some extent, but the risk of bias and high heterogeneity (I2 > 75%) weakened the strength of the conclusions drawn from the analysis.
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Campos Eletromagnéticos , Neoplasias , Proliferação de Células , Sobrevivência Celular , HumanosRESUMO
BACKGROUND: To investigate the functions of the hyperpolarization-activated cation currents in medium-size dorsal root ganglion cells in a rat model of overactive bladder syndrome. METHODS: Rats with OAB were screened using a urodynamic testing device. The whole-cell patch clamp technique was used to investigate changes in excitability and hyperpolarization-activated cation current (Ih) of medium-size cells in the L6 dorsal root ganglia (DRG) of the OAB rats. Intrathecal injection of the specific Ih inhibitor ZD7288 was used to investigate changes of voiding function and Ih of medium-size cells in the L6 DRG. RESULTS: The urinary bladder weight of the OAB rats was significantly increased (p < 0.01); However, 7 days after intrathecally administration of ZD7288 (2 µM), the weight of rat bladder was significantly reduced (p < 0.01). The excitability of the medium-size cells in the L6 DRG of the OAB rats was significantly increased, and the number of action potentials elicited by a 500 pA stimulus was also markedly increased. Furthermore, ZD7288 significantly reduced the excitability of the medium-size DRG cells. The medium-size cells in the DRG of the OAB rats had a significantly increased Ih current density, which was blocked by ZD7288. CONCLUSIONS: The Ih current density significantly increased in medium-size cells of the L6 DRG in the OAB model. A decrease of the Ih current was able to significantly improve the voiding function of the OAB rats, in addition to lowering their urinary bladder weight. Our finding suggested that the observed increase of Ih current in the medium-size DRG neurons might play an important role in the pathological processes of OAB.
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Potenciais de Ação , Gânglios Espinais/citologia , Bexiga Urinária Hiperativa/fisiopatologia , Animais , Cátions , Feminino , Ratos , SíndromeRESUMO
BACKGROUND: Transcranial direct current stimulation (tDCS) is a noninvasive neural control technology that has become a research hotspot. To facilitate further research of tDCS, the biosafety of 500 µA cathodal tDCS, a controversial parameter in rats was evaluated. RESULTS: 24 animals were randomly divided into two groups: a cathodal tDCS group (tDCS, n = 12) and control group (control, n = 12). Animals in the tDCS group received 5 consecutive days of cathodal tDCS (500 µA, 15 min, once per day) followed by a tDCS-free interval of 2 days and 5 additional days of stimulation, totally two treatments of tDCS for a total of 10 days. Computational 3D rat model was adopted to calculate the current density distributions in brain during tDCS treatment. Essential brain functions including motor function and learning and memory ability were evaluated. Additionally, to estimate the neurotoxicity of tDCS, the brain morphology, neurotransmitter levels and cerebral temperature were investigated. Our results showed that the current density inside the brain was less than 20 A/m2 during tDCS treatment in computational model. tDCS did not affect motor functions and learning and memory ability after tDCS treatment. In addition, no significant differences were found for the tDCS group in hematology, serum biochemical markers or the morphology of major organs. Moreover, tDCS treatment had no effect on the brain morphology, neural structures, neurotransmitter levels or cerebral temperature. CONCLUSION: 500 µA cathodal tDCS as performed in the present study was safe for rodents.
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Biomarcadores/sangue , Aprendizagem/fisiologia , Memória/fisiologia , Atividade Motora/fisiologia , Córtex Motor/fisiologia , Segurança , Estimulação Transcraniana por Corrente Contínua/efeitos adversos , Animais , Simulação por Computador , Hipocampo/metabolismo , Testes de Função Renal , Testes de Função Hepática , Masculino , Córtex Motor/metabolismo , Córtex Motor/patologia , Neurotransmissores/metabolismo , Ratos , Teste de Desempenho do Rota-Rod , TemperaturaRESUMO
OBJECTIVE: The study aimed to report long-term outcomes of facial nerve schwannomas (FNS) with favorable facial nerve function by observation, and to discuss about the relationship between initial tumor size and tumor growth. METHODS: 21 facial nerve schwannoma cases with favorable facial nerve function were managed by observation. They were divided into larger size group (size ≥10mm) and smaller size group (size <10mm) according to initial tumor size. RESULTS: They were followed up for 6.4±1.7years. 18 of 21 cases (85.7%) maintained House-Brackmann Grade III or better. Growth rate of the tumors in larger size group was 72.7%, much higher than 10% in smaller size group (p<0.05). CONCLUSIONS: Observation was feasible for most FNS with favorable facial nerve function, and growth rate of the tumors was associated with tumor size.
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Neoplasias dos Nervos Cranianos/patologia , Neoplasias dos Nervos Cranianos/terapia , Nervo Facial/patologia , Neurilemoma/patologia , Neurilemoma/terapia , Conduta Expectante/métodos , Adulto , Fatores Etários , Idoso , China , Estudos de Coortes , Neoplasias dos Nervos Cranianos/cirurgia , Progressão da Doença , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Invasividade Neoplásica/patologia , Estadiamento de Neoplasias , Neurilemoma/cirurgia , Prognóstico , Estudos Retrospectivos , Medição de Risco , Fatores Sexuais , Fatores de Tempo , Carga Tumoral , Adulto JovemRESUMO
Mesencephalic trigeminal nucleus (Mes V) neurons represent an uncommon class of primary sensory neurons. Besides receiving somatosensory information, Mes V neurons are also involved in regulating multisensory information. The present review first describes the passive features as well as three important currents, followed by a distinct excitability classification and a description of the excitability transition of Mes V neurons. Furthermore, their resonance property, the existence of membrane oscillation and electrical coupling which may promote strong synchronization, as well as their function in controlling stretch reflex activity, are discussed.
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Fenômenos Eletrofisiológicos/fisiologia , Mesencéfalo/fisiologia , Células Receptoras Sensoriais/fisiologia , Núcleos do Trigêmeo/fisiologia , Animais , Humanos , Mesencéfalo/citologia , Células Receptoras Sensoriais/citologia , Núcleos do Trigêmeo/citologiaRESUMO
BACKGROUND: Sympathetic ganglion block (SGB) technique is becoming increasingly prevalent in the treatment of complex regional pain syndromes (CRPS). Given the varied reported effectiveness of these techniques and the heterogeneity of treatment regimens, there is an urgent need for consistent and high-quality evidence on the efficacy and safety of such procedures. OBJECTIVES: This study aimed to compare the efficacy of SGB therapy for CRPS-related pain. STUDY DESIGN: A meta-analysis of randomized controlled trials (RCTs). METHODS: PubMed, EMBASE, Web of Science, CINAHL, US National Institutes of Health Clinical Trials Registry, Google Scholar, and Cochrane Library Databases were systematically searched between January 1967 and April 2023. A meta-analysis of the included RCTs on SGB was conducted to evaluate the effectiveness and risk of bias (ROBs) of SGB. RESULTS: After screening 8523 records, 12 RCTs were included in this meta-analysis. Compared with controls, the visual analog pain score decreased by a weighted mean difference (WMD) of -6.24 mm (95% CI, -11.45, -1.03; P = 0.019) in the random-effects model, and the numerical scale score was reduced by a WMD of -1.17 mm (95% CI, -2.42, 0.08; P = 0.067) in the fixed-effects model, indicating a pain relief. The methodological quality of the included RCTs was high, with an average PEDro score of 7.0 (range: 5-9). LIMITATIONS: The number of included trials was limited. CONCLUSIONS: SGB therapy can reduce pain intensity in patients with CRPS with few adverse events. However, owing to the relatively high heterogeneity of the included RCTs, a larger sample of high-quality RCTs is needed to further confirm this conclusion.
Assuntos
Bloqueio Nervoso Autônomo , Síndromes da Dor Regional Complexa , Gânglio Estrelado , Humanos , Síndromes da Dor Regional Complexa/terapia , Bloqueio Nervoso Autônomo/métodos , Ensaios Clínicos Controlados Aleatórios como AssuntoRESUMO
Painful diabetic neuropathy is a common complication of diabetes mellitus and can affect many aspects of life and severely limit patients' daily functions. Signals of painful diabetic neuropathy are believed to originate in the peripheral nervous system. However, its peripheral mechanism of hyperalgesia has remained elusive. Numerous studies have accumulated that polymodal nociceptive C-fibres play a crucial role in the generation and conduction of pain signals and sensitization of which following injury or inflammation leads to marked hyperalgesia. Traditionally, the number of nociceptive primary afferent firings is believed to be determined at the free nerve endings, while the extended main axon of unmyelinated C-fibres only involves the reliable and faithful propagation of firing series to the central terminals. We challenged this classic view by showing that conduction of action potential can fail to occur in response to repetitive activity when they travel down the main axon of polymodal nociceptive C-fibres. Quantitative analysis of conduction failure revealed that the degree of conduction failure displays a frequency-dependent manner. Local administration of low threshold, rapidly activating potassium current blocker, α-dendrotoxin (0.5 nM) and persistent sodium current blocker, low doses of tetrodotoxin (<100 nM) on the main axon of C-fibres can reciprocally regulate the degree of conduction failure, confirming that conduction failure did occur along the main axon of polymodal nociceptive C-fibres. Following streptozotocin-induced diabetes, a subset of polymodal nociceptive C-fibres exhibited high-firing-frequency to suprathreshold mechanical stimulation, which account for about one-third of the whole population of polymodal nociceptive C-fibres tested. These high-firing-frequency polymodal nociceptive C-fibres in rats with diabetes displayed a marked reduction of conduction failure. Delivery of low concentrations of tetrodotoxin and Nav1.8 selective blocker, A-803467 on the main axon of C-fibres was found to markedly enhance the conduction failure in a dose-dependent manner in diabetic rats. Upregulated expression of sodium channel subunits Nav1.7 and Nav1.8 in both small dorsal root ganglion neurons and peripheral C-fibres as well as enhanced transient and persistent sodium current and increased excitability in small dorsal root ganglion neurons from diabetic rats might underlie the reduced conduction failure in the diabetic high-firing-frequency polymodal nociceptive C-fibres. This study shed new light on the functional capability in the pain signals processing for the main axon of polymodal nociceptive C-fibres and revealed a novel mechanism underlying diabetic hyperalgesia.
Assuntos
Axônios/fisiologia , Neuropatias Diabéticas/fisiopatologia , Hiperalgesia/fisiopatologia , Condução Nervosa/fisiologia , Nociceptores/fisiologia , Potenciais de Ação/fisiologia , Animais , Masculino , Fibras Nervosas/fisiologia , Ratos , Ratos Sprague-DawleyRESUMO
This study was designed to explore the functional circuitry of the adult zebrafish cerebellum, focusing on its Purkinje cells and using whole-cell patch recordings and single cell labeling in slice preparations. Following physiological characterizations, the recorded single cells were labeled for morphological identification. It was found that the zebrafish Purkinje cells are surprisingly diverse. Based on their physiology and morphology, they can be classified into at least three subtypes: Type I, a narrow spike cell, which fires only narrow Na+ spikes (<3 ms in duration), and has a single primary dendrite with an arbor restricted to the distal molecular layer; Type II, a broad spike cell, which fires broad Ca2+ spikes (5-7 ms in duration) and has a primary dendrite with limited branching in the inner molecular layer and then further radiates throughout the molecular layer; and Type III, a very broad spike cell, which fires very broad Ca2+ spikes (≥10 ms in duration) and has a dense proximal dendritic arbor that is either restricted to the inner molecular layer (Type IIIa), or radiates throughout the entire molecular layer (Type IIIb). The graded paired-pulse facilitation of these Purkinje cells' responses to parallel fiber activations and the all-or-none, paired-pulse depression of climbing fiber activation are largely similar to those reported for mammals. The labeled axon terminals of these Purkinje cells end locally, as reported for larval zebrafish. The present study provides evidence that the corresponding functional circuitry and information processing differ from what has been well-established in the mammalian cerebellum.
Assuntos
Células de Purkinje , Peixe-Zebra , Animais , Células de Purkinje/fisiologia , Peixe-Zebra/fisiologia , Potenciais de Ação/fisiologia , Cerebelo , Axônios/fisiologia , MamíferosRESUMO
OBJECTIVES: Recent evidence indicates that hippocampus is important for conditioned fear memory (CFM). Though few studies consider the roles of various cell types' contribution to such a process, as well as the accompanying transcriptome changes during this process. The purpose of this study was to explore the transcriptional regulatory genes and the targeted cells that are altered by CFM reconsolidation. METHODS: A fear conditioning experiment was established on adult male C57 mice, after day 3 tone-cued CFM reconsolidation test, hippocampus cells were dissociated. Using single cell RNA sequencing (scRNA-seq) technique, alterations of transcriptional genes expression were detected and cell cluster analysis were performed and compared with those in sham group. RESULTS: Seven non-neuronal and eight neuronal cell clusters (including four known neurons and four newly identified neuronal subtypes) has been explored. Among them, CA subtype 1 has characteristic gene markers of Ttr and Ptgds, which is speculated to be the outcome of acute stress and promotes the production of CFM. The results of KEGG pathway enrichment indicate the differences in the expression of certain molecular protein functional subunits in long-term potentiation (LTP) pathway between two types of neurons (DG and CA1) and astrocytes, thus providing a new transcriptional perspective for the role of hippocampus in the CFM reconsolidation. More importantly, the correlation between the reconsolidation of CFM and neurodegenerative diseases-linked genes is substantiated by the results from cell-cell interactions and KEGG pathway enrichment. Further analysis shows that the reconsolidation of CFM inhibits the risk-factor genes App and ApoE in Alzheimer's Disease (AD) and activates the protective gene Lrp1. CONCLUSIONS: This study reports the transcriptional genes expression changes of hippocampal cells driven by CFM, which confirm the involvement of LTP pathway and suggest the possibility of CFM-like behavior in preventing AD. However, the current research is limited to normal C57 mice, and further studies on AD model mice are needed to prove this preliminary conclusion.
Assuntos
Hipocampo , Transtornos Fóbicos , Camundongos , Masculino , Animais , Hipocampo/metabolismo , Neurônios/fisiologia , Sinais (Psicologia) , Medo/fisiologiaRESUMO
Light exposure can profoundly affect neurological functions and behaviors. Here, we show that short-term exposure to moderate (400 lux) white light during Y-maze test promoted spatial memory retrieval and induced only mild anxiety in mice. This beneficial effect involves the activation of a circuit including neurons in the central amygdala (CeA), locus coeruleus (LC), and dentate gyrus (DG). Specifically, moderate light activated corticotropin-releasing hormone (CRH) positive (+) CeA neurons and induced the release of corticotropin-releasing factor (CRF) from their axon terminals ending in the LC. CRF then activated tyrosine hydroxylase-expressing LC neurons, which send projections to DG and release norepinephrine (NE). NE activated ß-adrenergic receptors on CaMKIIα-expressing DG neurons, ultimately promoting spatial memory retrieval. Our study thus demonstrated a specific light scheme that can promote spatial memory without excessive stress, and unraveled the underlying CeA-LC-DG circuit and associated neurochemical mechanisms.
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Tonsila do Cerebelo , Luz , Memória Espacial , Tonsila do Cerebelo/citologia , Tonsila do Cerebelo/metabolismo , Animais , Camundongos , Ansiedade , Giro Denteado/citologia , Giro Denteado/metabolismo , Neurônios , Locus Cerúleo/citologia , Locus Cerúleo/metabolismo , Hormônio Liberador da Corticotropina/metabolismo , Norepinefrina/metabolismo , Vias Neurais , Aprendizagem em Labirinto , Camundongos Endogâmicos C57BLRESUMO
The adult cortex has long been regarded as non-neurogenic. Whether injury can induce neurogenesis in the adult cortex is still controversial. Here, we report that focal ischemia stimulates a transient wave of local neurogenesis. Using 5'-bromo-2'-deoxyuridine labeling, we demonstrated a rapid generation of doublecortin-positive neuroblasts that died quickly in mouse cerebral cortex following ischemia. Nestin-CreER-based cell ablation and fate mapping showed a small contribution of neuroblasts by subventricular zone neural stem cells. Using a mini-photothrombotic ischemia mouse model and retrovirus expressing green fluorescent protein labeling, we observed maturation of locally generated new neurons. Furthermore, fate tracing analyses using PDGFRα-, GFAP-, and Sox2-CreER mice showed a transient wave of neuroblast generation in mild ischemic cortex and identified that Sox2-positive astrocytes were the major neurogenic cells in adult cortex. In addition, a similar upregulation of Sox2 and appearance of neuroblasts were observed in the focal ischemic cortex of Macaca mulatta. Our findings demonstrated a transient neurogenic response of Sox2-positive astrocytes in ischemic cortex, which suggests the possibility of inducing neuronal regeneration by amplifying this intrinsic response in the future.
RESUMO
Reactive astrogliosis usually bears some properties of neural progenitors. How injury triggers astrocyte dedifferentiation remains largely unclear. Here, we report that ischemia induces rapid up-regulation of Wnt2 protein in apoptotic neurons and activation of canonical Wnt signaling in reactive astrocytes in mice, primates and human. Local delivery of Wnt2 shRNA abolished the dedifferentiation of astrocytes while over-expressing Wnt2 promoted progenitor marker expression and neurogenesis. Both the activation of Wnt signaling and dedifferentiation of astrocytes was compromised in ischemic caspase-3-/- cortex. Over-expressing stabilized ß-catenin not only facilitated neurogenesis but also promoted functional recovery in ischemic caspase-3-/- mice. Further analysis showed that apoptotic neurons up-regulated Wnt2 protein via internal ribosome entry site (IRES)-mediated translation. Knocking down death associated protein 5 (DAP5), a key protein in IRES-mediated protein translation, significantly diminished Wnt activation and astrocyte dedifferentiation. Our data demonstrated an apoptosis-initiated Wnt-activating mechanism which triggers astrocytic dedifferentiation and facilitates neuronal regeneration.
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Noise can play a constructive role in the detection of weak signals in various kinds of peripheral receptors and neurons. What the mechanism underlying the effect of noise is remains unclear. Here, the perforated patch-clamp technique was used on isolated cells from chronic compression of the dorsal root ganglion (DRG) model. Our data provided new insight indicating that, under conditions without external signals, noise can enhance subthreshold oscillations, which was observed in a certain type of neurons with high-frequency (20-100 Hz) intrinsic resonance from injured DRG neurons. The occurrence of subthreshold oscillation considerably decreased the threshold potential for generating repetitive firing. The above effects of noise can be abolished by blocking the persistent sodium current (I(Na, P)). Utilizing a mathematical neuron model we further simulated the effect of noise on subthreshold oscillation and firing, and also found that noise can enhance the electrical activity through autonomous stochastic resonance. Accordingly, we propose a new concept of the effects of noise on neural intrinsic activity, which suggests that noise may be an important factor for modulating the excitability of neurons and generation of chronic pain signals.
Assuntos
Relógios Biológicos/fisiologia , Gânglios Espinais/patologia , Ruído , Radiculopatia/patologia , Células Receptoras Sensoriais/fisiologia , Potenciais de Ação/fisiologia , Animais , Células Cultivadas , Modelos Animais de Doenças , Estimulação Elétrica/métodos , Matemática , Modelos Neurológicos , Técnicas de Patch-Clamp , Ratos , Ratos Sprague-Dawley , Células Receptoras Sensoriais/classificação , Células Receptoras Sensoriais/efeitos dos fármacos , Bloqueadores dos Canais de Sódio/farmacologia , Tetrodotoxina/farmacologiaRESUMO
BACKGROUND: Sympathetically maintained pain (SMP) involves an increased excitability of dorsal root ganglion (DRG) neurons to sympathetic nerve stimulation and circulating norepinephrine. The current treatment of SMP has limited efficacy, and hence more mechanistic insights into this intractable pain condition are urgently needed. METHODS: A caudal trunk transection (CTT) model of neuropathic pain was established in mice.Immunofluorescence staining, small interfering RNA, pharmacological and electrophysiological studies were conducted to test the hypothesis that norepinephrine increases the excitability of small-diameter DRG neurons from CTT mice through the activation of cyclic guanosine monophosphate-protein kinase G (cGMP-PKG) signaling pathway. RESULTS: Behavior study showed that CTT mice developed mechanical and heat hypersensitivities, which were attenuated by intraperitoneal injection of guanethidine. CTT mice also showed an abnormal sprouting of tyrosine hydroxylase-positive nerve fibers in DRG, and an increased excitability of small-diameter DRG neurons to norepinephrine, suggesting that CTT is a useful model to study SMP. Importantly, inhibiting cGMP-PKG pathway with small interfering RNA and KT5823 attenuated the increased sympathetic sensitivity in CTT mice. In contrast, cGMP activators (Sp-cGMP, 8-Br-cGMP) further increased sympathetic sensitivity. Furthermore, phosphorylation of ATP-sensitive potassium channel, which is a downstream target of PKG, may contribute to the adrenergic modulation of DRG neuron excitability. CONCLUSIONS: Our findings suggest an important role of cGMP-PKG signaling pathway in the increased excitability of small-diameter DRG neurons to norepinephrine after CTT, which involves an inhibition of the ATP-sensitive potassium currents through PKG-induced phosphorylation. Accordingly, drugs targeting this pathway may help to treat SMP.
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Proteínas Quinases Dependentes de GMP Cíclico , Neuralgia , Animais , Proteínas Quinases Dependentes de GMP Cíclico/metabolismo , Gânglios Espinais/metabolismo , Canais KATP/metabolismo , Camundongos , Transdução de SinaisRESUMO
Background. Ischemic stroke carries a high mortality rate and is a leading cause of severe neurological disability. However, the efficacy of current therapeutic options remains limited. Objective. We aimed to investigate the treatment efficacy of transcranial direct current stimulation (tDCS) in motor function rehabilitation after ischemic stroke and explore the underlying mechanisms. Methods. Male Sprague-Dawley rats with epicranial electrodes were used to establish pathogenetic model through temporary right middle cerebral artery occlusion (MCAO). Subsequently, animals were randomly divided into 4 groups: MCAO + tDCS/sham tDCS, Control + tDCS/sham tDCS. Animals in the groups with tDCS underwent 10 days of cathodal tDCS totally (500 µA, 15 minutes, once a day). During and after tDCS treatment, the motor functions of the animals, ischemic damage area, proliferation and differentiation of neural stem cells (NSCs), and distribution, and protein expression of Notch1 signaling molecules were detected. Results. The rehabilitation of MCAO-induced motor function deficits was dramatically accelerated by tDCS treatment. NSC proliferation in the subventricular zone (SVZ) was significantly increased after MCAO surgery, and tDCS treatment promoted this process. Additionally, NSCs probably migrated from the SVZ to the ischemic striatum and then differentiated into neurons and oligodendrocytes after MCAO surgery, both of which processes were accelerated by tDCS treatment. Finally, tDCS treatment inhibited the activation of Notch1 signaling in NSCs in the ischemic striatum, which may be involved in NSC differentiation in the MCAO model. Conclusion. Our results suggest that tDCS may exert therapeutic efficacy after ischemic stroke in a regenerative medical perspective.
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AVC Isquêmico/fisiopatologia , AVC Isquêmico/reabilitação , Atividade Motora/fisiologia , Neurogênese/fisiologia , Receptor Notch1/metabolismo , Recuperação de Função Fisiológica/fisiologia , Reabilitação do Acidente Vascular Cerebral , Estimulação Transcraniana por Corrente Contínua , Animais , Comportamento Animal/fisiologia , Modelos Animais de Doenças , Infarto da Artéria Cerebral Média/complicações , AVC Isquêmico/etiologia , Masculino , Células-Tronco Neurais/fisiologia , Ratos , Ratos Sprague-Dawley , Transdução de Sinais/fisiologiaRESUMO
Previous studies have shown that CCL2 (C-C motif chemokine ligand 2) induces chronic pain, but the exact mechanisms are still unknown. Here, we established models to explore the potential mechanisms. Behavioral experiments revealed that an antagonist of extracellular signal-regulated kinase (ERK) inhibited not only CCL2-induced inflammatory pain, but also pain responses induced by complete Freund's adjuvant. We posed the question of the intracellular signaling cascade involved. Subsequent experiments showed that CCL2 up-regulated the expression of phosphorylated ERK (pERK) and N-methyl D-aspartate receptor [NMDAR] subtype 2B (GluN2B); meanwhile, antagonists of CCR2 and ERK effectively reversed these phenomena. Whole-cell patch-clamp recordings revealed that CCL2 enhanced the NMDAR-induced currents via activating the pERK pathway, which was blocked by antagonists of GluN2B and ERK. In summary, we demonstrate that CCL2 directly interacts with CCR2 to enhance NMDAR-induced currents, eventually leading to inflammatory pain mainly through the CCL2-CCR2-pERK-GluN2B pathway.
Assuntos
Quimiocina CCL2/metabolismo , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , N-Metilaspartato , Dor , Receptores de N-Metil-D-Aspartato/metabolismo , Substância Gelatinosa/fisiologia , Animais , Quimiocina CCL2/antagonistas & inibidores , MAP Quinases Reguladas por Sinal Extracelular/antagonistas & inibidores , Masculino , Camundongos , Camundongos Endogâmicos C57BL , N-Metilaspartato/metabolismo , Neurônios , Transdução de SinaisRESUMO
Recent experimental and theoretical data indicate that the functional capabilities of axons with specialized structures are much more diverse than traditionally thought. However, few observations were concerned with the main axons without arborization. In the present study, electrical stimulation of the saphenous nerve at different frequencies (2, 5, 10, 20 Hz) was used to test the role of activity-dependent effects on the pattern of action potentials that propagate along individual unmyelinated fibers (C fibers) within the trunk of the saphenous nerve in rabbits. Three basic types of C fiber responses to repetitive stimulation were observed: type-1 fibers showed an entrained response without conduction failure; type-2 fibers discharged with intermittent conduction failures; while only sporadic conduction failures happened in type 3. The failure modality in type-2 and type-3 fibers is closely related to the conductive distance as well as the frequency and duration of stimuli which lead to a critical level of conduction velocity slowing. A novel fluctuation in interspike intervals was always observed immediately before the occurrence of the failures, implying that the fluctuation of conduction velocity is correlated with imminent failures. Both the 4-aminopyridine-sensitive potassium current and hyperpolarization-activated cation current were recognized to be involved in the regulation of conduction failure patterns. The results confirmed, at least in part, the existence of conduction failures in the main axon of C fibers, suggesting that axonal operations may also be determinants for adaptation phenomenon and information processing in peripheral nervous system.
Assuntos
Potenciais de Ação/fisiologia , Axônios/fisiologia , Nervo Femoral/fisiologia , Fibras Nervosas Amielínicas/fisiologia , Condução Nervosa/fisiologia , Sistema Nervoso Periférico/fisiologia , 4-Aminopiridina/farmacologia , Potenciais de Ação/efeitos dos fármacos , Animais , Axônios/efeitos dos fármacos , Axônios/ultraestrutura , Membrana Celular/efeitos dos fármacos , Membrana Celular/fisiologia , Relação Dose-Resposta a Droga , Estimulação Elétrica , Feminino , Nervo Femoral/citologia , Ativação do Canal Iônico/efeitos dos fármacos , Ativação do Canal Iônico/fisiologia , Masculino , Fibras Nervosas Amielínicas/efeitos dos fármacos , Fibras Nervosas Amielínicas/ultraestrutura , Condução Nervosa/efeitos dos fármacos , Sistema Nervoso Periférico/citologia , Bloqueadores dos Canais de Potássio/farmacologia , Canais de Potássio/efeitos dos fármacos , Canais de Potássio/fisiologia , Coelhos , Tempo de Reação/efeitos dos fármacos , Tempo de Reação/fisiologia , Fatores de TempoRESUMO
Single-fiber recording has been a classical and effective electrophysiological technique over the last few decades because of its specific application for nerve fibers in the central and peripheral nervous systems. This method is particularly applicable to dorsal root ganglia (DRG), which are primary sensory neurons that exhibit a pseudo-unipolar structure of nervous processes. The patterns and features of the action potentials passed along axons are recordable in these neurons. The present study uses in vivo single-fiber recordings to observe the conduction failure of sciatic nerves in complete Freund's adjuvant (CFA)-treated rats. As the underlying mechanism cannot be studied using in vivo single-fiber recordings, patch-clamp-recordings of DRG neurons are performed on preparations of intact DRG with the attached sciatic nerve. These recordings reveal a positive correlation between conduction failure and the rising slope of the after-hyperpolarization potential (AHP) of DRG neurons in CFA-treated animals. The protocol for in vivo single fiber-recordings allows the classification of nerve fibers via the measurement of conduction velocity and monitoring of abnormal conditions in nerve fibers in certain diseases. Intact DRG with attached peripheral nerve allows observation of the activity of DRG neurons in most physiological conditions. Conclusively, single-fiber recording combined with electrophysiological recording of intact DRGs is an effective method to examine the role of conduction failure during the analgesic process.