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The dorsal raphe nucleus (DRN) receives dopaminergic inputs from the ventral tegmental area (VTA). Also, the DRN contains a small population of cells that express dopamine (DRNDA neurons). However, the physiological role of dopamine (DA) in the DRN and its interaction with serotonergic (5-HT) neurons is poorly understood. Several works have reported moderate levels of D1, D2, and D3 DA receptors in the DRN. Furthermore, it was found that the activation of D2 receptors increased the firing of putative 5-HT neurons. Other studies have reported that D1 and D2 dopamine receptors can interact with glutamate NMDA receptors, modulating the excitability of different cell types. In the present work, we used immunocytochemical techniques to determine the kind of DA receptors in the DRN. Additionally, we performed electrophysiological experiments in brainstem slices to study the effect of DA agonists on NMDA-elicited currents recorded from identified 5-HT DRN neurons. We found that D2 and D3 but not D1 receptors are present in this nucleus. Also, we demonstrated that the activation of D2-like receptors increases NMDA-elicited currents in 5-HT neurons through a mechanism involving phospholipase C (PLC) and protein kinase C (PKC) enzymes. Possible physiological implications related to the sleep-wake cycle are discussed.
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Núcleo Dorsal da Rafe , Receptores de Dopamina D2 , Receptores de N-Metil-D-Aspartato , Neurônios Serotoninérgicos , Animais , Núcleo Dorsal da Rafe/metabolismo , Núcleo Dorsal da Rafe/efeitos dos fármacos , Receptores de Dopamina D2/metabolismo , Neurônios Serotoninérgicos/metabolismo , Neurônios Serotoninérgicos/efeitos dos fármacos , Neurônios Serotoninérgicos/fisiologia , Masculino , Receptores de N-Metil-D-Aspartato/metabolismo , Receptores de Dopamina D3/metabolismo , N-Metilaspartato/farmacologia , N-Metilaspartato/metabolismo , Receptores de Dopamina D1/metabolismo , Receptores de Dopamina D1/agonistas , Agonistas de Dopamina/farmacologia , Ratos , Fosfolipases Tipo C/metabolismo , Ratos WistarRESUMO
The dorsal motor nucleus of the vagus (DMV) contains parasympathetic motoneurons that project to the heart and lungs. These motoneurons control ventricular excitability/contractility and airways secretions/blood flow, respectively. However, their electrophysiological properties, morphology and synaptic input activity remain unknown. One important ionic current described in DMV motoneurons controlling their electrophysiological behaviour is the A-type mediated by voltage-dependent K+ (Kv) channels. Thus, we compared the electrophysiological properties, synaptic activity, morphology, A-type current density, and single cell expression of Kv subunits, that contribute to macroscopic A-type currents, between DMV motoneurons projecting to either the heart or lungs of adult male rats. Using retrograde labelling, we visualized distinct DMV motoneurons projecting to the heart or lungs in acutely prepared medullary slices. Subsequently, whole cell recordings, morphological reconstruction and single motoneuron qRT-PCR studies were performed. DMV pulmonary motoneurons were more depolarized, electrically excitable, presented higher membrane resistance, broader action potentials and received greater excitatory synaptic inputs compared to cardiac DMV motoneurons. These differences were in part due to highly branched dendritic complexity and lower magnitude of A-type K+ currents. By evaluating expression of channels that mediate A-type currents from single motoneurons, we demonstrated a lower level of Kv4.2 in pulmonary versus cardiac motoneurons, whereas Kv4.3 and Kv1.4 levels were similar. Thus, with the distinct electrical, morphological, and molecular properties of DMV cardiac and pulmonary motoneurons, we surmise that these cells offer a new vista of opportunities for genetic manipulation providing improvement of parasympathetic function in cardiorespiratory diseases such heart failure and asthma.
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Coração , Pulmão , Neurônios Motores , Nervo Vago , Animais , Neurônios Motores/fisiologia , Masculino , Coração/fisiologia , Coração/inervação , Pulmão/fisiologia , Pulmão/inervação , Nervo Vago/fisiologia , Bulbo/fisiologia , Bulbo/citologia , Bulbo/metabolismo , Potenciais de Ação/fisiologia , Ratos Sprague-Dawley , Ratos , Técnicas de Patch-ClampRESUMO
The East Pacific (EP) region, especially the central and southern EP, has been fairly less studied than other world's regions with respect to marine litter pollution. This comprehensive literature review (257 peer-reviewed publications) showed that both macrolitter (mostly plastics) and microplastics tend to accumulate on EP shorelines. Moreover, they were also reported in all the other compartments investigated: sea surface, water column, seafloor and 'others'. Mostly local, land-based sources (e.g., tourism, poor waste management) were identified across the region, especially at continental sites from low and mid latitudes. Some sea-based sources (e.g., fisheries, long-distance drifting) were also identified at high latitudes and on oceanic islands, likely enhanced by the oceanographic dynamics of the EP that affect transport of floating litter. Our results suggest that effective solutions to the problem require local and preventive strategies to significantly reduce the levels of litter along the EP coasts.
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Monitoramento Ambiental , Microplásticos , Plásticos , Poluentes Químicos da Água , Microplásticos/análise , Plásticos/análise , Poluentes Químicos da Água/análise , Oceano PacíficoRESUMO
Numerous studies suggest the involvement of adenosine-5'-triphosphate (ATP) and similar nucleotides in the pathophysiology of asthma. Androgens, such as testosterone (TES), are proposed to alleviate asthma symptoms in young men. ATP and uridine-5'-triphosphate (UTP) relax the airway smooth muscle (ASM) via purinergic P2Y2 and P2Y4 receptors and K+ channel opening. We previously demonstrated that TES increased the expression of voltage-dependent K+ (KV) channels in ASM. This study investigates how TES may potentiate ASM relaxation induced by ATP and UTP. Tracheal tissues treated with or without TES (control group) from young male guinea pigs were used. In organ baths, tracheas exposed to TES (40 nM for 48 h) showed enhanced ATP- and UTP-evoked relaxation. Tetraethylammonium, a K+ channel blocker, annulled this effect. Patch-clamp experiments in tracheal myocytes showed that TES also increased ATP- and UTP-induced K+ currents, and this effect was abolished with flutamide (an androgen receptor antagonist). KV channels were involved in this phenomenon, which was demonstrated by inhibition with 4-aminopyridine. RB2 (an antagonist of almost all P2Y receptors except for P2Y2), as well as N-ethylmaleimide and SQ 22,536 (inhibitors of G proteins and adenylyl cyclase, respectively), attenuated the enhancement of the K+ currents induced by TES. Immunofluorescence and immunohistochemistry studies revealed that TES did not modify the expression of P2Y4 receptors or COX-1 and COX-2, while we have demonstrated that this androgen augmented the expression of KV1.2 and KV1.5 channels in ASM. Thus, TES leads to the upregulation of P2Y4 signaling and KV channels in guinea pig ASM, enhancing ATP and UTP relaxation responses, which likely limits the severity of bronchospasm in young males.
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Trifosfato de Adenosina , Adenilil Ciclases , Relaxamento Muscular , Músculo Liso , Testosterona , Traqueia , Uridina Trifosfato , Animais , Uridina Trifosfato/farmacologia , Uridina Trifosfato/metabolismo , Cobaias , Relaxamento Muscular/efeitos dos fármacos , Masculino , Trifosfato de Adenosina/metabolismo , Traqueia/metabolismo , Traqueia/efeitos dos fármacos , Testosterona/farmacologia , Testosterona/metabolismo , Adenilil Ciclases/metabolismo , Músculo Liso/metabolismo , Músculo Liso/efeitos dos fármacos , Canais de Potássio de Abertura Dependente da Tensão da Membrana/metabolismo , Transdução de Sinais/efeitos dos fármacos , Receptores Purinérgicos P2/metabolismoRESUMO
OBJECTIVE: To investigate the effect of diadynamic currents administered prior to exercises on pain and disability in patients with osteoarthritis of the knee. DESIGN: A randomized-controlled trial. SETTING: Special Rehabilitation Services in Taboão da Serra. PARTICIPANTS: Patients with bilateral knee osteoarthritis. INTERVENTION: Participants were randomly allocated to Group I (diadynamic currents and exercises; n = 30, 60 knees) or Group II (exercises alone; n = 30, 60 knees) and were treated three times a week for 8 weeks. MAIN OUTCOME MEASURES: The primary outcome measures were change in knee pain evaluated by visual analog scale and disability Index Score (Lequesne). Secondary outcomes included change in mobility (Timed Up and Go test), range of motion (goniometer), muscle strength (dynamometer), a composite score for pain and disability (Western Ontario and McMaster Universities Osteoarthritis questionnaire), and a drug diary to measure consumption of rescue pain medication (paracetamol). All measurements were collected at baseline, 8 weeks, and 6 months from baseline (follow-up). RESULTS: There were 60 participants with a mean (SD) age of 63.40 (8.20) years. Between-group differences in the follow-up (8 weeks and 6 months) were observed for pain at rest, pain during activities of daily living and disability. There was improvement in Group I that was maintained for the three variables 6 months after treatment. Mean difference for pain at rest was -3.08 points (95% confidence interval -4.13; -2.02), p < 0.01 with an effect size of 1.4; mean difference for pain during activities of daily living was -2.40 points (95% confidence interval -3.34; -1.45), p < 0.01 with an effect size of 1.24; and mean difference for disability was -4.08 points (95% confidence interval -5.89; -2.26), p < 0.01 with an effect size of 1.04. CONCLUSION: Patients with symptomatic knee osteoarthritis receiving 8 weeks of treatment with diadynamic currents as an adjunct to a program of exercises had significantly greater improvements in pain and disability than those receiving exercises alone. Beneficial effects were sustained for 6 months.
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Terapia por Exercício , Osteoartrite do Joelho , Medição da Dor , Humanos , Osteoartrite do Joelho/reabilitação , Osteoartrite do Joelho/terapia , Osteoartrite do Joelho/complicações , Feminino , Masculino , Pessoa de Meia-Idade , Terapia por Exercício/métodos , Idoso , Resultado do Tratamento , Amplitude de Movimento Articular , Terapia Combinada , Avaliação da Deficiência , Terapia por Estimulação ElétricaRESUMO
Resumen Objetivo: Comprar el dolor en pacientes posoperados de artroscopia de rodilla después de ser sometidos a tratamiento con láser terapéutico versus corrientes TENS. Materiales y método: Se realizó un estudio de tipo experimental, longitudinal, prospectivo, y comparativo que incluyó a 14 pacientes que acudieron al área de Medicina Física y Rehabilitación del Hospital Central Militar, identificados con el diagnóstico de posoperados de artroscopia de rodilla, mediante aleatorización simple, por sorteo, para incluir a los pacientes en los grupos de trabajo: tratamiento con láser terapéutico versus tratamiento con corrientes TENS. Se aplicó la escala visual análoga en la evaluación, y se realizó la medición de ambas rodillas por circometria, pre y postratamiento para medir el edema. El análisis se realizó con el programa estadístico SPSS versión 26 y se utilizó la prueba T de student. Nivel de significancia p<0.05. Resultados: Ingresaron al estudio un total de 14 pacientes, a 7 se les dio en tratamiento con láser terapéutico y a 7 con corrientes TENS. El 50% (7 pacientes) fueron mujeres y el 50% (7 pacientes) fueron hombres. La edad promedio fue de 44.85 años. Al analizar la escala visual análoga, en la evaluación pre y postratamiento ambos grupos disminuyeron su intensidad de dolor con diferencias estadísticamente significativas, (con una desviación típica de 1.15 para TENS y una desviación típica de 1.35) al comparar ambos resultados, se observó una diferencia estadisticamente significativa a favor de las corrientes TENS sobre el láser terapeutico p<0.0001. Conclusiones: En ambos tratamientos, se observaron mejorías en el dolor percibido por los pacientes. Teniendo mayor disminución del dolor con las corrientes TENS. Por lo tanto, en este estudio se llega a la conclusión que este tratamiento es más efectivo que el láser terapéutico en el manejo del dolor en pacientes posoperados de artroscopia de rodilla. No se observó mejoría en el edema en ninguno de los tratamientos.
Abstract Objective: To measure pain in postoperative knee arthroscopy patients after undergoing treatment with therapeutic laser vs. TENS currents. Materials and method: An experimental, longitudinal, prospective, and comparative study was carried out that included 14 patients who attended the Physical Medicine and Rehabilitation area of the Hospital Central Militar, identified with the diagnosis of postoperative knee arthroscopy, through randomization. simple, by lottery, to include patients in the work groups: treatment with therapeutic laser vs treatment with TENS currents. The visual analogue scale was applied in the evaluation, and measurement of both knees was carried out by circometry, pre- and post-treatment to measure edema. The analysis was carried out with the statistical program SPSS version 26 and the student's T test was used. Significance level p<0.05. Results: A total of 14 patients entered the study, 7 were treated with therapeutic laser and 7 with TENS currents. 50% (7 patients) were women and 50% (7 patients) were men. The average age was 44.85 years. When analyzing the visual analog scale, in the pre- and post-treatment evaluation, both groups decreased their pain intensity with statistically significant differences (with a standard deviation of 1.15 for TENS and a standard deviation of 1.35). When comparing both results, a difference was observed. statistically significant difference in favor of TENS Currents over therapeutic laser p<0.0001. Conclusions: In both treatments, both the therapeutic laser and the TENS currents, improvements were observed in the pain perceived by the patients. Having a greater reduction in pain with TENS currents. Therefore, in this study we conclude that treatment with TENS currents is more effective than therapeutic laser in pain management in post-knee arthroscopy patients. No improvement in edema was observed in any of the treatments.
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This study comprehensively analyzes the magnetically induced current density of polycyclic compounds labeled as "aromatic chameleons" since they can arrange their π-electrons to exhibit aromaticity in both the ground and the lowest triplet state. These compounds comprise benzenoid moieties fused to a central skeleton with 4n π-electrons and traditional magnetic descriptors are biased due to the superposition of local magnetic responses. In the S0 state, our analysis reveals that the molecular constituent fragments preserve their (anti)aromatic features in agreement with two types of resonant structures: one associated with aromatic benzenoids and the other with a central antiaromatic ring. Regarding the T1 state, a global and diatropic ring current is revealed. Our aromaticity study is complemented with advanced electronic and geometric descriptors to consider different aspects of aromaticity, particularly important in the evaluation of excited state aromaticity. Remarkably, these descriptors consistently align with the general features on the main delocalization pathways in polycyclic hydrocarbons consisting of fused 4n π-electron rings. Moreover, our study demonstrates an inverse correlation between the singlet-triplet energy difference and the antiaromatic character of the central ring in S0.
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Pelagic Sargassum in the Gulf of Mexico (GoM) plays an important role in ocean biology and ecology, yet our knowledge of its origins and transport pathways is limited. Here, using satellite observations of Sargassum areal density and ocean surface currents between 2000 and 2023, we show that large amounts of Sargassum in the GoM can either originate from the northwestern GoM or be a result of physical transport from the northwestern Caribbean Sea, both with specific transport pathways. Sargassum of the northwestern GoM can be transported to the eastern GoM by ocean currents and eddies, eventually entering the Sargasso Sea. Sargassum entering the GoM from the northwestern Caribbean Sea can be transported in three different directions, with the northward and eastward transports governed by the Loop Current System (LCS) and westward transport driven by the westward extension of the LCS, the propagation or relaying of ocean eddies, the wind-driven westward currents on the Campeche Bank with or without eddies, and the westward currents with/without currents associated with eddies in the northern/central GoM. Overall, the spatial distribution patterns of pelagic Sargassum in the GoM are strongly influenced by the LCS and relevant eddies.
Assuntos
Sargassum , Golfo do México , Meio Ambiente , Região do Caribe , EcologiaRESUMO
Nowadays, an active research topic is the connection between Clar's rule, aromaticity, and magnetic properties of polycyclic benzenoid hydrocarbons. In the present work, we employ a meticulous magnetically induced current density analysis to define the net current flowing through any cyclic circuit, connecting it to aromaticity based on the ring current concept. Our investigation reveals that the analyzed polycyclic systems display a prominent global ring current, contrasting with subdued semi-local and local ring currents. These patterns align with Clar's aromatic π-sextets only in cases where migrating π-sextet structures are invoked. The results of this study will enrich our comprehension of aromaticity and magnetic behavior in such systems, offering significant insights into coexisting ring current circuits in these systems.
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Ouabain, a substance originally obtained from plants, is now classified as a hormone because it is produced endogenously in certain animals, including humans. However, its precise effects on the body remain largely unknown. Previous studies have shown that ouabain can influence the phenotype of epithelial cells by affecting the expression of cell-cell molecular components and voltage-gated potassium channels. In this study, we conducted whole-cell clamp assays to determine whether ouabain affects the activity and/or expression of TRPV4 channels. Our findings indicate that ouabain has a statistically significant effect on the density of TRPV4 currents (dITRPV4), with an EC50 of 1.89 nM. Regarding treatment duration, dITRPV4 reaches its peak at around 1 h, followed by a subsequent decline and then a resurgence after 6 h, suggesting a short-term modulatory effect related to on TRPV4 channel activity and a long-term effect related to the promotion of synthesis of new TRPV4 channel units. The enhancement of dITRPV4 induced by ouabain was significantly lower in cells seeded at low density than in cells in a confluent monolayer, indicating that the action of ouabain depends on intercellular contacts. Furthermore, the fact that U73122 and neomycin suppress the effect caused by ouabain in the short term suggests that the short-term induced enhancement of dITRPV4 is due to the depletion of PIP2 stores. In contrast, the fact that the long-term effect is inhibited by PP2, wortmannin, PD, FR18, and IKK16 suggests that cSrc, PI3K, Erk1/2, and NF-kB are among the components included in the signaling pathways.
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Ouabaína , Canais de Cátion TRPV , Humanos , Animais , Ouabaína/farmacologia , Canais de Cátion TRPV/genética , Canais de Cátion TRPV/metabolismo , Transdução de Sinais , Células Epiteliais/metabolismo , ATPase Trocadora de Sódio-Potássio/metabolismoRESUMO
Background: Long-distance rafting on anthropogenic marine debris (AMD) is thought to have a significant impact on global marine biogeography and the dispersal of non-indigenous species. Therefore, early identification of arrival sites of AMD and its epibionts is crucial for the prioritization of preventive measures. As accumulation patterns along global coastlines are largely unstudied, we tested if existing oceanographic models and knowledge about upstream sources of litter and epibionts can be used as a simple and cost-efficient approach for predicting probable arrival sites of AMD-rafting biota in coastal zones. Methods: Using the Southeast Pacific as a model system, we studied daily accumulation rates, composition, and minimum floating times of AMD with and without epibionts on seven sandy beaches, covering the oceanic environment (Rapa Nui/Easter Island) and three regions (south, centre, north) along the Chilean continental coast, over a minimum of 10 consecutive days, and we contrast our results with predictions from published models. Results: Total AMD accumulation rates varied from 56 ± 36 (mean ± standard deviation) to 388 ± 433 items km-1 d-1 and differed strongly between regions, in accordance with local geomorphology and socioeconomic conditions (presence of larger cities and rivers upstream, main economic activities, etc.). Daily accumulation of items with pelagic epibionts (indicators of a pelagic trajectory) ranged from 46 ± 29 (Rapa Nui) to 0.0 items km-1 d-1 (northern continental region). Minimum floating times of rafts, as estimated from the size of pelagic epibionts, were longest in the South Pacific Subtropical Gyre's (SPSG) centre region, followed by the high-latitude continental region under the influence of the onshore West Wind Drift, and decreased along the continental alongshore upwelling current, towards lower latitudes. Apart from pelagic rafters, a wide range of benthic epibionts, including invasive and cryptogenic species, was found on rafts at the continental beaches. Similarly, we present another record of local benthic corals Pocillopora sp., on Rapa Nui rafts. Discussion: Our results agree with the predictions made by recent models based on the prevailing wind and surface current regimes, with high frequencies of long-distance rafting in the oceanic SPSG centre and very low frequencies along the continental coast. These findings confirm the suitability of such models in predicting arrival hotspots of AMD and rafting species. Moreover, storm surges as well as site-related factors seem to influence AMD arrival patterns along the Chilean continental coast and might cause the observed high variability between sampling sites and days. Our results highlight the possible importance of rafting as a vector of along-shore dispersal and range expansions along the SE Pacific continental coast and add to the discussion about its role in benthic species dispersal between South Pacific oceanic islands.
Assuntos
Antozoários , Monitoramento Ambiental , Animais , Monitoramento Ambiental/métodos , Plásticos , Oceanos e Mares , Biota , PolinésiaRESUMO
Voltage-gated calcium channels (CaV) conduct Ca2+ influx promoting neurotransmitters and hormone release. CaV are finely regulated by voltage-dependent and independent pathways either by G-protein-coupled receptors (GPCRs) or intramembrane lipids, respectively, in neurons and glands. Interestingly, pancreatic ß-cells are abundantly innervated by both sympathetic and parasympathetic neurons, while a variety of high-voltage activated (HVA) Ca2+ channels are present in these cells. Thus, autonomic system seems to exert a tonic inhibition on HVA Ca2+ channels throughout GPCRs, constitutively preventing hormone secretion. Therefore, this work aimed to investigate noradrenergic and cholinergic inhibition of HVA Ca2+ channels in pancreatic ß-cells. Experiments were conducted in pancreatic ß-cells of rat by using patch-clamping methods, immunocytochemistry, pharmacological probes, and biochemical reagents. A voltage-clamp protocol with a strong depolarizing prepulse was used to unmask tonic inhibition. Herein, we consistently find a basal tonic inhibition of HVA Ca2+ channels according to a GPCRs regulation. Facilitation ratio is enhanced by noradrenaline (NA) according to a voltage-dependent regulation and a membrane-delimited mechanism, while no facilitation changes are observed with carbachol or phosphatidylinositol 4,5-bisphosphate (PIP2). Furthermore, carbachol or intramembrane lipids, such as PIP2, do not change facilitation ratio according to a voltage-independent regulation. Together, HVA Ca2+ channels of pancreatic ß-cells are constitutively inhibited by GPCRs, suggesting a natural brake preventing cells from exhaustive insulin secretion.NEW & NOTEWORTHY Our results support the hypothesis that GPCRs tonically inhibit HVA Ca2+ channels in pancreatic ß-cells. A voltage-clamp protocol with a strong depolarizing prepulse was used to unmask voltage-dependent inhibition of Ca2+ channels. The novelty of these results strengthens the critical role of Gßγ's in Ca2+ channel regulation, highlighting kinetic slowing and increased facilitation ratio. Together, HVA Ca2+ channels of pancreatic ß-cells are constitutively inhibited by GPCRs underlying fine-tuning modulation of insulin secretion.
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Canais de Cálcio , Proteínas de Ligação ao GTP , Ratos , Animais , Canais de Cálcio/metabolismo , Carbacol , Proteínas de Ligação ao GTP/metabolismo , Receptores Acoplados a Proteínas G/metabolismo , Hormônios , Lipídeos , Cálcio/metabolismoRESUMO
Background: Conotoxins exhibit great potential as neuropharmacology tools and therapeutic candidates due to their high affinity and specificity for ion channels, neurotransmitter receptors or transporters. The traditional methods to discover new conotoxins are peptide purification from the crude venom or gene amplification from the venom duct. Methods: In this study, a novel O1 superfamily conotoxin Tx6.7 was directly cloned from the genomic DNA of Conus textile using primers corresponding to the conserved intronic sequence and 3' UTR elements. The mature peptide of Tx6.7 (DCHERWDWCPASLLGVIYCCEGLICFIAFCI) was synthesized by solid-phase chemical synthesis and confirmed by mass spectrometry. Results: Patch clamp experiments on rat DRG neurons showed that Tx6.7 inhibited peak calcium currents by 59.29 ± 2.34% and peak potassium currents by 22.33 ± 7.81%. In addition, patch clamp on the ion channel subtypes showed that 10 µM Tx6.7 inhibited 56.61 ± 3.20% of the hCaV1.2 currents, 24.67 ± 0.91% of the hCaV2.2 currents and 7.30 ± 3.38% of the hNaV1.8 currents. Tx6.7 had no significant toxicity to ND7/23 cells and increased the pain threshold from 0.5 to 4 hours in the mouse hot plate assay. Conclusion: Our results suggested that direct cloning of conotoxin sequences from the genomic DNA of cone snails would be an alternative approach to obtaining novel conotoxins. Tx6.7 could be used as a probe tool for ion channel research or a therapeutic candidate for novel drug development.
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To study whether diabetes mellitus (DM) would cause electrophysiological alterations in nodose ganglion (NG) neurons, we used patch clamp and intracellular recording for voltage and current clamp configuration, respectively, on cell bodies of NG from rats with DM. Intracellular microelectrodes recording, according to the waveform of the first derivative of the action potential, revealed three neuronal groups (A0 , Ainf , and Cinf ), which were differently affected. Diabetes only depolarized the resting potential of A0 (from -55 to -44 mV) and Cinf (from -49 to -45 mV) somas. In Ainf neurons, diabetes increased action potential and the after-hyperpolarization durations (from 1.9 and 18 to 2.3 and 32 ms, respectively) and reduced dV/dtdesc (from -63 to -52 V s-1 ). Diabetes reduced the action potential amplitude while increasing the after-hyperpolarization amplitude of Cinf neurons (from 83 and -14 mV to 75 and -16 mV, respectively). Using whole cell patch clamp recording, we observed that diabetes produced an increase in peak amplitude of sodium current density (from -68 to -176 pA pF-1 ) and displacement of steady-state inactivation to more negative values of transmembrane potential only in a group of neurons from diabetic animals (DB2). In the other group (DB1), diabetes did not change this parameter (-58 pA pF-1 ). This change in sodium current did not cause an increase in membrane excitability, probably explainable by the alterations in sodium current kinetics, which are also induced by diabetes. Our data demonstrate that diabetes differently affects membrane properties of different nodose neuron subpopulations, which likely have pathophysiological implications for diabetes mellitus.
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Diabetes Mellitus , Neurônios Aferentes , Ratos , Animais , Neurônios Aferentes/fisiologia , Potenciais da Membrana/fisiologia , Neurônios/fisiologia , Potenciais de Ação/fisiologia , SódioRESUMO
Background: Conotoxins exhibit great potential as neuropharmacology tools and therapeutic candidates due to their high affinity and specificity for ion channels, neurotransmitter receptors or transporters. The traditional methods to discover new conotoxins are peptide purification from the crude venom or gene amplification from the venom duct. Methods: In this study, a novel O1 superfamily conotoxin Tx6.7 was directly cloned from the genomic DNA of Conus textile using primers corresponding to the conserved intronic sequence and 3' UTR elements. The mature peptide of Tx6.7 (DCHERWDW CPASLLGVIYCCEGLICFIAFCI) was synthesized by solid-phase chemical synthesis and confirmed by mass spectrometry. Results: Patch clamp experiments on rat DRG neurons showed that Tx6.7 inhibited peak calcium currents by 59.29 ± 2.34% and peak potassium currents by 22.33 ± 7.81%. In addition, patch clamp on the ion channel subtypes showed that 10 µM Tx6.7 inhibited 56.61 ± 3.20% of the hCaV1.2 currents, 24.67 ± 0.91% of the hCaV2.2 currents and 7.30 ± 3.38% of the hNaV1.8 currents. Tx6.7 had no significant toxicity to ND7/23 cells and increased the pain threshold from 0.5 to 4 hours in the mouse hot plate assay. Conclusion: Our results suggested that direct cloning of conotoxin sequences from the genomic DNA of cone snails would be an alternative approach to obtaining novel conotoxins. Tx6.7 could be used as a probe tool for ion channel research or a therapeutic candidate for novel drug development.(AU)
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Animais , Cálcio/isolamento & purificação , Conotoxinas/genética , Caramujo Conus/químicaRESUMO
The majority of voltage-gated ion channels contain a defined voltage-sensing domain and a pore domain composed of highly conserved amino acid residues that confer electrical excitability via electromechanical coupling. In this sense, the voltage-gated proton channel (Hv1) is a unique protein in that voltage-sensing, proton permeation and pH-dependent modulation involve the same structural region. In fact, these processes synergistically work in concert, and it is difficult to separate them. To investigate the process of Hv1 voltage sensor trapping, we follow voltage-sensor movements directly by leveraging mutations that enable the measurement of Hv1 channel gating currents. We uncover that the process of voltage sensor displacement is due to two driving forces. The first reveals that mutations in the selectivity filter (D160) located in the S1 transmembrane interact with the voltage sensor. More hydrophobic amino acids increase the energy barrier for voltage sensor activation. On the other hand, the effect of positive charges near position 264 promotes the formation of salt bridges between the arginines of the voltage sensor domain, achieving a stable conformation over time. Our results suggest that the activation of the Hv1 voltage sensor is governed by electrostatic-hydrophobic interactions, and S4 arginines, N264 and selectivity filter (D160) are essential in the Ciona-Hv1 to understand the trapping of the voltage sensor.
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Antifibrinolíticos , Ciona , Animais , Prótons , Aminoácidos , ArgininaRESUMO
Synapse unsilencing is an essential mechanism for experience-dependent plasticity. Here, we showed that the application of the ligand Wnt-5a converts glutamatergic silent synapses into functional ones by increasing both α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) and N-methyl-D-aspartate (NMDA) currents (IAMPA and INMDA, respectively). These effects were mimicked by the hexapeptide Foxy-5 and inhibited by secreted frizzled-related protein sFRP-2. INMDA potentiation was produced by increased synaptic potency, followed by an increase in the probability of release (Pr), even in the presence of 7-nitro-2,3-dioxo-1,4-dihydroquinoxaline-6-carbonitrile (CNQX). At a longer time of Wnt-5a exposure, the Pr increments were higher in INMDA than in IAMPA. In the presence of NMDAR inhibitors, Wnt-5a-induced conversion was fully inhibited in 69.0% of silent synapses, whereas in the remaining synapses were converted into functional one. Our study findings showed that the Wnt-5a-activated pathway triggers AMPAR insertion into mammalian glutamatergic synapses, unsilencing non-functional synapses and promoting the formation of nascent synapses during the early postnatal development of the brain circuits.
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A low-lying structure is revealed for the CuB12 - cluster, which is bowl-shaped. It consists of a triangular CuB2 base and a B10 rim. Molecular dynamics simulations indicates its structural robustness; at an elevated temperature (600â K), the base rotates reversibly within the B10 perimeter. Chemical bonding analysis detects 2σ- and 3π-delocalized bonds, suggesting double aromaticity. This is also confirmed by two diatropic and concentric ring currents under an external magnetic field.
RESUMO
The epidemiological association between bacterial or viral maternal infections during pregnancy and increased risk for developing psychiatric disorders in offspring is well documented. Numerous rodent and non-human primate studies of viral- or, to a lesser extent, bacterial-induced maternal immune activation (MIA) have documented a series of neurological alterations that may contribute to understanding the pathophysiology of schizophrenia and autism spectrum disorders. Long-term neuronal and behavioral alterations are now ascribed to the effect of maternal proinflammatory cytokines rather than the infection itself. However, detailed electrophysiological alterations in brain areas relevant to psychiatric disorders, such as the dorsal hippocampus, are lacking in response to bacterial-induced MIA. This study determined if electrophysiological and morphological alterations converge in CA1 pyramidal cells (CA1 PC) from the dorsal hippocampus in bacterial-induced MIA offspring. A series of changes in the functional expression of K+ and Na+ ion channels altered the passive and active membrane properties and triggered hyperexcitability of CA1 PC. Contributing to the hyperexcitability, the somatic A-type potassium current (IA) was decreased in MIA CA1 PC. Likewise, the spontaneous glutamatergic and GABAergic inputs were dysregulated and biased toward increased excitation, thereby reshaping the excitation-inhibition balance. Consistent with these findings, the dendritic branching complexity of MIA CA1 PC was reduced. Together, these morphophysiological alterations modify CA1 PC computational capabilities and contribute to explaining cellular alterations that may underlie the cognitive symptoms of MIA-associated psychiatric disorders.
Assuntos
Imunidade , Neurônios , Canais de Potássio , Animais , Transtorno do Espectro Autista/imunologia , Região CA1 Hipocampal/citologia , Regulação para Baixo , Feminino , Neurônios/metabolismo , Canais de Potássio/metabolismo , Gravidez , Células Piramidais/imunologia , Esquizofrenia/imunologiaRESUMO
The leakage current (LC) caused by the surface contamination of insulators, together with environmental variables, is one of the most basic online monitoring parameters for insulator status. However, the impact of weather conditions such as temperature, air humidity, and dew point on the LC has not been deeply studied until now. In this paper, based on meteorological data obtained online and LC obtained with an optical fiber sensor, installed in 500-kV insulator strings of a transmission line, the impact of weather conditions was studied. Results indicate that the LCs follow a specific pattern, according to weather conditions. The system has been continuously monitoring LC, humidity, temperature, and dew point uninterrupted for three years, sending the acquired data to a web page; therefore, it has been demonstrated to be robust, reliable, and repetitive. The sensor features the broadband response and acquisition capabilities of partial discharge pulses in high-voltage insulators, allowing the detection of high-frequency pulses. When comparing the LC measured in this work with those from other works, our measurements are substantially higher; this is due to the type of pollution found in this specific situation, which includes iron oxide powder, producing a conductive layer over the insulator surface that, unlike sea salt, does not depend on humidity to conduct an LC. One of the conclusions reached in this work is that partial discharge surges are caused when the local temperature reaches the dew point and not simply from the presence of high humidity, as stated in many works dealing with LCs. The monitored LC can be used as an indicative parameter of a possible flashover, enabling the proper planning of insulator predictive maintenance, either by jet-washing the surface or even changing the insulators when they are damaged.