RESUMO
Voltage-gated Na+ channels are crucial to action potential propagation in excitable tissues. Because of the high amplitude and rapid activation of the Na+ current, voltage-clamp measurements are very challenging and are usually performed at room temperature. In this study, we measured Na+ current voltage-dependence in stem cell-derived cardiomyocytes at physiological temperature. While the apparent activation and inactivation curves, measured as the dependence of current amplitude on voltage, fall within the range reported in previous studies, we identified a systematic error in our measurements. This error is caused by the deviation of the membrane potential from the command potential of the amplifier. We demonstrate that it is possible to account for this artifact using computer simulation of the patch-clamp experiment. We obtained surprising results through patch-clamp model optimization: a half-activation of -11.5 mV and a half-inactivation of -87 mV. Although the half-activation deviates from previous research, we demonstrate that this estimate reproduces the conduction velocity dependence on extracellular potassium concentration. KEY POINTS: Voltage-gated Na+ currents play a crucial role in excitable tissues including neurons, cardiac and skeletal muscle. Measurement of Na+ current is challenging because of its high amplitude and rapid kinetics, especially at physiological temperature. We have used the patch-clamp technique to measure human Na+ current voltage-dependence in human induced pluripotent stem cell-derived cardiomyocytes. The patch-clamp data were processed by optimization of the model accounting for voltage-clamp experiment artifacts, revealing a large difference between apparent parameters of Na+ current and the results of the optimization. We conclude that actual Na+ current activation is extremely depolarized in comparison to previous studies. The new Na+ current model provides a better understanding of action potential propagation; we demonstrate that it explains propagation in hyperkalaemic conditions.
Assuntos
Células-Tronco Pluripotentes Induzidas , Sódio , Humanos , Simulação por Computador , Sódio/fisiologia , Temperatura , Miócitos Cardíacos , Modelos TeóricosRESUMO
In addition to the hallmark muscle stiffness, patients with recessive myotonia congenita (Becker disease) experience debilitating bouts of transient weakness that remain poorly understood despite years of study. We performed intracellular recordings from muscle of both genetic and pharmacologic mouse models of Becker disease to identify the mechanism underlying transient weakness. Our recordings reveal transient depolarizations (plateau potentials) of the membrane potential to -25 to -35 mV in the genetic and pharmacologic models of Becker disease. Both Na+ and Ca2+ currents contribute to plateau potentials. Na+ persistent inward current (NaPIC) through NaV1.4 channels is the key trigger of plateau potentials and current through CaV1.1 Ca2+ channels contributes to the duration of the plateau. Inhibiting NaPIC with ranolazine prevents the development of plateau potentials and eliminates transient weakness in vivo. These data suggest that targeting NaPIC may be an effective treatment to prevent transient weakness in myotonia congenita.
Myotonia is a neuromuscular condition that causes problems with the relaxation of muscles following voluntary movements. One type of myotonia is Becker disease, also called recessive myotonia congenita. This is a genetic condition that causes muscle stiffness as a result of involuntary muscle activity. Patients may also suffer transient weakness for a few seconds or as long as several minutes after initiating a movement. The cause of these bouts of temporary weakness is still unclear, but there are hints that it could be linked to the muscle losing its excitability, the ability to respond to the stimuli that make it contract. However, this is at odds with findings that show that muscles in Becker disease are hyperexcitable. Muscle excitability depends on the presence of different concentrations of charged ions (positively charged sodium, calcium and potassium ions and negatively charged chloride ions) inside and outside of each muscle cells. These different concentrations of ions create an electric potential across the cell membrane, also called the 'membrane potential'. When a muscle cell gets stimulated, proteins on the cell membrane known as ion channels open. This allows the flow of ions between the inside and the outside of the cell, which causes an electrical current that triggers muscle contraction. To better understand the causes behind this muscle weakness, Myers et al. used mice that had either been genetically manipulated or given drugs to mimic Becker disease. By measuring both muscle force and the electrical currents that drive contraction, Myers et al. found that the mechanism underlying post-movement weakness involved a transient change in the concentrations of positively charged ions inside and outside the cells. Further experiments showed that proteins that regulate the passage of both sodium and calcium in and out of the cell called sodium and calcium channels contributed to this change in concentration. In addition, Myers et al. discovered that using a drug called ranolazine to stop sodium ions from entering the cell eliminated transient weakness in live mice. These findings suggest that in Becker disease, muscles cycle rapidly between being hyperexcited or not able to be excited, and that targeting the flow of sodium ions into the cell could be an effective treatment to prevent transient weakness in myotonia congenita. This study paves the way towards the development of new therapies to treat Becker disease as well as other muscle ion channel diseases with transient weakness such as periodic paralysis.
Assuntos
Potenciais da Membrana/fisiologia , Miotonia Congênita/fisiopatologia , Animais , Modelos Animais de Doenças , Feminino , Masculino , Camundongos , Miotonia Congênita/diagnóstico , Miotonia Congênita/genética , Sódio/fisiologiaRESUMO
OBJECTIVES: To investigate the role of sodium in intravesical absorption of water in the bladder and the sodium pathway in the urothelium. METHODS: Adult female Sprague-Dawley rats received either saline or a 5% glucose solution injection into their bladders. The changes in intravesical fluid volume; concentrations of sodium and chlorine and osmolality; and expression of aquaporin-2, epithelial sodium channel, and claudins were compared after 3 hours. RESULTS: Intravesical volume decreased significantly in the saline group compared to that in the 5% glucose solution group. The expression of claudin-3 and -6 was higher in the saline group than in the glucose group. There was a significant correlation between changes in the intravesical saline volume and the concentration of sodium and chlorine. Intravesical administration of amiloride did not affect changes in the fluid volume and concentration of sodium. CONCLUSIONS: The presence of sodium is important for the absorption of intravesical fluid through aquaporin-2 in the urinary bladders of rats. Claudin-3 and -6 may be associated with the transport of sodium through the bladder urothelium.
Assuntos
Sódio/fisiologia , Bexiga Urinária/fisiologia , Urotélio/fisiologia , Administração Intravesical , Animais , Aquaporina 2/metabolismo , Cloro/metabolismo , Claudinas/metabolismo , Feminino , Concentração Osmolar , Ratos , Ratos Sprague-Dawley , Sódio/administração & dosagem , Sódio/metabolismo , Sódio/farmacologia , Canais de Sódio/fisiologia , Bexiga Urinária/metabolismo , Urina/química , Urotélio/metabolismoRESUMO
ETHNOPHARMACOLOGY RELEVANCE: Pain is an unpleasant sensory and emotional experience, often accompanied by the occurrence of a variety of diseases. More than 800 kinds of traditional Chinese medicines (TCM) has now been reported for pain relief and several monomers have been developed into novel analgesic drugs. Bupleurum chinense and Angelica biserrata were representatives of the TCM that are currently available for the treatment of pain. AIM OF THE STUDY: The study aims to detect the potential analgesic activity of each monomer of Bupleurum chinense and Angelica biserrata and to explore whether Nav1.7 is one of the targets for its analgesic activity. MATERIALS AND METHODS: In this study, five monomers from Bupleurum chinense (Saikosaponin A, Saikosaponin B1, Saikosaponin B2, Saikosaponin C, Saikosaponin D) and five monomers from the Angelica biserrata (Osthole, Xanthotoxin, Imperatorin, Isoimperatorin, Psoralen) were examined by whole-cell patch-clamp on Nav1.7, which was closely associated with pain. Classical mouse pain models were also used to further verify the analgesic activity in vivo. RESULTS: The results showed that monomers of Saikosaponins and Angelica biserrata all inhibited the peak currents of Nav1.7, indicating that Nav1.7 might be involved in the analgesic mechanism of Saikosaponins and Angelica biserrata. Among them, Saikosaponin A and Imperatorin showed the strongest inhibitory effect on Nav1.7. Furthermore, both Saikosaponin A and Imperatorin showed inhibitory effects on thermal pain and formalin-induced pain in phase II in vivo. CONCLUSION: The results provide valuable information for future studies on the potential of TCM in alleviating pain.
Assuntos
Analgésicos/farmacologia , Angelica/química , Bupleurum/química , Medicamentos de Ervas Chinesas/farmacologia , Canal de Sódio Disparado por Voltagem NAV1.7/efeitos dos fármacos , Dor/tratamento farmacológico , Analgésicos/química , Analgésicos/uso terapêutico , Animais , Células CHO , Cricetulus , Modelos Animais de Doenças , Medicamentos de Ervas Chinesas/química , Medicamentos de Ervas Chinesas/uso terapêutico , Feminino , Formaldeído/toxicidade , Furocumarinas/farmacologia , Furocumarinas/uso terapêutico , Temperatura Alta/efeitos adversos , Masculino , Medicina Tradicional Chinesa , Camundongos , Ácido Oleanólico/análogos & derivados , Ácido Oleanólico/farmacologia , Ácido Oleanólico/uso terapêutico , Dor/etiologia , Raízes de Plantas/química , Saponinas/farmacologia , Saponinas/uso terapêutico , Sódio/fisiologiaRESUMO
The importance of the intracellular Ca2+ concentration ([Ca2+]i) in neutrophil function has been intensely studied. However, the role of the intracellular Na+ concentration ([Na+]i) which is closely linked to the intracellular Ca2+ regulation has been largely overlooked. The [Na+]i is regulated by Na+ transport proteins such as the Na+/Ca2+-exchanger (NCX1), Na+/K+-ATPase, and Na+-permeable, transient receptor potential melastatin 2 (TRPM2) channel. Stimulating with either N-formylmethionine-leucyl-phenylalanine (fMLF) or complement protein C5a causes distinct changes of the [Na+]i. fMLF induces a sustained increase of [Na+]i, surprisingly, reaching higher values in TRPM2-/- neutrophils. This outcome is unexpected and remains unexplained. In both genotypes, C5a elicits only a transient rise of the [Na+]i. The difference in [Na+]i measured at t = 10 min after stimulation is inversely related to neutrophil chemotaxis. Neutrophil chemotaxis is more efficient in C5a than in an fMLF gradient. Moreover, lowering the extracellular Na+ concentration from 140 to 72 mM improves chemotaxis of WT but not of TRPM2-/- neutrophils. Increasing the [Na+]i by inhibiting the Na+/K+-ATPase results in disrupted chemotaxis. This is most likely due to the impact of the altered Na+ homeostasis and presumably NCX1 function whose expression was shown by means of qPCR and which critically relies on proper extra- to intracellular Na+ concentration gradients. Increasing the [Na+]i by a few mmol/l may suffice to switch its transport mode from forward (Ca2+-efflux) to reverse (Ca2+-influx) mode. The role of NCX1 in neutrophil chemotaxis is corroborated by its blocker, which also causes a complete inhibition of chemotaxis.
Assuntos
Quimiotaxia de Leucócito/imunologia , Homeostase/imunologia , Sódio/fisiologia , Canais de Cátion TRPM/fisiologia , Animais , Cálcio/fisiologia , Linhagem Celular Tumoral , Células Cultivadas , Quimiotaxia de Leucócito/efeitos dos fármacos , Complemento C5a/imunologia , Complemento C5a/farmacologia , Líquido Intracelular/imunologia , Leucemia Mieloide , Camundongos , Camundongos Endogâmicos C57BL , N-Formilmetionina Leucil-Fenilalanina/farmacologia , Ativação de Neutrófilo/efeitos dos fármacos , Trocador de Sódio e Cálcio/fisiologia , ATPase Trocadora de Sódio-Potássio/fisiologia , Canais de Cátion TRPM/deficiênciaRESUMO
We found previously that intravenous kynurenic acid (KYNA), a native broad spectrum glutamate antagonist, increases renal blood flow and induces natriuresis in anesthetized spontaneously hypertensive rats (SHR). Since such changes may affect systemic circulation and can potentially find therapeutic application, in this study we examined long term influence of orally administered KYNA on systemic and renal hemodynamics and renal excretion in conscious SHR. KYNA was administered in drinking water at a dose of 25 mg/kg/day for 3 weeks. Heart rate (HR), systolic (SBP), and mean arterial pressure (MAP) were measured through telemetry. The records were taken at the beginning of the study (control, day 0), and then on day 7, 14, and 21 of treatment. Diuresis (V), total solute excretion (UosmV), and sodium excretion (UNaV) were determined on days 0, 7, and 14. KYNA consistently decreased HR, from 319 ± 8 to 291 ± 5, 299 ± 9 and 284 ± 6 beats/min on day 7, 14, and 21, respectively, (- 9, - 6, and - 11%; p < 0.01-0.0001); HR was stable in the solvent group. SBP, MAP, V, and UNaV were not affected by KYNA, whereas UosmV increased modestly. Chronic oral administration of KYNA to conscious SHR decreased HR without affecting MAP. Since tachycardia is an independent risk factor for cardiovascular disorders, and most drugs used to decrease HR have strong inotropic negative or hypotensive effect, such selective action seems of therapeutic potential. Moreover, food supplementation with KYNA can be considered in the prevention of heart diseases.
Assuntos
Frequência Cardíaca/efeitos dos fármacos , Hipertensão/fisiopatologia , Ácido Cinurênico/farmacologia , Animais , Pressão Arterial/efeitos dos fármacos , Rim/efeitos dos fármacos , Rim/patologia , Rim/fisiologia , Masculino , Miocárdio/patologia , Ratos Endogâmicos SHR , Sódio/fisiologiaRESUMO
OBJECTIVES: (1) To detect the whether the effects of simulated ischemia on INa of rat left ventricular myocytes in a time-dependent manner and the effects of atorvastatin on ischemia INa; (2) To investigate the effects of atorvastatin on INa of rat-simulated ischemia/reperfusion (I/R) ventricular cells. MATERIALS AND METHODS: Ventricular cells were enzymatically isolated by Langendorff perfusion system. Whole-cell patch clamp was applied to detect INa level. Some elements of extracellular fluid were hanged to simulate the status of normal, I and R condition. Then the effects of atorvastatin on INa were observed. RESULTS: (1) During simulated reperfusion, INa decreased and atorvastatin further suppressed the reduction degree. (2) At test potential -40 mV, no difference was detected among peak INa amplitude of ischemia for 20 min, reperfusion phase 3/5/7/9 min in continuous ischemia (I) group (p = 0.275). In I/R group, peak INa amplitude continuously decreased at 3 min (p = 0.005) and 9 min (p = 0.041). In atorvastatin intervention + I/R (Statin + I/R) group, peak INa amplitude at reperfusion 3 min decreased compared with ischemia phase (p = 0.000), while no significant difference was detected between 3 and 9 min (p = 0.858). The differences were significant at the same time point between groups. At reperfusion 3/5/7/9 min, peak INa of the I/R group was lower than the ischemia group (all p = 0.000), same as the Statin + I/R group (p = 0.000, p = 0.003, p = 0.006, and p = 0.001). Peak INa of the Statin + I/R group was higher than the I/R group at the same time point (p = 0.011, p = 0.033, p = 0.003, p = 0.003). There was no change in the I group during reperfusion phase (p > 0.05). In I/R group, V1/2 (mV) shifted from -58.87 ± 3.36 to -54.33 ± 2.40, k (mV) shifted from 1.25 ± 0.59 to 1.91 ± 0.84 (p < 0.05). In the Statin + I/R group, V1/2 (mV) increased from -57.80 ± 2.97 to -52.76 ± 3.14 (p < 0.01), no change was observed in k (p > 0.05). CONCLUSIONS: (1) In the status of reperfusion, INa decreased more than that in the status of ischemia. (2) Atorvastatin protected the cells from reduction of INa during long-time simulated (>15 min) I/R. (3) Overall, atorvastatin affected INa of the normal, simulated ischemic/reperfusion cell in rat left ventricle by blocking sodium channel -directly.
Assuntos
Antiarrítmicos/farmacologia , Atorvastatina/farmacologia , Ventrículos do Coração/fisiopatologia , Isquemia Miocárdica/fisiopatologia , Traumatismo por Reperfusão Miocárdica/fisiopatologia , Canais de Sódio/efeitos dos fármacos , Animais , Antiarrítmicos/uso terapêutico , Atorvastatina/uso terapêutico , Feminino , Ventrículos do Coração/efeitos dos fármacos , Ventrículos do Coração/metabolismo , Masculino , Isquemia Miocárdica/tratamento farmacológico , Isquemia Miocárdica/metabolismo , Traumatismo por Reperfusão Miocárdica/tratamento farmacológico , Traumatismo por Reperfusão Miocárdica/patologia , Miócitos Cardíacos/efeitos dos fármacos , Distribuição Aleatória , Ratos , Ratos Wistar , Sódio/fisiologia , Canais de Sódio/fisiologia , Fatores de TempoRESUMO
KEY POINTS: Employing quantitative Na+ -imaging and Förster resonance energy transfer-based imaging with ATeam1.03YEMK (ATeam), we studied the relation between activity-induced Na+ influx and intracellular ATP in CA1 pyramidal neurons of the mouse hippocampus. Calibration of ATeam in situ enabled a quantitative estimate of changes in intracellular ATP concentrations. Different paradigms of stimulation that induced global Na+ influx into the entire neuron resulted in decreases in [ATP] in the range of 0.1-0.6 mm in somata and dendrites, while Na+ influx that was locally restricted to parts of dendrites did not evoke a detectable change in dendritic [ATP]. Our data suggest that global Na+ transients require global cellular activation of the Na+ /K+ -ATPase resulting in a consumption of ATP that transiently overrides its production. For recovery from locally restricted Na+ influx, ATP production as well as fast intracellular diffusion of ATP and Na+ might prevent a local drop in [ATP]. ABSTRACT: Excitatory neuronal activity results in the influx of Na+ through voltage- and ligand-gated channels. Recovery from accompanying increases in intracellular Na+ concentrations ([Na+ ]i ) is mainly mediated by the Na+ /K+ -ATPase (NKA) and is one of the major energy-consuming processes in the brain. Here, we analysed the relation between different patterns of activity-induced [Na+ ]i signalling and ATP in mouse hippocampal CA1 pyramidal neurons by Na+ imaging with sodium-binding benzofurane isophthalate (SBFI) and employing the genetically encoded nanosensor ATeam1.03YEMK (ATeam). In situ calibrations demonstrated a sigmoidal dependence of the ATeam Förster resonance energy transfer ratio on the intracellular ATP concentration ([ATP]i ) with an apparent KD of 2.6 mm, indicating its suitability for [ATP]i measurement. Induction of recurrent network activity resulted in global [Na+ ]i oscillations with amplitudes of â¼10 mm, encompassing somata and dendrites. These were accompanied by a steady decline in [ATP]i by 0.3-0.4 mm in both compartments. Global [Na+ ]i transients, induced by afferent fibre stimulation or bath application of glutamate, caused delayed, transient decreases in [ATP]i as well. Brief focal glutamate application that evoked transient local Na+ influx into a dendrite, however, did not result in a measurable reduction in [ATP]i . Our results suggest that ATP consumption by the NKA following global [Na+ ]i transients temporarily overrides its availability, causing a decrease in [ATP]i . Locally restricted Na+ transients, however, do not result in detectable changes in local [ATP]i , suggesting that ATP production, together with rapid intracellular diffusion of both ATP and Na+ from and to unstimulated neighbouring regions, counteracts a local energy shortage under these conditions.
Assuntos
Trifosfato de Adenosina/fisiologia , Hipocampo/fisiologia , Células Piramidais/fisiologia , Sódio/fisiologia , Animais , Feminino , Masculino , Camundongos Endogâmicos BALB C , Camundongos TransgênicosRESUMO
Sodium intake is undoubtedly indispensable for normal body functions but can be detrimental when taken in excess of dietary requirements. The consequences of excessive salt intake are becoming increasingly clear as high salt consumption persists across the globe. Salt has long been suspected to promote the development of hypertension and cardiovascular diseases and is now also recognized as a potential modulator of inflammatory and autoimmune diseases through its direct and indirect effects on immune cells. The finding that, in addition to the kidneys, other organs such as the skin regulate sodium levels in the body prompted new hypotheses, including the concept that skin-resident macrophages might participate in tissue sodium regulation through their interactions with lymphatic vessels. Moreover, immune cells such as macrophages and different T cell subsets are found in sodium-rich interstitial microenvironments, where sodium levels modulate their function. Alterations to the intestinal bacterial community induced by excess dietary salt represent another relevant axis whereby salt indirectly modulates immune cell function. Depending on the inflammatory context, sodium might either contribute to protective immunity (for example, by enhancing host responses against cutaneous pathogens) or it might contribute to immune dysregulation and promote the development of cardiovascular and autoimmune diseases.
Assuntos
Imunidade Celular/fisiologia , Sódio/fisiologia , Animais , Humanos , Hipertensão/etiologia , Hipertensão/imunologia , Imunidade/fisiologia , Sódio/metabolismo , Linfócitos T/metabolismo , Linfócitos T/fisiologiaRESUMO
Orexins/hypocretins are hypothalamic neuropeptides that have a variety of functions, including maintenance of arousal, control over the sleep/wake cycle, reward and feeding. Accumulating evidence links orexins to the time-keeping system with a documented action in the master clock-the suprachiasmatic nucleus. The intergeniculate leaflet (IGL) is a thalamic structure with the well-known function of collecting photic and non-photic cues to adjust the rhythm of the suprachiasmatic nucleus to changing environmental conditions. The IGL consists of GABAergic neurons that are intrinsically active, even in slice preparations. Our previous studies revealed the excitatory postsynaptic effects of orexins on single IGL neurons, even though the ionic mechanism underlying this effect remained elusive. Therefore, in this study, we used patch clamp electrophysiology to identify the ions and distinct ion channels responsible for the observed depolarisations. The major finding of this article is that the orexin A-evoked depolarisation of IGL neurons depends on non-selective cation channels, implicating the orexinergic tone in establishing the basal firing rate in these cells. The data presented here strengthen the mutual connections between the time-keeping and orexinergic systems.
Assuntos
Corpos Geniculados/efeitos dos fármacos , Canais Iônicos/efeitos dos fármacos , Neurônios/efeitos dos fármacos , Orexinas/farmacologia , Núcleos Talâmicos/efeitos dos fármacos , Animais , Fenômenos Eletrofisiológicos/efeitos dos fármacos , Potenciais Pós-Sinápticos Excitadores/efeitos dos fármacos , Corpos Geniculados/citologia , Masculino , Técnicas de Patch-Clamp , Potássio/fisiologia , Ratos , Ratos Wistar , Sódio/fisiologia , Núcleo Supraquiasmático/efeitos dos fármacos , Núcleos Talâmicos/citologia , Ácido gama-Aminobutírico/fisiologiaRESUMO
IMPACT STATEMENT: This review summarizes the work on transcellular intestinal phosphate absorption, arguing why this pathway is not the predominant pathway in humans consuming a "Western" diet. We then highlight the recent evidence which is strongly consistent with paracellular intestinal phosphate absorption mediating the bulk of intestinal phosphate absorption in humans.
Assuntos
Absorção Intestinal/fisiologia , Fosfatos/farmacocinética , Fósforo na Dieta/farmacocinética , Animais , Ensaios Clínicos como Assunto , Dieta Ocidental , Eletroquímica , Enterócitos/efeitos dos fármacos , Enterócitos/metabolismo , Humanos , Hiperfosfatemia/etiologia , Hiperfosfatemia/fisiopatologia , Hiperfosfatemia/terapia , Absorção Intestinal/efeitos dos fármacos , Isoquinolinas/uso terapêutico , Camundongos , Camundongos Knockout , Niacinamida/uso terapêutico , Compostos Orgânicos/química , Fosfatos/urina , Coelhos , Ratos , Proteínas Recombinantes/metabolismo , Insuficiência Renal Crônica/complicações , Insuficiência Renal Crônica/fisiopatologia , Sódio/fisiologia , Trocador 3 de Sódio-Hidrogênio/antagonistas & inibidores , Trocador 3 de Sódio-Hidrogênio/deficiência , Trocador 3 de Sódio-Hidrogênio/fisiologia , Proteínas Cotransportadoras de Sódio-Fosfato Tipo III/metabolismo , Proteínas Cotransportadoras de Sódio-Fosfato Tipo IIb/antagonistas & inibidores , Proteínas Cotransportadoras de Sódio-Fosfato Tipo IIb/deficiência , Proteínas Cotransportadoras de Sódio-Fosfato Tipo IIb/fisiologia , Sulfonamidas/uso terapêuticoRESUMO
Biomarkers and two clinical rating scales-the Japanese mitochondrial disease-rating scale (JMDRS) and Newcastle mitochondrial disease adult scale (NMDAS)-are clinically used when treating patients with mitochondrial disease. We explored the biomarker(s) and clinical rating scale(s) that are appropriate in preparing the protocol for a future clinical trial of sodium pyruvate (SP) therapy. A 48-week, prospective, single-centre, exploratory, clinical study enrolled 11 Japanese adult patients with genetically, biochemically, and clinically confirmed mitochondrial disease; they had intractable lactic acidosis and received SP (0.5â¯g/kgâ¯t.i.d. PO). Plasma concentrations of lactate and pyruvate, lateral ventricular levels of lactate, and serum concentrations of growth differentiation factor 15 (GDF15) and fibroblast growth factor 21 were measured at baseline and at weeks 12 and 48 of SP therapy. At week 48, plasma lactate (Pâ¯=â¯.004), the lactate/pyruvate ratio (Pâ¯=â¯.012), serum GDF15 (Pâ¯=â¯.020), and lateral ventricular lactate (Pâ¯=â¯.038) decreased significantly from the baseline values; the JMDRS and NMDAS scores did not decrease significantly, although the NMDAS overall score showed a strong tendency (Pâ¯=â¯.059). Two patients with end-stage MELAS at baseline died during SP therapy. The present study showed significant decreases in plasma and lateral ventricular lactate, the L/P ratio, and serum GDF15. Therefore, the protocol for a future clinical study of SP therapy in this patient population needs to include plasma and lateral ventricular lactate, the L/P ratio, and serum GDF15 as diagnostic indicators, and exclude patients with end-stage mitochondrial disease.
Assuntos
Biomarcadores/metabolismo , Mitocôndrias/efeitos dos fármacos , Doenças Mitocondriais/tratamento farmacológico , Doenças Mitocondriais/metabolismo , Ácido Pirúvico/farmacologia , Sódio/fisiologia , Acidose Láctica/tratamento farmacológico , Acidose Láctica/metabolismo , Adolescente , Adulto , Feminino , Fatores de Crescimento de Fibroblastos/metabolismo , Fator 15 de Diferenciação de Crescimento/metabolismo , Humanos , Ácido Láctico/metabolismo , Síndrome MELAS/tratamento farmacológico , Síndrome MELAS/metabolismo , Masculino , Pessoa de Meia-Idade , Mitocôndrias/metabolismo , Estudos Prospectivos , Adulto JovemRESUMO
Mild traumatic injury can modify the key sodium (Na+) current underlying the excitability of neurons. It causes the activation and inactivation properties of this current to become shifted to more negative trans-membrane voltages. This so-called coupled left shift (CLS) leads to a chronic influx of Na+ into the cell that eventually causes spontaneous or "ectopic" firing along the axon, even in the absence of stimuli. The bifurcations underlying this enhanced excitability have been worked out in full ionic models of this effect. Here, we present computational evidence that increased temperature T can exacerbate this pathological state. Conversely, and perhaps of clinical relevance, mild cooling is shown to move the naturally quiescent cell further away from the threshold of ectopic behavior. The origin of this stabilization-by-cooling effect is analyzed by knocking in and knocking out, one at a time, various processes thought to be T-dependent. The T-dependence of the Na+ current, quantified by its Q 10-Na factor, has the biggest impact on the threshold, followed by Q 10-pump of the sodium-potassium exchanger. Below the ectopic boundary, the steady state for the gating variables and the resting potential are not modified by temperature, since our model separately tallies the Na+ and K+ ions including their separate leaks through the pump. When only the gating kinetics are considered, cooling is detrimental, but in the full T-dependent model, it is beneficial because the other processes dominate. Cooling decreases the pump's activity, and since the pump hyperpolarizes, less hyperpolarization should lead to more excitability and ectopic behavior. But actually the opposite happens in the full model because decreased pump activity leads to smaller gradients of Na+ and K+, which in turn decreases the driving force of the Na+ current.
Assuntos
Axônios , Potenciais da Membrana , Condução Nervosa , Ferimentos e Lesões/fisiopatologia , Animais , Análise por Conglomerados , Humanos , Cinética , Neurônios , Oscilometria , Potássio , Sódio/fisiologia , TemperaturaRESUMO
Spontaneous network activity (SNA) emerges in the spinal cord (SC) before the formation of peripheral sensory inputs and central descending inputs. SNA is characterized by recurrent giant depolarizing potentials (GDPs). Because GDPs in motoneurons (MNs) are mainly evoked by prolonged release of GABA, they likely necessitate sustained firing of interneurons. To address this issue we analyzed, as a model, embryonic Renshaw cell (V1R) activity at the onset of SNA (E12.5) in the embryonic mouse SC (both sexes). V1R are one of the interneurons known to contact MNs, which are generated early in the embryonic SC. Here, we show that V1R already produce GABA in E12.5 embryo, and that V1R make synaptic-like contacts with MNs and have putative extrasynaptic release sites, while paracrine release of GABA occurs at this developmental stage. In addition, we discovered that V1R are spontaneously active during SNA and can already generate several intrinsic activity patterns including repetitive-spiking and sodium-dependent plateau potential that rely on the presence of persistent sodium currents (INap). This is the first demonstration that INap is present in the embryonic SC and that this current can control intrinsic activation properties of newborn interneurons in the SC of mammalian embryos. Finally, we found that 5 µm riluzole, which is known to block INaP, altered SNA by reducing episode duration and increasing inter-episode interval. Because SNA is essential for neuronal maturation, axon pathfinding, and synaptogenesis, the presence of INaP in embryonic SC neurons may play a role in the early development of mammalian locomotor networks.SIGNIFICANCE STATEMENT The developing spinal cord (SC) exhibits spontaneous network activity (SNA) involved in the building of nascent locomotor circuits in the embryo. Many studies suggest that SNA depends on the rhythmic release of GABA, yet intracellular recordings of GABAergic neurons have never been performed at the onset of SNA in the SC. We first discovered that embryonic Renshaw cells (V1R) are GABAergic at E12.5 and spontaneously active during SNA. We uncover a new role for persistent sodium currents (INaP) in driving plateau potential in V1R and in SNA patterning in the embryonic SC. Our study thus sheds light on a role for INaP in the excitability of V1R and the developing SC.
Assuntos
Neurônios GABAérgicos/fisiologia , Rede Nervosa/fisiologia , Células de Renshaw/fisiologia , Canais de Sódio/fisiologia , Sódio/fisiologia , Medula Espinal/embriologia , Potenciais de Ação , Animais , Antagonistas de Aminoácidos Excitatórios/farmacologia , Feminino , Técnicas de Introdução de Genes , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Neurônios Motores/citologia , Comunicação Parácrina , Técnicas de Patch-Clamp , Riluzol/farmacologia , Medula Espinal/citologia , Sinapses/fisiologiaRESUMO
BACKGROUND: Long QT syndrome mutations in the SCN5A gene are associated with an enhanced late sodium current (INa,L) which may lead to pro-arrhythmic action potential prolongation and intracellular calcium dysregulation. We here investigated the dynamic relation between INa,L, intracellular sodium ([Na+]i) and calcium ([Ca2+]i) homeostasis and pro-arrhythmic events in the setting of a SCN5A mutation. METHODS AND RESULTS: Wild-type (WT) and Scn5a1798insD/+ (MUT) mice (age 3-5â¯months) carrying the murine homolog of the SCN5A-1795insD mutation on two distinct genetic backgrounds (FVB/N and 129P2) were studied. [Na+]i, [Ca2+]i and Ca2+ transient amplitude were significantly increased in 129P2-MUT myocytes as compared to WT, but not in FVB/N-MUT. Accordingly, INa,L wassignificantly more enhanced in 129P2-MUT than in FVB/N-MUT myocytes, consistent with a dose-dependent correlation. Quantitative RT-PCR analysis revealed intrinsic differences in mRNA expression levels of the sodium/potassium pump, the sodium/hydrogen exchanger, and sodiumcalcium exchanger between the two mouse strains. The rate of increase in [Na+]i, [Ca2+]i and Ca2+ transient amplitude following the application of the Na+/K+-ATPase inhibitor ouabain was significantly greater in 129P2-MUT than in 129P2-WT myocytes and was normalized by the INa,L inhibitor ranolazine. Furthermore, ranolazine decreased the incidence of pro-arrhythmic calcium after-transients elicited in 129P2-MUT myocytes. CONCLUSIONS: In this study we established a causal link between the magnitude of INa,L, extent of Na+ and Ca2+ dysregulation, and incidence of pro-arrhythmic events in murine Scn5a1798insD/+ myocytes. Furthermore, our findings provide mechanistic insight into the anti-arrhythmic potential of pharmacological inhibition of INa,L in patients with LQT3 syndrome.
Assuntos
Arritmias Cardíacas/genética , Arritmias Cardíacas/fisiopatologia , Cálcio/fisiologia , Líquido Intracelular/fisiologia , Canal de Sódio Disparado por Voltagem NAV1.5/fisiologia , Sódio/fisiologia , Animais , Arritmias Cardíacas/etiologia , Células Cultivadas , Líquido Intracelular/efeitos dos fármacos , Camundongos , Camundongos da Linhagem 129 , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Miócitos Cardíacos/efeitos dos fármacos , Miócitos Cardíacos/fisiologia , Bloqueadores dos Canais de Sódio/farmacologia , Bloqueadores dos Canais de Sódio/uso terapêutico , Trocador de Sódio e Cálcio/efeitos dos fármacos , Trocador de Sódio e Cálcio/fisiologiaRESUMO
BACKGROUND: In fish with external fertilization, two main start-up mechanisms of the path that blocks or activates the spermatozoan motility apparatus are known. The main factor managing the path is osmolality or potassium ion. In burbot from the European and North American population, contradictory findings regarding the factors influencing the onset of spermatozoa motility were reported. The objective of the current study was to determine the effect of potassium and osmolality on the spermatozoa activation of European burbot, Lota lota (Actinopterygii, Gadiformes, Lotidae). Moreover, the influence of pH, as well as sodium ion concentrations on spermatozoa motility was investigated. Seven parameters characterising motility were traced by means of computer-assisted sperm analysis (CASA). PRINCIPAL FINDINGS: The spermatozoa of European burbot are K+ ion-sensitive. A 6-mM KCl solution significantly decreased motility, and above 12-mM (50 mOsm kg-1) totally ceased spermatozoa movement. Sucrose and Na+ solutions inhibited spermatozoa movement only at concentrations > 450-480 mOsm kg-1. Greater differences in the percentage of motile sperm between individuals were noted in solutions containing high concentrations of chemicals triggering sperm motility. The optimum osmolality for spermatozoa motility is in the range of 100-200 mOsm kg-1. The burbot spermatozoa were motile over a wide range of pH values with the best activation at pH 9. CONCLUSION: It was demonstrated that the spermatozoa of European burbot are inhibited by K+ ions similarly as in North American burbot. Other electrolyte and non-electrolyte solutions inhibit spermatozoa movement only if their osmolality is greater than that of the physiological osmolality of seminal plasma. The data provided on basic knowledge of burbot spermatozoa allow to ensure appropriate conditions during artificial reproduction and scientific research.
Assuntos
Gadiformes/fisiologia , Potássio/farmacologia , Potássio/fisiologia , Motilidade dos Espermatozoides/efeitos dos fármacos , Motilidade dos Espermatozoides/fisiologia , Animais , Concentração de Íons de Hidrogênio , Masculino , Concentração Osmolar , Sêmen/fisiologia , Sódio/farmacologia , Sódio/fisiologia , Sacarose/metabolismoRESUMO
Introducción: Es conocido que el sexo es un condicionante de la regulación renal de sodio y de la presión arterial. Material y métodos: Se estudiaron ratas Wistar machos y hembras a los 150 días de vida, con dieta normo o hipersódica (NaCl 1% v.o.) en los últimos cinco. Se determinaron presión arterial media (PAM), natriuresis, filtrado glomerular (VFG), flujo plasmático renal (FPR) y aldosterona plasmática. Se estudió la expresión Na+,K+-ATPasa total (t-NKA) y defosforilada (d-NKA), citocromo P4504A (CYP4A), cotransportadores Na+,K+,2Cl- tipo 2 (NKCC2) y Na+/Cl- (NCC) y por PCR el ARNm de la cadena α1 de NKA (Atp1a1) en corteza y médula renal. Resultados: La PAM fue mayor y la natriuresis menor en los machos bajo ambas dietas. Con ingesta hipersódica la aldosterona bajó en ambos sexos, el VFG fue menor en hembras y el FPR aumentó en machos (4,09 ± 0,17 vs 2,81 ± 0,12 ml/min/gR; p<0,01 vs dieta normosódica). La t-NKA, d-NKA y Atp1a1 en médula fue mayor en machos con ambas dietas. Con ingesta hipersódica, t-NKA en médula y d-NKA en corteza y médula disminuyeron en hembras y solamente d-NKA disminuyó en médula de machos. Asimismo, aumentó CYP4A y disminuyó NKCC2 y NCC en hembras, mientras que aumentó NKCC2, sin cambios en NCC, en machos. Conclusión: El sexo condiciona la presión arterial y el balance de sodio, disminuyendo su reabsorción en hembras y aumentando el FPR en machos. Esto sugiere posibilidades de estudio diferenciales según sexo en trastornos del metabolismo del sodio
Introduction: It is known that sex is a determinant of renal sodium regulation and blood pressure. Methods: Male and female Wistar rats, which were 150 days old and a diet with normal or high levels of sodium (NaCl 1% v.o.), were studied for the last five days. Mean blood pressure (MBP), natriuresis, glomerular filtration rate (GFR), renal plasma flow (RPF) and plasma aldosterone level were established. The following were studied: expressions of total Na+,K+,-ATPase (t-NKA); dephosphorylated NKA (d-NKA); cytochrome P4504A (CYP4A); Na+K+-2Cl- (NKCC2) and Na+/Cl- (NCC) cotransporters. The mRNA expression of the NKA α1 (Atp1a1) chain was examined through PCR analysis in the renal cortex and marrow. Results: Male rats having both types of diet showed higher MBP and lower natriuresis. High sodium intake triggered lower aldosterone levels in both sexes; GFR was lower in females and RPF was higher in males (4.09 ± 0.17 vs. 2.81 ± 0.12 ml/min/gr; p<0.01 vs. diet with a normal sodium level). Marrow t-NKA, d-NKA and Atp1a1 were higher in males on both diets. High sodium intake caused lower marrow t-NKA as well as lower cortex and marrow d-NKA in females. In the case of males, only marrow d-NKA decreased. Furthermore, females showed a higher level of CYP4A and lower levels of NKCC2 and NCC, whereas males showed higher levels of NKCC2 and no variations in NCC. Conclusion: Sex conditions blood pressure and sodium balance, reducing resorption in females and increasing RPF in males. This suggests the possibility of studying sodium metabolism disorders differently according to sex
Assuntos
Animais , Ratos , Sexo , Sódio/fisiologia , Pressão Sanguínea , Ratos WistarRESUMO
The glycosaminoglycan (GAG) molecules are a group of high molecular weight, negatively charged polysaccharides present abundantly in the mammalian organism. By their virtue of ion and water binding capacity, they may affect the redistribution of body fluids and ultimately the blood pressure. Data from the literature suggests that the mitogens Vascular Endothelial Growth Factor (VEGF)-A and VEGF-C are able to regulate the amount and charge density of GAGs and their detachment from the cell surface. Based on these findings we investigated the relationship between the level of dietary sodium intake, the expression levels of VEGF-A and VEGF-C, and the amount of the skin GAGs hyaluronic acid and chondroitin sulfate in an in vivo rat model. Significant correlation between dietary sodium intake, skin sodium levels and GAG content was found. We confirmed the GAG synthesizing role of VEGF-C but failed to prove that GAGs are degraded by VEGF-A. No significant difference in blood pressure was registered between the different dietary groups. A quotient calculated form the ion and water content of the skin tissue samples suggests that - in contrast to previous findings - the osmotically inactive ions and bound water fractions are proportional.
Assuntos
Glicosaminoglicanos/metabolismo , Pele/metabolismo , Sódio na Dieta/administração & dosagem , Sódio/fisiologia , Animais , Feminino , Distribuição Aleatória , Ratos , Ratos Wistar , Pele/efeitos dos fármacos , Fator A de Crescimento do Endotélio Vascular/biossínteseRESUMO
Sodium deficiency increases angiotensin II (ATII) and aldosterone, which synergistically stimulate sodium retention and consumption. Recently, ATII-responsive neurons in the subfornical organ (SFO) and aldosterone-sensitive neurons in the nucleus of the solitary tract (NTSHSD2 neurons) were shown to drive sodium appetite. Here we investigate the basis for NTSHSD2 neuron activation, identify the circuit by which NTSHSD2 neurons drive appetite, and uncover an interaction between the NTSHSD2 circuit and ATII signaling. NTSHSD2 neurons respond to sodium deficiency with spontaneous pacemaker-like activity-the consequence of "cardiac" HCN and Nav1.5 channels. Remarkably, NTSHSD2 neurons are necessary for sodium appetite, and with concurrent ATII signaling their activity is sufficient to produce rapid consumption. Importantly, NTSHSD2 neurons stimulate appetite via projections to the vlBNST, which is also the effector site for ATII-responsive SFO neurons. The interaction between angiotensin signaling and NTSHSD2 neurons provides a neuronal context for the long-standing "synergy hypothesis" of sodium appetite regulation.
Assuntos
Aldosterona/fisiologia , Angiotensina II/fisiologia , Relógios Biológicos/fisiologia , Neurônios/fisiologia , Transdução de Sinais , Sódio/fisiologia , Núcleo Solitário/fisiologia , Animais , Ingestão de Alimentos/fisiologia , Canais Disparados por Nucleotídeos Cíclicos Ativados por Hiperpolarização/fisiologia , Masculino , Camundongos , Camundongos Transgênicos , Canal de Sódio Disparado por Voltagem NAV1.5/fisiologia , Vias Neurais/fisiologia , Núcleos Septais/fisiologia , Sódio/deficiênciaRESUMO
Renal function plays a central role in cardiovascular, kidney, and multiple other diseases, and many existing and novel therapies act through renal mechanisms. Even with decades of accumulated knowledge of renal physiology, pathophysiology, and pharmacology, the dynamics of renal function remain difficult to understand and predict, often resulting in unexpected or counterintuitive therapy responses. Quantitative systems pharmacology modeling of renal function integrates this accumulated knowledge into a quantitative framework, allowing evaluation of competing hypotheses, identification of knowledge gaps, and generation of new experimentally testable hypotheses. Here we present a model of renal physiology and control mechanisms involved in maintaining sodium and water homeostasis. This model represents the core renal physiological processes involved in many research questions in drug development. The model runs in R and the code is made available. In a companion article, we present a case study using the model to explore mechanisms and pharmacology of salt-sensitive hypertension.