Action Potential Characteristics Research Articles

Sex differences in membrane properties and cellular excitability of dopamine D1 receptor-expressing neurons within the shell of the nucleus accumbens of pre- and mid-adolescent mice

BackgroundThe transition from childhood to adulthood, or adolescence, a developmental stage, is characterized by psychosocial and biological changes. The nucleus accumbens (NAc), a striatal brain region composed of the core (NAcC) and shell (NAcSh), has been linked to risk-taking behavior and implicated in reward seeking and evaluation. Most neurons in the NAc are medium spiny neurons (MSNs) that express dopamine D1 receptors (D1R +) and/or dopamine D2 receptors (D2R +). Changes in dopaminergic and glutamatergic systems occur during adolescence and converge in the NAc. While there are previous investigations into sex differences in membrane excitability and synaptic glutamate transmission in both subdivisions of the NAc, to our knowledge, none have specified NAcSh D1R + MSNs from mice during pre- and mid-adolescence.MethodsSagittal brain slices containing the NAc were prepared from B6.Cg-Tg(Drd1a-tdTomato)6Calak/J mice of both sexes from postnatal days 21–25 and 35–47, representing pre- and mid-adolescence, respectively. Whole-cell electrophysiology recordings were collected from NAcSh D1R + MSNs in the form of membrane-voltage responses to current injections, to assess membrane properties and action potential waveform characteristics, and spontaneous excitatory postsynaptic currents (sEPSCs) to assess glutamatergic synaptic activity.ResultsRelative to pre-adolescent males, pre-adolescent female NAcSh D1R + MSNs exhibited a less hyperpolarized resting membrane potential, increased input resistance, and smaller action potential afterhyperpolarization amplitudes. During mid-adolescence, decreased input resistance and a shorter action potential duration in females were the only sex differences observed.ConclusionsTaken together, our results indicate that NAcSh D1R + MSNs in mice exhibit sex differences in membrane properties and AP waveform during pre-adolescence that are overall indicative of increased cellular excitability in females and are suggestive of possible sex differences in glycine receptors, inwardly-rectifying potassium channels, and large conductance voltage-gated potassium channels. These differences do not appear to persist into mid-adolescence, when sex was observed to affect input resistance oppositely to that of pre-adolescence and AP waveform in a manner suggestive of differences in voltage-gated potassium channels.

In vivo calcium and voltage mapping of the zebrafish heart

Abstract Funding Acknowledgements Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Research Foundation Flanders (FWO) Purpose Inherited cardiac arrhythmias are characterized by a disturbance in the normal electrical functioning of the heart due to a dysfunction of cardiac ion channels, accessory proteins and/or structural components. This complex process is difficult to model in vitro, while in vivo mouse models differ substantially from humans with regards to cardiac electrophysiology. Thus, there is a need for alternative disease models in cardiac arrhythmia research. The zebrafish heart shows remarkable similarity to humans in its electrocardiogram and action potential (AP) morphology. Due to the optical translucency of zebrafish larvae, it is possible visualize the cardiac calcium and voltage dynamics across their entire hearts in vivo by use of genetically encoded calcium and voltage indicators (GECI and GEVI). We combined the fast kinetics of a next generation voltage reporter (Ace2N-mNeon) with a spectrally compatible calcium sensor (R-GECO) to create a sensitive high throughput method for in vivo characterization of the cardiac AP. Methods Ace2N-mNeon and R-GECO were expressed under control of a myocardial specific promoter via the Tol2kit system. The effect of sensor expression on normal physiology was assessed by patch clamp experiments. Fluorescence recordings were obtained with a Leica SP8 DLS microscope and a custom-built light sheet microscope. Our zebrafish sensor line was validated by experiments with quinidine drug exposure, as well as a novel model for long QT syndrome (LQTS) with a knock-in (KI) mutation in the cacna1c gene. Results Sensor expression did not significantly alter cardiac electrophysiology. The tg(myl7:Ace2N-mNeon/R-GECO) line demonstrated alterations in the heart rate, depolarization and repolarization induced by quinidine exposure. We observed a significantly prolonged ventricular repolarization in the LQTS KI line. Three-dimensional optical maps of AP characteristics across the entire zebrafish heart exhibited the physiological characteristics of the zebrafish cardiac electrophysiology, with activation initiated at the atrial venous pole and propagated towards the ventricular outflow tract, prolonged repolarization in the ventricles compared to the atria and slowed depolarization in the atrio-ventricular canal. Conclusion By expressing Ace2N-mNeon and R-GECO in the zebrafish heart, we were able to accurately detect drug- and mutation induced alterations in the cardiac AP, as well as AP characteristics across the entire heart, without compromising normal cardiac electrophysiology. Our transgenic line will provide a promising assay for future studies of cardiac arrhythmia.

Change of voltage-gated sodium channel repertoire in skeletal muscle of a MuSK myasthenia gravis mouse model.

Muscle-specific kinase myasthenia gravis (MuSK MG) is caused by autoantibodies against MuSK in the neuromuscular junction (NMJ). MuSK MG patients have fluctuating, fatigable skeletal muscle weakness, in particular of bulbar muscles. Severity differs greatly between patients, in spite of comparable autoantibody levels. One explanation for inter-patient and inter-muscle variability in sensitivity might be variations in compensatory muscle responses. Previously, we developed a passive transfer mouse model for MuSK MG. In preliminary ex vivo experiments, we observed that muscle contraction of some mice, in particular those with milder myasthenia, had become partially insensitive to inhibition by μ-Conotoxin-GIIIB, a blocker of skeletal muscle NaV1.4 voltage-gated sodium channels. We hypothesised that changes in NaV channel expression profile, possibly co-expression of (μ-Conotoxin-GIIIB insensitive) NaV1.5 type channels, might lower the muscle fibre's firing threshold and facilitate neuromuscular synaptic transmission. To test this hypothesis, we here performed passive transfer in immuno-compromised mice, using 'high', 'intermediate' and 'low' dosing regimens of purified MuSK MG patient IgG4. We compared myasthenia levels, μ-Conotoxin-GIIIB resistance and muscle fibre action potential characteristics and firing thresholds. High- and intermediate-dosed mice showed clear, progressive myasthenia, not seen in low-dosed animals. However, diaphragm NMJ electrophysiology demonstrated almost equal myasthenic severities amongst all regimens. Nonetheless, low-dosed mouse diaphragms showed a much higher degree of μ-Conotoxin-GIIIB resistance. This was not explained by upregulation of Scn5a (the NaV1.5 gene), lowered muscle fibre firing thresholds or histologically detectable upregulated NaV1.5 channels. It remains to be established which factors are responsible for the observed μ-Conotoxin-GIIIB insensitivity and whether the NaV repertoire change is compensatory beneficial or a bystander effect.

Decreasing microtubule detyrosination modulates Nav1.5 subcellular distribution and restores sodium current in mdx cardiomyocytes.

The microtubule (MT) network plays a major role in the transport of the cardiac sodium channel Nav1.5 to the membrane, where the latter associates with interacting proteins such as dystrophin. Alterations in MT dynamics are known to impact on ion channel trafficking. Duchenne muscular dystrophy (DMD), caused by dystrophin deficiency, is associated with an increase in MT detyrosination, decreased sodium current (INa), and arrhythmias. Parthenolide (PTL), a compound that decreases MT detyrosination, has shown beneficial effects on cardiac function in DMD. We here investigated its impact on INa and Nav1.5 subcellular distribution. Ventricular cardiomyocytes (CMs) from wild-type (WT) and mdx (DMD) mice were incubated with either 10 µM PTL, 20 µM EpoY, or dimethylsulfoxide (DMSO) for 3-5 h, followed by patch-clamp analysis to assess INa and action potential (AP) characteristics in addition to immunofluorescence and stochastic optical reconstruction microscopy (STORM) to investigate MT detyrosination and Nav1.5 cluster size and density, respectively. In accordance with previous studies, we observed increased MT detyrosination, decreased INa and reduced AP upstroke velocity (Vmax) in mdx CMs compared to WT. PTL decreased MT detyrosination and significantly increased INa magnitude (without affecting INa gating properties) and AP Vmax in mdx CMs, but had no effect in WT CMs. Moreover, STORM analysis showed that in mdx CMs, Nav1.5 clusters were decreased not only in the grooves of the lateral membrane (LM; where dystrophin is localized) but also at the LM crests. PTL restored Nav1.5 clusters at the LM crests (but not at the grooves), indicating a dystrophin-independent trafficking route to this subcellular domain. Interestingly, Nav1.5 cluster density was also reduced at the intercalated disc (ID) region of mdx CMs, which was restored to WT levels by PTL. Treatment of mdx CMs with EpoY, a specific MT detyrosination inhibitor, also increased INa density, while decreasing the amount of detyrosinated MTs, confirming a direct mechanistic link. Attenuating MT detyrosination in mdx CMs restored INa and enhanced Nav1.5 localization at the LM crest and ID. Hence, the reduced whole-cell INa density characteristic of mdx CMs is not only the consequence of the lack of dystrophin within the LM grooves but is also due to reduced Nav1.5 at the LM crest and ID secondary to increased baseline MT detyrosination. Overall, our findings identify MT detyrosination as a potential therapeutic target for modulating INa and subcellular Nav1.5 distribution in pathophysiological conditions.

Optogenetic confirmation of transverse-tubular membrane excitability in intact cardiac myocytes.

T-tubules (TT) form a complex network of sarcolemmal membrane invaginations, essential for well-co-ordinated excitation-contraction coupling (ECC) and thus homogeneous mechanical activation of cardiomyocytes. ECC is initiated by rapid depolarization of the sarcolemmal membrane. Whether TT membrane depolarization is active (local generation of action potentials; AP) or passive (following depolarization of the outer cell surface sarcolemma; SS) has not been experimentally validated in cardiomyocytes. Based on the assessment of ion flux pathways needed for AP generation, we hypothesize that TT are excitable. We therefore explored TT excitability experimentally, using an all-optical approach to stimulate and record trans-membrane potential changes in TT that were structurally disconnected, and hence electrically insulated, from the SS membrane by transient osmotic shock. Our results establish that cardiomyocyte TT can generate AP. These AP show electrical features that differ substantially from those observed in SS, consistent with differences in the density of ion channels and transporters in the two different membrane domains. We propose that TT-generated AP represent a safety mechanism for TT AP propagation and ECC, which may be particularly relevant in pathophysiological settings where morpho-functional changes reduce the electrical connectivity between SS and TT membranes. KEY POINTS: Cardiomyocytes are characterized by a complex network of membrane invaginations (the T-tubular system) that propagate action potentials to the core of the cell, causing uniform excitation-contraction coupling across the cell. In the present study, we investigated whether the T-tubular system is able to generate action potentials autonomously, rather than following depolarization of the outer cell surface sarcolemma. For this purpose, we developed a fully optical platform to probe and manipulate the electrical dynamics of subcellular membrane domains. Our findings demonstrate that T-tubules are intrinsically excitable, revealing distinct characteristics of self-generated T-tubular action potentials. This active electrical capability would protect cells from voltage drops potentially occurring within the T-tubular network.

High-throughput methods for cardiac cellular electrophysiology studies: the road to personalized medicine.

Personalized medicine refers to the tailored application of medical treatment at an individual level, considering the specific genotype or phenotype of each patient for targeted therapy. In the context of cardiovascular diseases, implementing personalized medicine is challenging due to the high costs involved and the slow pace of identifying the pathogenicity of genetic variants, deciphering molecular mechanisms of disease, and testing treatment approaches. Scalable cellular models such as human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) serve as useful in vitro tools that reflect individual patient genetics and retain clinical phenotypes. High-throughput functional assessment of these constructs is necessary to rapidly assess cardiac pathogenicity and test new therapeutics if personalized medicine is to become a reality. High-throughput photometry recordings of single cells coupled with potentiometric probes offer cost-effective alternatives to traditional patch-clamp assessments of cardiomyocyte action potential characteristics. Importantly, automated patch-clamp (APC) is rapidly emerging in the pharmaceutical industry and academia as a powerful method to assess individual membrane-bound ionic currents and ion channel biophysics over multiple cells in parallel. Now amenable to primary cell and hiPSC-CM measurement, APC represents an exciting leap forward in the characterization of a multitude of molecular mechanisms that underlie clinical cardiac phenotypes. This review provides a summary of state-of-the-art high-throughput electrophysiological techniques to assess cardiac electrophysiology and an overview of recent works that successfully integrate these methods into basic science research that could potentially facilitate future implementation of personalized medicine at a clinical level.

Cumulative Spike Train Estimation for Muscle Excitation Assessment From Surface EMG Using Spatial Spike Detection.

Estimating cumulative spike train (CST) of motor units (MUs) from surface electromyography (sEMG) is essential for the effective control of neural interfaces. However, the limited accuracy of existing estimation methods greatly hinders the further development of neural interface. This paper proposes a simple but effective approach for identifying CST based on spatial spike detection from high-density sEMG. Specifically, we use a spatial sliding window to detect spikes according to the spatial propagation characteristics of the motor unit action potential, focusing on the spikes of activated MUs in a local area rather than those of a specific MU. We validated the effectiveness of our proposed method through an experiment involving wrist flexion/extension and pronation/supination, comparing it with a recognized CST estimation method and an MU decomposition based method. The results demonstrated that the proposed method obtained higher accuracy on multi-DoF wrist torque estimation leveraging the estimated CST compared to the other three methods. On average, the correlation coefficient (R) and the normalized root mean square error (nRMSE) between the estimation results and recorded force were 0.96 0.03 and 10.1% 3.7%, respectively. Moreover, there was an extremely high interpretive extent between the CSTs of proposed method and the MU decomposition method. The outcomes reveal the superiority of the proposed method in identifying CSTs and can provide promising driven signals for neural interface.

Neurovirulent cytokines increase neuronal excitability in a model of coronavirus-induced neuroinflammation

BackgroundNeurological manifestations of severe coronavirus infections, including SARS-CoV-2, are wide-ranging and may persist following virus clearance. Detailed understanding of the underlying changes in brain function may facilitate the identification of therapeutic targets. We directly tested how neocortical function is impacted by the specific panel of cytokines that occur in coronavirus brain infection. Using the whole-cell patch-clamp technique, we determined how the five cytokines (TNFα, IL-1β, IL-6, IL-12p40 and IL-15 for 22–28-h) at concentrations matched to those elicited by MHV-A59 coronavirus brain infection, affected neuronal function in cultured primary mouse neocortical neurons.ResultsWe evaluated how acute cytokine exposure affected neuronal excitability (propensity to fire action potentials), membrane properties, and action potential characteristics, as well as sensitivity to changes in extracellular calcium and magnesium (divalent) concentration. Neurovirulent cytokines increased spontaneous excitability and response to low divalent concentration by depolarizing the resting membrane potential and hyperpolarizing the action potential threshold. Evoked excitability was also enhanced by neurovirulent cytokines at physiological divalent concentrations. At low divalent concentrations, the change in evoked excitability was attenuated. One hour after cytokine removal, spontaneous excitability and hyperpolarization of the action potential threshold normalized but membrane depolarization and attenuated divalent-dependent excitability persisted.ConclusionsCoronavirus-associated cytokine exposure increases spontaneous excitability in neocortical neurons, and some of the changes persist after cytokine removal.

The role of acid-sensing ion channels in monosodium urate-induced gouty pain in mice.

Acid-sensing ion channels (ASICs) in dorsal root ganglion (DRG) neurons play an important role in inflammatory pain. The objective of this study is to observe the regulatory role of ASICs in monosodium urate (MSU) crystal-induced gout pain and explore the basis for ASICs in DRG neurons as a target for gout pain treatment. The gout arthritis model was induced by injecting MSU crystals into the ankle joint of mice. The circumference of the ankle joint was used to evaluate the degree of swelling; the von Frey filaments were used to determine the withdrawal threshold of the paw. ASIC currents and action potentials (APs) were recorded by patch clamp technique in DRG neurons. The results displayed that injecting MSU crystals caused ankle edema and mechanical hyperalgesia of the paw, which was relieved after amiloride treatment. The ASIC currents in DRG neurons were increased to a peak on the second day after injecting MSU crystals, which were decreased after amiloride treatment. MSU treatment increased the current density of ASICs in different diameter DRG cells. MSU treatment does not change the characteristics of AP. The results suggest that ASICs in DRG neurons participate in MSU crystal-induced gout pain.

Sustained high-fat diet increases ventricular susceptibility to arrhythmias and impairs myocardial calcium cycling

Abstract Funding Acknowledgements Type of funding sources: Public Institution(s). Main funding source(s): Medizinische Fakultät, Heinrich Heine Universität Düsseldorf Background – Obesity is one of the most relevant factors for cardiovascular disease in western societies. Independent of coronary heart disease, obesity promotes a multimodal arrhythmic substrate and increases the susceptibility to ventricular arrhythmias (VA) eventually leading to sudden cardiac death. Purpose – Aim of the present study was to develop an ex vivo mouse model to characterize obesity-mediated VAs and left ventricular (LV) myocardial disruption of calcium homeostasis. Methods – All experiments were performed ex vivo in retrogradely perfused hearts from 32 – 36 week-old C57Bl/6J mice (n=14). Five mice received a normal chow diet (NCD), 9 mice were fed a sustained high-fat diet (HFD) over a 6-month period to generate diet-induced-obesity (DIO). We combined an endocardial catheter-based technique to assess VA susceptibility with an epicardial dual fluorescence imaging approach employing a potentiometric (Rh237) and a calcium sensitive (Rhod2) dye to assess action potential and calcium cycling characteristics. Optical mapping data was analysed using a MATLAB-based algorithm. VA susceptibility was quantified by an established scoring system. Results – Mice receiving a sustained HFD were significantly heavier than those receiving NCD (NCD: 34.8±2.1g, HFD: 53.3±3.2g, P<0.0001). The number of induced VAs and VA susceptibility were increased in HFD compared to NCD (number of induced VAs: NCD: 1.4±0.7, HFD: 6.3±0.8, P=0.005, Figure 1A; VA susceptibility: NCD: 1.4±0.7, HFD: 18.1±2.3, P=0.001, Figure 1B). Ventricular tachycardias (VT) were only inducible in HFD, additionally, those hearts presented mostly polymorphic VTs (VT inducibility: NCD: 0.0 %, HFD: 88.9 %, P=0.003, Figure 1C; VT morphology: monomorphic VT: 25 %, polymorphic VT: 75 %). Epicardial optical mapping of the anterior LV wall showed a prolonged action potential duration (APD) and prolonged calcium transient duration (CaTD) in HFD (APD90 at CL 100 ms: NCD: 45.1±1.5 ms, HFD: 50.8±1.8 ms, P=0.03; CaTD90 at 100 ms pacing cycle length (PCL): NCD: 61.0±2.3 ms, HFD: 66.6±0.6 ms, P=0.02, Figure 2A). Furthermore, CaT alternans (ALT) threshold defined as amplitude alternans > 10 % of two consecutive beats was decreased (PCL ALT threshold: NCD: 60.0±1.6 ms, DIO: 67.8±1.2 ms, P=0.006, Figure 2B), with mean ALT being increased in HFD (mean ATL at 60 ms PCL: NCD: 15.4±2.4 %, HFD: 25.7±3.0 %, P=0.04). Conclusions – Our findings show that sustained HFD in mice increases ventricular susceptibility to VTs, especially polymorphic VTs indicating heterogeneity in wave propagation. DIO leads to disruption of LV myocardial calcium homeostasis as demonstrated by a decreased ALT threshold and increased mean ALT. Our data suggest that LV calcium mishandling may be a driver for arrhythmogenesis in DIO and modulation may present a perspective therapeutical approach. Further experiments will address molecular mechanisms and identify potential targets of modulation.

Decreasing microtubule detyrosination by parthenolide restores sodium current in mdx cardiomyocytes

Abstract Funding Acknowledgements Type of funding sources: None. Background The cardiac sodium channel Nav1.5 is transported to the membrane by the microtubule network. Alterations in microtubule dynamics are known to impact on ion channel trafficking. Pathophysiological conditions such as heart failure and Duchenne muscular dystrophy (DMD) are associated with an increase in microtubule detyrosination as well as a decreased sodium current (INa) and pro-arrhythmia. Parthenolide, a compound that decreases the fraction of detyrosinated microtubules, has been shown to have beneficial effects on cardiac function in DMD mice, but its impact on INa has not been investigated. Methods and Results Cardiomyocytes (CMs) from wild type (WT) and mdx (DMD) mice were used to investigate the effect of parthenolide. Cells were incubated with either 10 µM parthenolide or DMSO for 3-5 hours. INa and action potential (AP) characteristics were assessed using the patch-clamp technique, while immunofluorescence and stochastic optical reconstruction microscopy (STORM), were used to investigate microtubule detyrosination and Nav1.5 cluster density, respectively. In accordance with previous studies, we observed increased levels of detyrosinated microtubules and decreased INa in mdx CMs compared to WT. Treatment with parthenolide decreased the fraction of detyrosinated microtubules and significantly increased INa magnitude in mdx CMs, but had no effect on INa in WT CMs. Accordingly, parthenolide significantly increased AP maximal upstroke velocity in mdx CMs, without affecting other AP properties. Parthenolide did not affect INa gating properties, indicating that it increased INa by enhancing Nav1.5 membrane trafficking. Indeed, STORM analysis showed that parthenolide increased Nav1.5 cluster density at both the lateral membrane (crest) and intercalated disc region in mdx CMs while it had no effect on WT CMs. Conclusions Parthenolide restores INa and Nav1.5 membrane expression in mdx CMs, and may be of potential benefit in other pathophysiological conditions associated with increased microtubule detyrosination and reduced INa. Further elucidation of the impact of microtubule dynamics on Nav1.5 may identify additional therapeutic targets for restoring conduction and preventing arrhythmias.

Cardiac deficiency of P21-activated kinase 1 promotes atrial arrhythmogenesis in mice following adrenergic challenge.

P21-activated kinase 1 (Pak1) signalling plays a vital and overall protective role in the heart. However, the phenotypes of Pak1 deficiency in the cardiac atria have not been well explored. In this study, Pak1 cardiac-conditional knock-out (cKO) mice were studied under baseline and adrenergic challenge conditions. Pak1 cKO mice show atrial arrhythmias including atrial fibrillation (AF) in vivo, detected during anaesthetized electrocardiography without evidence of interstitial fibrosis upon Masson's trichrome staining. Optical mapping of left atrial preparations from Pak1 cKO mice revealed a higher incidence of Ca2+ and action potential alternans under isoprenaline challenge and differences in baseline action potential and calcium transient characteristics. Type-2 ryanodine receptor (RyR2) channels from Pak1 cKO hearts had a higher open probability than those from wild-type. Reverse transcription-quantitative polymerase chain reaction and Western blotting indicated that pCamkIIδ and RyR2 are highly phosphorylated at baseline in the atria of Pak1 cKO mice, while the expression of Slc8a2 and Slc8a3 as a Na+-Ca2+ exchanger, controlling the influx of Ca2+ from outside of the cell and efflux of Na+ from the cytoplasm, are augmented. Chromatin immunoprecipitation study showed that pCreb1 interacts with Slc8a2 and Slc8a3. Our study thus demonstrates that deficiency of Pak1 promotes atrial arrhythmogenesis under adrenergic stress, probably through post-translational and transcriptional modifications of key molecules that are critical to Ca2+ homeostasis. This article is part of the theme issue 'The heartbeat: its molecular basis and physiological mechanisms'.

Differential Sodium Current Remodelling Identifies Distinct Cellular Proarrhythmic Mechanisms in Paroxysmal vs Persistent Atrial Fibrillation.

The cellular mechanisms underlying progression from paroxysmal to persistent atrial fibrillation (AF) are not fully understood, but alterations in (late) sodium current (INa) have been proposed. Human studies investigating electrophysiological changes at the paroxysmal stage of AF are sparse, with the majority employing right atrial appendage cardiomyocytes (CMs). We here investigated action potential (AP) characteristics and (late) INa remodelling in left atrial appendage CMs (LAA-CMs) from patients with paroxysmal and persistent AF and patients in sinus rhythm (SR), as well as the potential contribution of the "neuronal" sodium channel SCN10A/NaV1.8. Peak INa, late INa and AP properties were investigated through patch-clamp analysis on single LAA-CMs, whereas quantitative polymerase chain reaction was used to assess SCN5A/SCN10A expression levels in LAA tissue. In paroxysmal and persistent AF LAA-CMs, AP duration was shorter than in SR LAA-CMs. Compared with SR, peak INa and SCN5A expression were significantly decreased in paroxysmal AF, whereas they were restored to SR levels in persistent AF. Conversely, although late INa was unchanged in paroxysmal AF compared with SR, it was significantly increased in persistent AF. Peak or late Nav1.8-based INa was not detected in persistent AF LAA-CMs. Similarly, expression of SCN10A was not observed in LAAs at any stage. Our findings demonstrate differences in (late) INa remodeling in LAA-CMs from patients with paroxysmal vs persistent AF, indicating distinct cellular proarrhythmic mechanisms in different AF forms. These observations are of particular relevance when considering potential pharmacologic approaches targeting (late) INa in AF.

A VO2 Neuristor Based on Microstrip Line Coupling.

The neuromorphic network based on artificial neurons and synapses can solve computational difficulties, and its energy efficiency is incomparable to the traditional von Neumann architecture. As a new type of circuit component, nonvolatile memristors are very similar to biological synapses in structure and function. Only one memristor can simulate the function of a synapse. Therefore, memristors provide a new way to build hardware-based artificial neural networks. To build such an artificial neural network, in addition to the artificial synapses, artificial neurons are also needed to realize the distribution of information and the adjustment of synaptic weights. As the VO2 volatile local active memristor is complementary to nonvolatile memristors, it can be used to simulate the function of neurons. However, determining how to better realize the function of neurons with simple circuits is one of the current key problems to be solved in this field. This paper considers the influence of distribution parameters on circuit performance under the action of high-frequency and high-speed signals. Two Mott VO2 memristor units are connected and coupled with microstrip lines to simulate the Hodgkin-Huxley neuron model. It is found that the proposed memristor neuron based on microstrip lines shows the characteristics of neuron action potential: amplification and threshold.

A Novel and Efficient Surface Electromyography Decomposition Algorithm Using Local Spatial Information.

Motor unit spike trains (MUSTs) decomposed from surface electromyography (sEMG) have been an emerging solution for neural interfacing, especially for the control of upper limb prosthetics. Accurate and efficient decomposition techniques are essential and desirable. However, most decomposition methods are designed for motor units (MUs) with global maximum of single or large muscle, while in general forearm muscles are usually small and slender with low global energy. Thus, we propose a novel approach using local spatial information towards more accurate and efficient sEMG decomposition of forearm muscles. A fast spatial spike detection method is proposed to replace the time-consuming iteration process of blind source separation (BSS) methods. Here, spatial distribution characteristics of motor unit action potential are leveraged to pre-classify the candidate MUs, and further to create initial MU templates, aiming to avoid repeating convergence to high-energy MUs. The results of both simulated and experimental sEMG signals show that low-energy MUs from small muscles are more easily found compared with conventional BSS algorithm. Specifically, the proposed method can identify more 40% reliable MUs while only 30% consuming time are needed. The outcomes provide a novel solution for more efficient sEMG decomposition, potentially paving the way of MUST-based non-invasive neural interface.

Comparison of electrophysiological properties of parvalbumin neurons in the tail of the striatum and the auditory cortex of mice

To study the electrophysiological properties of parvalbumin (PV) neurons in the auditory cortex (AC) and its descending auditory projection area in the tail of the striatum (TS). The stimulation response of PV neuron step current was recorded in PV-Cre-Ai14 mice using in vitro patch clamp technique, and the release characteristics and waveform characteristics of PV neuron action potentials (APs) were analyzed using Clampfit and MATLAB software. The release characteristics of the APs included AP onset, rheobase, average firing rate, F/I slope and spike frequency adaptation (SFA); the waveform characteristics included peak and post potential characteristics. The PV neurons of the TS and the AC had significantly different electrophysiological characteristics. In terms of peak potential characteristics, the PV neurons in the TS presented with smaller half peak width (P < 0.001) and larger amplitude (P < 0.01) with larger maximum ascending slope (P < 0.01) and maximum descending slope (P < 0.05). For post potential characteristics, the PV neurons in the TS showed a greater post hyperpolarization (P < 0.01) with a shorter time for recovery of the resting potential (P < 0.01). The firing characteristics of the PV neurons of the TS featured a higher AP rheobase (P < 0.01), a larger F/I slope (P < 0.01), a greater firing onset delay (P < 0.001), and a larger SFA (P < 0.01). The PV neurons in the TS and the AC of mice show significantly different electrophysiological characteristics in processing auditory information.

Extracellular local field potentials in men and women based on a theoretical model of voltage decrease as a function of distance

Extracellular microelectrodes determine the field potential difference adjacent to the neuron and depend on the composition of the tissue, including ohmic resistance and distance from the source. Based on theoretical aspects, a reciprocal voltage versus distance model is postulated, which was assessed by means of clinical neurographical data in men and women. Data of women showed higher voltages and could be slightly better fitted to the model. Voltage amplitude and to a lesser extent rise time (RT) were the only action potential characteristics which could predict the gender in a discriminant analysis model. In conclusion, neurographical studies may be a suitable option to assess ohmic resistance of tissue and composition of tissue in addition to bioimpedance (BIA) methods.

Genetic inhibition of serum glucocorticoid kinase 1 prevents obesity-related atrial fibrillation.

Obesity is an important risk factor for atrial fibrillation (AF), but a better mechanistic understanding of obesity-related atrial fibrillation is required. Serum glucocorticoid kinase 1 (SGK1) is a kinase positioned within multiple obesity-related pathways, and prior work has shown a pathologic role of SGK1 signaling in ventricular arrhythmias. We validated a mouse model of obesity-related AF using wild-type mice fed a high-fat diet. RNA sequencing of atrial tissue demonstrated substantial differences in gene expression, with enrichment of multiple SGK1-related pathways, and we showed upregulated of SGK1 transcription, activation, and signaling in obese atria. Mice expressing a cardiac specific dominant-negative SGK1 were protected from obesity-related AF, through effects on atrial electrophysiology, action potential characteristics, structural remodeling, inflammation, and sodium current. Overall, this study demonstrates the promise of targeting SGK1 in a mouse model of obesity-related AF.

Empagliflozin and dapagliflozin increase Na current in human cardiomyocytes

Abstract Background Empagliflozin (EMPA) and dapagliflozin (DAPA) are sodium-glucose cotransporter 2 inhibitors (SGLT2i) used for the treatment of type 2 Diabetes Mellitus (T2DM). Both drugs reduce morbidity and mortality in heart failure (HF) patients with reduced or preserved ejection fraction, even in the absence of T2DM. Moreover, these drugs decrease ventricular arrhythmias and sudden cardiac death in HF patients. The sodium current (INa), carried by Nav1.5 channels, is responsible for cardiac action potential (AP) depolarization and determines excitability and conduction velocity. In HF patients, the expression of Nav1.5 channels is reduced, leading to a decrease of ventricular excitability that enhances the arrhythmic risk. Purpose We aimed to determine the effects of EMPA and DAPA on human cardiac INa and AP characteristics. Methods Peak INa and ventricular-like APs were recorded in cardiomyocytes derived from human induced pluripotent stem cells (hiPSC-CM) using patch-clamp techniques. INa was also recorded in CHO cells transiently transfected with human Nav1.5+Navβ1 channels. In all cases, EMPA or DAPA (1 μM) were added to culture media and incubated for 24-h. Results APs recorded in hiPSC-CMs exhibited automatic activity and incubation with EMPA or DAPA did not modify spontaneous beating frequency (0.39±0.04 Hz; P&amp;gt;0.05, n≥16). In cells driven at 1 Hz, none of the drugs modified resting membrane potential (−76.7±1.4 mV; P&amp;gt;0.05, n≥11), but significantly increased AP amplitude from 98.6±3.6 to 105±2.2 (DAPA) and 107±2.3 mV (EMPA) (P&amp;lt;0.05). Interestingly, only EMPA lengthened AP duration measured at 20%, 50%, and 90% (from 605.6±31.3 to 760.5±59.0 ms, P&amp;lt;0.05) of repolarization. In hiPSC-CMs EMPA increased maximum INa density by 64% (from −156.0±28.0 to −256.4±28.1 pA/pF, P&amp;lt;0.05, n≥7) and shifted the midpoint (Vh) of the inactivation curve to more hyperpolarized potentials (from −97.3±4.5 to −108.6±4.4 mV, P&amp;lt;0.05, n≥7). In turn, DAPA increased maximum INa density by 24% (to −193.8±26.6 pA/pF) and shifted the Vh of the activation curve to more negative potentials (from −47.2±1.6 mV to −55.5±2.8 mV, P&amp;lt;0.05), an effect that would increase the INa at negative potentials coinciding with channel opening. None of the drugs modified the time course of current activation or inactivation. In CHO cells, EMPA and DAPA effects on INa were identical to those observed on hiPSC-CM. These results suggest that both SGLT2i increase INa by enhancing Nav1.5 expression into the cell membrane, by a direct gating effect on the channel, or by a combination of both. Conclusions In human cardiomyocytes, EMPA and DAPA increase INa and the AP amplitude. Moreover, EMPA, but not DAPA, prolonged AP duration. We propose that EMPA and DAPA exhibit a unique mechanism that increases cardiac excitability and conduction velocity and could contribute to the prevention of arrhythmic events in HF patients. Funding Acknowledgement Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Ministerio de Ciencia e innovaciόnInstituto de de Salud Carlos III

QTc interval and ventricular action potential prolongation in the Mecp2Null/+ murine model of Rett syndrome.

Rett Syndrome (RTT) is a congenital, X‐chromosome‐linked developmental disorder characterized by developmental delay, dysautonomia, and breathing irregularities. RTT is also associated with sudden death and QT intervals are prolonged in some RTT patients. Most individuals with RTT have mutations in the MECP2 gene. Whilst there is some evidence for QT prolongation in mouse models of RTT, there is comparatively little information on how loss of Mecp2 function affects ventricular action potentials (APs) and, to‐date, none on ventricular APs from female RTT mice. Accordingly, the present study was conducted to determine ECG and ventricular AP characteristics of Mecp2 Null/+ female mice. ECG recordings from 12–13 month old female Mecp2 Null/+ mice showed prolonged rate corrected QT (QTc) intervals compared to wild‐type (WT) controls. Although Mecp2 Null/+ animals exhibited longer periods of apnoea than did controls, no correlation between apnoea length and QTc interval was observed. Action potentials (APs) from Mecp2 Null/+ myocytes had longer APD90 values than those from WT myocytes and showed augmented triangulation. Application of the investigational INa,Late inhibitor GS‐6615 (eleclazine; 10 μM) reduced both APD90 and AP triangulation in Mecp2 Null/+ and WT myocytes. These results constitute the first direct demonstration of delayed repolarization in Mecp2 Null/+ myocytes and provide further evidence that GS‐6615 may have potential as an intervention against QT prolongation in RTT.