Linear Current-voltage Relationship Research Articles

Insight into deposition duration-varied crystallinity, optical parameters, morphology and electrical properties of solution-processed nanostructured CdSSe thin films

Nanostructured cadmium sulpho selenide (CdSSe) films have been grown onto glass substrates for different deposition durations via a facile solution-based method known as chemical bath deposition (CBD). XRD studies revealed that the crystallites in the films possessed hexagonal (wurtzite) structures with predominant orientation along the (002) plane. The crystallite size increased from ∼5.95 nm to ∼16.69 nm with the increase in deposition duration from 1 h to 4 h. The absorbance, transmittance and reflectance measurements were carried out. A transparency of ∼(50–80) % was obtained for the films. The direct optical energy band gap was found to decrease from 1.982 eV to 1.952 eV with the increment of deposition duration from 1 h to 4 h. SEM micrographs of the films revealed the presence of nano spherical-shaped grains with an average size ranging from ∼305 nm to ∼612 nm. HRTEM images illustrated the presence of compact nano-spherical grains and nano-crystallites distributed in random orientations. Raman spectroscopy divulged the presence of CdSe and CdS vibrational Raman modes and overtones in the films. PL spectra revealed a prominent red emission band along with weak luminescent bands in the yellow-red emission region. The presence of Cd–Se and Cd–S bonds in the films was validated using FTIR analysis. Electrical investigation revealed the linear current‒voltage relationship for the films with the rise in conductivity with deposition duration. Hence, deposition duration stands out as a vital parameter for modulating the properties of CdSSe thin films for use in photovoltaic devices and optoelectronics.

Non-trivial dynamics in a model of glial membrane voltage driven by open potassium pores

Despite the molecular evidence that a nearly linear steady-state current-voltage relationship in mammalian astrocytes reflects a total current resulting from more than one differentially regulated K+ conductance, detailed ordinary differential equation (ODE) models of membrane voltage Vm are still lacking. Various experimental results reporting altered rectification of the major Kir currents in glia, dominated by Kir4.1, have motivated us to develop a detailed model of Vm dynamics incorporating the weaker potassium K2P-TREK1 current in addition to Kir4.1, and study the stability of the resting state Vr. The main question is whether, with the loss of monotonicity in glial I-V curve resulting from altered Kir rectification, the nominal resting state Vr remains stable, and the cell retains the trivial, potassium electrode behavior with Vm after EK. The minimal two-dimensional model of Vm near Vr showed that an N-shape deformed Kir I-V curve induces multistability of Vm in a model that incorporates K2P activation kinetics, and nonspecific K+ leak currents. More specifically, an asymmetrical, nonlinear decrease of outward Kir4.1 conductance, turning the channels into inward rectifiers, introduces instability of Vr. That happens through a robust bifurcation giving birth to a second, more depolarized stable resting state Vdr > −10 mV. Realistic recordings from electrographic seizures were used to perturb the model. Simulations of the model perturbed by constant current through gap junctions and seizure-like discharges as local field potentials led to depolarization and switching of Vm between the two stable states, in a downstate-upstate manner. In the event of prolonged depolarizations near Vdr, such catastrophic instability would affect all aspects of the glial function, from metabolic support to membrane transport, and practically all neuromodulatory roles assigned to glia.

The oxidation enhancement photocurrent response in WSe1.95Te0.05 nanosheets

The photocurrent was detected in the WSe1.95Te0.05 with and without surface oxidation. The linear current–voltage relation reveal the WSe1.95Te0.05 nanosheet/Pt ohmic contact. The XPS spectrum reveals the WO3 was formed during the oxidation. The photo responsivity increase from 173 A/W to 442 A/W at 0.5 V after air exposuring in ambient conditions over half year. This value is the highest observed values in two-dimensional transition metal dichalcogenides without artificial treatments. The greatly photocurrent enhancement originated from the incident photo-to-current efficiency (IPCE) enhancement of WO3 at the wavelength of 405 nm.

P-Type Ohmic Contact to Monolayer WSe2 Field-Effect Transistors Using High-Electron Affinity Amorphous MoO3

Monolayer tungsten diselenide (1L-WSe2) has been widely used for studying emergent physics due to the unique properties of its valence bands. However, electrical transport studies have been impeded by the lack of a reliable method to realize Ohmic hole-conducting contacts to 1L-WSe2 especially at low carrier densities and low temperatures. Here, we report low-temperature p-type Ohmic contact to 1L-WSe2 field-effect transistors at carrier densities (n) below n = 1 × 1012 cm–2 with negligible temperature dependence down to the lowest measured temperature (10 K). The non-rectifying barrier is achieved between 1L-WSe2 and molybdenum trioxide (MoO3), where 1L-WSe2 underneath MoO3 is heavily hole-doped through surface transfer doping. Electrical transport measurements reveal linear current–voltage relations at a temperature of 10 K and carrier densities from n = 7.7 × 1011 cm–2 to below the threshold. The finding is also supported by nearly temperature-independent output curves up to room temperature and a negligible contact barrier down to the subthreshold regime. Furthermore, the contact resistivity of MoO3-contacted 1L-WSe2 FET is 30.2–64.8 kΩ μm at n = 1.5 × 1012 cm–2, which is the lowest reported for 1L-WSe2 FETs at such low carrier density. Realizing robust p-type Ohmic contact to a 2D transition metal dichalcogenide semiconductor will enable direct electronic measurements of quantum transport in correlated phases in the valence bands of monolayer semiconductors.

Timing Selector: Using Transient Switching Dynamics to Solve the Sneak Path Issue of Crossbar Arrays

Sneak path current is a fundamental issue and a major roadblock to the wide application of memristor crossbar arrays. Traditional selectors such as transistors compromise the 2D scalability and 3D stack‐ability of the array, while emerging selectors with highly nonlinear current–voltage relations contradict the requirement of a linear current–voltage relation for efficient multiplication by directly using Ohm's law. Herein, the concept of a timing selector is proposed and demonstrated, which addresses the sneak path issue with a voltage‐dependent delay time of its transient switching behavior, while preserving a linear current–voltage relationship for computation. Crossbar arrays with silver‐based diffusive memristors as the timing selectors are built and the operation principle and operational windows are experimentally demonstrated. The timing selector enables large memristor crossbar arrays that can be used to solve large‐dimension real‐world problems in machine intelligence and neuromorphic computing.

Development of Highly Sensitive Strain Sensor Using Area-Arrayed Graphene Nanoribbons.

In this study, a basic design of area-arrayed graphene nanoribbon (GNR) strain sensors was proposed to realize the next generation of strain sensors. To fabricate the area-arrayed GNRs, a top-down approach was employed, in which GNRs were cut out from a large graphene sheet using an electron beam lithography technique. GNRs with widths of 400 nm, 300 nm, 200 nm, and 50 nm were fabricated, and their current-voltage characteristics were evaluated. The current values of GNRs with widths of 200 nm and above increased linearly with increasing applied voltage, indicating that these GNRs were metallic conductors and a good ohmic junction was formed between graphene and the electrode. There were two types of GNRs with a width of 50 nm, one with a linear current–voltage relationship and the other with a nonlinear one. We evaluated the strain sensitivity of the 50 nm GNR exhibiting metallic conduction by applying a four-point bending test, and found that the gauge factor of this GNR was about 50. Thus, GNRs with a width of about 50 nm can be used to realize a highly sensitive strain sensor.

Upregulation of transient receptor potential melastatin 4 (TRPM4) in ventricular fibroblasts from heart failure patients.

The transient receptor potential melastatin 4 (TRPM4) is a Ca2+-activated nonselective monovalent cation channel belonging to the TRP channel superfamily. TRPM4 is widely expressed in various tissues and most abundantly expressed in the heart. TRPM4 plays a critical role in cardiac conduction. Patients carrying a gain-of-function or loss-of-function mutation of TRPM4 display impaired cardiac conduction. Knockout or over-expression of TRPM4 in mice recapitulates conduction defects in patients. Moreover, recent studies have indicated that TRPM4 plays a role in hypertrophy and heart failure. Whereas the role of TRPM4 mediated by cardiac myocytes has been well investigated, little is known about TRPM4 and its role in cardiac fibroblasts. Here we show that in human left ventricular fibroblasts, TRPM4 exhibits typical Ca2+-activation characteristics, linear current-voltage (I-V) relation, and monovalent permeability. TRPM4 currents recorded in fibroblasts from heart failure patients (HF) are more than 2-fold bigger than those from control individuals (CTL). The enhanced functional TRPM4 in HF is not resulted from changed channel properties, as TRPM4 currents from both HF and CTL fibroblasts demonstrate similar sensitivity to intracellular calcium activation and extracellular 9-phenanthrol (9-phen) blockade. Consistent with enhanced TRPM4 activity, the protein level of TRPM4 is about 2-fold higher in HF than that of CTL hearts. Moreover, TRPM4 current in CTL fibroblasts is increased after 24 hours of TGFβ1 treatment, implying that TRPM4 in vivo may be upregulated by fibrogenesis promotor TGFβ1. The upregulated TRPM4 in HF fibroblasts suggests that TRPM4 may play a role in cardiac fibrogenesis under various pathological conditions.

Spadin Modulates Astrocytic Passive Conductance via Inhibition of TWIK-1/TREK-1 Heterodimeric Channels.

Astrocytes, the most abundant cell type in the brain, are non-excitable cells and play critical roles in brain function. Mature astrocytes typically exhibit a linear current–voltage relationship termed passive conductance, which is believed to enable astrocytes to maintain potassium homeostasis in the brain. We previously demonstrated that TWIK-1/TREK-1 heterodimeric channels mainly contribute to astrocytic passive conductance. However, the molecular identity of astrocytic passive conductance is still controversial and needs to be elucidated. Here, we report that spadin, an inhibitor of TREK-1, can dramatically reduce astrocytic passive conductance in brain slices. A series of gene silencing experiments demonstrated that spadin-sensitive currents are mediated by TWIK-1/TREK-1 heterodimeric channels in cultured astrocytes and hippocampal astrocytes from brain slices. Our study clearly showed that TWIK-1/TREK-1-heterodimeric channels can act as the main molecular machinery of astrocytic passive conductance, and suggested that spadin can be used as a specific inhibitor to control astrocytic passive conductance.

Examination of Nonlinear Conductivity on the Excitonic Insulator Candidate Ta2NiSe5

The excitonic insulator candidate Ta2NiSe5 has been examined by means of the response to a pulse electric field. We observed linear current–voltage relationship at 200 V/cm, indicating the robustne...

TWIK-1 channels contribute to hypokalemia induced paradoxical depolarization in cardiac myocytes

TWIK-1 potassium channels contributes to background K+ conductance in diverse cell types including human cardiac myocytes. TWIK-1 potassium channels changes ion selectivity and conduct inward Na+ currents at sub-physiological [K+]e. However, the pathological implications of the functional changes of TWIK-1 channels remains largely unknown. In hypokalemia, human cardiomyocytes show paradoxical depolarization which cause cardiac arrhythmia. However, the underlying mechanism is unknown. In the present study, we show that TWIK-1 channels conduct inward leak Na+ currents and contribute to the paradoxical membrane potential depolarization in cardiac myocytes in hypokalemia. First, under hypokalemia conditions, Chinese hamsterovary cells that heterologously express TWIK-1-WT channels show depolarization; in contrast, the cells with expression of TWIK-1-T118I mutant channels show hyperpolarization. Second, both human cardiomyocytes derived from induced pluripotent stem cells with enhanced Kir2.1 expression and mouse HL-1 cardiomyocytes with ectopic expression of TWIK-1 channels show paradoxical depolarization phenomenon. These cardiomyocytes display a nearly linear current-voltage relationship with a reversal potential at about -30 mV at 1 mM [K+]e, indicating a cation conductance. Substitution extra-cellular Na+ with NMDG+ shift the reversal potential to -110 mV and abolish the paradoxical depolarization phenomenon. Besides, inhibition of K2P1 expression eliminates the phenomenon, suggesting contributions of TWIK-1 channels to paradoxical depolarization. These findings reveal the mechanisms underlying hypokalemia induced paradoxical depolarization in human cardiac myocytes, and support that the inward leak Na+ currents carried by TWIK-1 channels induce paradoxical depolarization in cardiac myocytes under hypokalemia conditions.

Ohmic contact formation mechanisms of TiN film on 4H–SiC

The atomic structure, interfacial charge distribution, bonding nature, and interfacial electronic states of a 4H–SiC/TiN interface are systematically investigated to understand the Ohmic contact formation mechanisms of TiN to 4H–SiC. The experiment results clearly demonstrate that the well-arranged TiN (111)-oriented lattice planes are parallel to the (0001) SiC-oriented substrate, which is in line with the XRD results. In addition, the interface is coherent without any secondary phase layers, amorphous layers, or transition regions, which confirms the direct contact of TiN to SiC at the atomic scale, exhibiting a linear current–voltage relationship. Quantitatively, first-principle calculations reveal that the Schottky barrier height (SBH) is as low as 0.03 eV and that the band gap nearly vanishes at the interface, indicating an excellent Ohmic contact of TiN to 4H–SiC. Furthermore, the SBH is significantly reduced through the interfacial charge polarization effect and strong coupling of interfacial electronic states, enhancing the quantum electron transport. The present results provide insight into the complicated electronic effects of the Ohmic contact interface and indicate that TiN is a promising SiC Ohmic contact material for advanced next-generation power device applications.

Functional expression of CLIFAHDD and IHPRF pathogenic variants of the NALCN channel in neuronal cells reveals both gain- and loss-of-function properties

The excitability of neurons is tightly dependent on their ion channel repertoire. Among these channels, the leak sodium channel NALCN plays a crucial role in the maintenance of the resting membrane potential. Importantly, NALCN mutations lead to complex neurodevelopmental syndromes, including infantile hypotonia with psychomotor retardation and characteristic facies (IHPRF) and congenital contractures of limbs and face, hypotonia and developmental delay (CLIFAHDD), which are recessively and dominantly inherited, respectively. Unfortunately, the biophysical properties of NALCN are still largely unknown to date, as well as the functional consequences of both IHPRF and CLIFAHDD mutations on NALCN current. Here we have set-up the heterologous expression of NALCN in the neuronal cell line NG108-15 to investigate the electrophysiological properties of NALCN carrying representative IHPRF and CLIFAHDD mutations. Several original properties of the wild-type (wt) NALCN current were retrieved: mainly carried by external Na+, blocked by Gd3+, insensitive to TTX and potentiated by low external Ca2+ concentration. However, we found that this current displays a time-dependent inactivation in the −80/−40 mV range of membrane potential, and a non linear current-voltage relationship indicative of voltage sensitivity. Importantly, no detectable current was recorded with the IHPRF missense mutation p.Trp1287Leu (W1287L), while the CLIFAHDD mutants, p.Leu509Ser (L509S) and p.Tyr578Ser (Y578S), showed higher current densities and slower inactivation, compared to wt NALCN current. This study reveals that heterologous expression of NALCN channel can be achieved in the neuronal cell line NG108-15 to study the electrophysiological properties of wt and mutants. From our results, we conclude that IHPRF and CLIFAHDD missense mutations are loss- and gain-of-function variants, respectively.

A Compact Thévenin Model for a Rectenna and Its Application to an RF Harvester with MPPT.

This paper proposes a compact Thévenin model for a rectenna. This model is then applied to design a high-efficiency radio frequency harvester with a maximum power point tracker (MPPT). The rectenna under study consists of an L-matching network and a half-wave rectifier. The derived model is simpler and more compact than those suggested so far in the literature and includes explicit expressions of the Thévenin voltage (Voc) and resistance and of the power efficiency related with the parameters of the rectenna. The rectenna was implemented and characterized from −30 to −10 dBm at 808 MHz. Experimental results agree with the proposed model, showing a linear current–voltage relationship as well as a maximum efficiency at Voc/2, in particular 60% at −10 dBm, which is a remarkable value. An MPPT was also used at the rectenna output in order to automatically work at the maximum efficiency point, with an overall efficiency near 50% at −10 dBm. Further tests were performed using a nearby transmitting antenna for powering a sensor node with a power consumption of 4.2 µW.

Preferential generation of Ca2+-permeable AMPA receptors by AKAP79-anchored protein kinase C proceeds via GluA1 subunit phosphorylation at Ser-831

AMPA-type glutamate receptors (AMPARs) mediate fast excitatory neurotransmission in the mammalian central nervous system. Preferential AMPAR subunit assembly favors heteromeric GluA1/GluA2 complexes. The presence of the GluA2 subunit generates Ca2+-impermeable (CI) AMPARs that have linear current-voltage (I-V) relationships. However, diverse forms of synaptic plasticity and pathophysiological conditions are associated with shifts from CI to inwardly rectifying, GluA2-lacking, Ca2+-permeable (CP) AMPARs on time scales ranging from minutes to days. These shifts have been linked to GluA1 phosphorylation at Ser-845, a protein kinase A (PKA)-targeted site within its intracellular C-terminal tail, often in conjunction with protein kinase A anchoring protein 79 (AKAP79; AKAP150 in rodents), which targets PKA to GluA1. However, AKAP79 may impact GluA1 phosphorylation at other sites by interacting with other signaling enzymes. Here, we evaluated the ability of AKAP79, its signaling components, and GluA1 phosphorylation sites to induce CP-AMPARs under conditions in which CI-AMPARs normally predominate. We found that GluA1 phosphorylation at Ser-831 is sufficient for the appearance of CP-AMPARs and that AKAP79-anchored protein kinase C (PKC) primarily drives the appearance of these receptors via this site. In contrast, other AKAP79-signaling components and C-terminal tail GluA1 phosphorylation sites exhibited a permissive role, limiting the extent to which AKAP79 promotes CP-AMPARs. This may reflect the need for these sites to undergo active phosphorylation/dephosphorylation cycles that control their residency within distinct subcellular compartments. These findings suggest that AKAP79, by orchestrating phosphorylation, represents a key to a GluA1 phosphorylation passcode, which allows the GluA1 subunit to escape GluA2 dominance and promote the appearance of CP-AMPARs.

Piezo2 Mechanosensitive Ion Channel Role in Primary Enterochromaffin (EC) Cell Mechanosensitivity

INTRODUCTIONIntestinal enterochromaffin (EC) cells are the specialized epithelial mechanosensors of the gastrointestinal (GI) epithelium and are functionally and developmentally like Merkel cells in the skin. EC cells release serotonin (5‐HT) in response to mechanical stimuli. Recently, we discovered that like Merkel cells, human and mouse EC cells express the mechanosensitive (MS) ion channel Piezo2. However, little is known about the functional properties of this channel in primary EC cells.AIMTo determine the role of Piezo2 in primary EC cells.METHODSThe enteroendocrine cell transcription factor NeuroD1 was used to mark the late stage of EC cell development and generate NeuroD1‐cre::GCaMP5/tdTomato mouse to identify EC cells and examine their Ca2+ dynamics. We examined colocalization of tdTomato with 5‐HT and Piezo2 by immunohistochemistry. We established and used primary colon epithelium cultures for whole‐cell patch clamp and Ca2+ imaging experiments and mechanically stimulated by membrane displacement with a piezoelectrically driven glass probe or shear flow.RESULTSImmunostaining showed no tdTomato labeling outside of the GI epithelium, and epithelial tdTomato cells were 5‐HT+ and Piezo2+. Mechanical stimulation of primary EC cells by membrane displacement produced inward MS currents (20.8±4.9 pA/pF, Cm=3.5 pF, n=6) with fast activation and inactivation kinetics (τinact 11.1±2.8 ms), steep mechanosensitivity (Boltzmann function fit with a mid‐point of 3.46±0.80 μm and slope 0.79±0.27 μm) and a linear current‐voltage relationship (slope 0.61±0.17 pA/mV, x‐intercept −4.26±1.34 mV, n=5). Shear flow (20 s) transiently increased intracellular calcium ((Ca2+)i) (2.15±0.5 ΔF/F0, upstroke 3–5 s, return to baseline in 60 s, n=12). MS currents were inhibited by the MS ion channel blocker Gd3+ (20.8±4.9 pA/pF versus 5.3±1.9 pA/pF), Piezo channel blocker GsMTx‐4 (20.8±4.9 pA/pF versus 2.5±0.9 pA/pF) and Piezo2 siRNA (20.8±4.9 pA/pF versus 1.1±0.3 pA/pF), but not non‐targeted (NT) siRNA (20.8±4.9 pA/pF versus 16.0±5.2 pA/pF and) (n=4–6 for each condition, p<0.05 for all except NT siRNA). Similarly, the (Ca2+)i responses to force were inhibited by Gd3+ (2.28±0.7 ΔF/F0 to 0.09±0.05 ΔF/F0), GsMTx‐4 (2.0±0.6 ΔF/F0 to 0.3±0.1 ΔF/F0), Piezo2 siRNA (2.15±0.5 ΔF/F0 to 0.08±0.05 ΔF/F0), but not NT siRNA (2.15±0.5 ΔF/F0 to 2.7±0.3 ΔF/F0) (n=4–6, p<0.05 for all except NT siRNA).CONCLUSIONSIn the mouse epithelium, Piezo2+ EC cells were specifically marked by NeuroD1. Primary colonic NeuroD1+ cells had inward and non‐rectifying MS currents that could be blocked by Gd3+, GsMTx‐4 and Piezo2 siRNA. Similarly, shear flow evoked changes in (Ca2+)i that were also blocked by Piezo2 inhibitors and dependent on extracellular Ca2+. These results suggest that the MS ion channel Piezo2 acts as the primary EC cell mechanosensor, whose activation by force leads to MS ionic current and (Ca2+)i increase that may mediate mechanically‐induced 5‐HT release.Support or Funding InformationThis work was supported by NIH K08 DK106456, AGA Research Scholar Award, Mayo Clinic Center for Biomedical Discovery, Mayo Clinic Center for Cell Signaling in Gastroenterology (P30DK084567) and DK52766.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Plasticity of calcium-permeable AMPA glutamate receptors in Pro-opiomelanocortin neurons.

POMC neurons integrate metabolic signals from the periphery. Here, we show in mice that food deprivation induces a linear current-voltage relationship of AMPAR-mediated excitatory postsynaptic currents (EPSCs) in POMC neurons. Inhibition of EPSCs by IEM-1460, an antagonist of calcium-permeable (Cp) AMPARs, diminished EPSC amplitude in the fed but not in the fasted state, suggesting entry of GluR2 subunits into the AMPA receptor complex during food deprivation. Accordingly, removal of extracellular calcium from ACSF decreased the amplitude of mEPSCs in the fed but not the fasted state. Ten days of high-fat diet exposure, which was accompanied by elevated leptin levels and increased POMC neuronal activity, resulted in increased expression of Cp-AMPARs on POMC neurons. Altogether, our results show that entry of calcium via Cp-AMPARs is inherent to activation of POMC neurons, which may underlie a vulnerability of these neurons to calcium overload while activated in a sustained manner during over-nutrition.

Activation of a Ca2+-dependent cation conductance with properties of TRPM2 by reactive oxygen species in lens epithelial cells

Ion channels are crucial for maintenance of ion homeostasis and transparency of the lens. The lens epithelium is the metabolically and electrophysiologically active cell type providing nutrients, ions and water to the lens fiber cells. Ca2+-dependent non-selective ion channels seem to play an important role for ion homeostasis. The aim of the study was to identify and characterize Ca2+- and reactive oxygen species (ROS)-dependent non-selective cation channels in human lens epithelial cells. RT-PCR revealed gene expression of the Ca2+-activated non-selective cation channels TRPC3, TRPM2, TRPM4 and Ano6 in both primary lens epithelial cells and the cell line HLE-B3, whereas TRPM5 mRNA was only found in HLE-B3 cells. Using whole-cell patch-clamp technique, ionomycin evoked non-selective cation currents with linear current-voltage relationship in both cell types. The current was decreased by flufenamic acid (FFA), 2-APB, 9-phenanthrol and miconazole, but insensitive to DIDS, ruthenium red, and intracellularly applied spermine. H2O2 evoked a comparable current, abolished by FFA. TRPM2 protein expression in HLE-B3 cells was confirmed by means of immunocytochemistry and western blot. In summary, we conclude that lens epithelial cells functionally express Ca2+- and H2O2-activated non-selective cation channels with properties of TRPM2.

Contributions of space-clamp errors to apparent time-dependent loss of Mg2+ block induced by NMDA.

N-methyl-d-aspartate receptors (NMDARs) govern synaptic plasticity, development, and neuronal response to insult. Prolonged activation of NMDARs such as during an insult may activate secondary currents or modulate Mg2+ sensitivity, but the conditions under which these occur are not fully defined. We reexamined the effect of prolonged NMDAR activation in juvenile mouse hippocampal slices. NMDA (10 μM) elicited current with the expected negative-slope conductance in the presence of 1.2 mM Mg2+ However, several minutes of continued NMDA exposure elicited additional inward current at -70 mV. A higher concentration of NMDA (100 µM) elicited the current more rapidly. The additional current was not dependent on Ca2+, network activity, or metabotropic NMDAR function and did not persist on agonist removal. Voltage ramps revealed no alteration of either reversal potential or NMDA-elicited conductance between -30 mV and +50 mV. The result was a more linear NMDA current-voltage relationship. The current linearization was also induced in interneurons and in mature dentate granule neurons but not immature dentate granule cells, dissociated cultured hippocampal neurons, or nucleated patches excised from CA1 pyramidal neurons. Comparative simulations of NMDA application to a CA1 pyramidal neuron and to a cultured neuron revealed that linearization can be explained by space-clamp errors arising from gradual recruitment of distal dendritic NMDARs. We conclude that persistent secondary currents do not strongly contribute to NMDAR responses in juvenile mouse hippocampus and careful discernment is needed to exclude contributions of clamp artifacts to apparent secondary currents.NEW & NOTEWORTHY We report that upon sustained activation of NMDARs in juvenile mouse hippocampal neurons there is apparent loss of Mg2+ block at negative membrane potentials. However, the phenomenon is explained by loss of dendritic voltage clamp, leading to a linear current-voltage relationship. Our results give a specific example of how spatial voltage errors in voltage-clamp recordings can readily be misinterpreted as biological modulation.

Evidence that Orai1 does not contribute to store-operated TRPC1 channels in vascular smooth muscle cells

ABSTRACTCa2+-permeable store-operated channels (SOCs) mediate Ca2+ entry pathways which are involved in many cellular functions such as contraction, growth, and proliferation. Prototypical SOCs are formed of Orai1 proteins and are activated by the endo/sarcoplasmic reticulum Ca2+ sensor stromal interaction molecule 1 (STIM1). There is considerable debate about whether canonical transient receptor potential 1 (TRPC1) proteins also form store-operated channels (SOCs), and if they do, is Orai1 involved. We recently showed that stimulation of TRPC1-based SOCs involves store depletion inducing STIM1-evoked Gαq/PLCβ1 activity in contractile vascular smooth muscle cells (VSMCs). Therefore the present work investigates the role of Orai1 in activation of TRPC1-based SOCs in freshly isolated mesenteric artery VSMCs from wild-type (WT) and Orai1−/− mice. Store-operated whole-cell and single channel currents recorded from WT and Orai1−/− VSMCs had similar properties, with relatively linear current-voltage relationships, reversal potentials of about +20mV, unitary conductances of about 2pS, and inhibition by anti-TRPC1 and anti-STIM1 antibodies. In Orai1−/− VSMCs, store depletion induced PLCβ1 activity measured with the fluorescent phosphatidylinositol 4,5-bisphosphate/inositol 1,4,5-trisphosphate biosensor GFP-PLCδ1-PH, which was prevented by knockdown of STIM1. In addition, in Orai1−/− VSMCs, store depletion induced translocation of STIM1 from within the cell to the plasma membrane where it formed STIM1-TRPC1 interactions at discrete puncta-like sites. These findings indicate that activation of TRPC1-based SOCs through a STIM1-activated PLCβ1 pathway are likely to occur independently of Orai1 proteins, providing evidence that TRPC1 channels form genuine SOCs in VSMCs with a contractile phenotype.

Reactive oxygen species increase neuronal excitability via activation of nonspecific cation channel in rat medullary dorsal horn neurons.

The caudal subnucleus of the spinal trigeminal nucleus (medullary dorsal horn; MDH) receives direct inputs from small diameter primary afferent fibers that predominantly transmit nociceptive information in the orofacial region. Recent studies indicate that reactive oxygen species (ROS) is involved in persistent pain, primarily through spinal mechanisms. In this study, we aimed to investigate the role of xanthine/xanthine oxidase (X/XO) system, a known generator of superoxide anion (O2·−), on membrane excitability in the rat MDH neurons. For this, we used patch clamp recording and confocal imaging. An application of X/XO (300 µM/30 mU) induced membrane depolarization and inward currents. When slices were pretreated with ROS scavengers, such as phenyl N-tert-butylnitrone (PBN), superoxide dismutase (SOD), and catalase, X/XO-induced responses decreased. Fluorescence intensity in the DCF-DA and DHE-loaded MDH cells increased on the application of X/XO. An anion channel blocker, 4,4-diisothiocyanatostilbene-2,2-disulfonic acid (DIDS), significantly decreased X/XO-induced depolarization. X/XO elicited an inward current associated with a linear current-voltage relationship that reversed near −40 mV. X/XO-induced depolarization reduced in the presence of La3+, a nonselective cation channel (NSCC) blocker, and by lowering the external sodium concentration, indicating that membrane depolarization and inward current are induced by influx of Na+ ions. In conclusion, X/XO-induced ROS modulate the membrane excitability of MDH neurons, which was related to the activation of NSCC.