Influx Of Na Research Articles

α-Latrotoxin Actions in the Absence of Extracellular Ca2+ Require Release of Stored Ca2+

α-Latrotoxin (αLTX) causes exhaustive release of neurotransmitters from nerve terminals in the absence of extracellular Ca2+ (Ca2+e). To investigate the mechanisms underlying this effect, we loaded mouse neuromuscular junctions with BAPTA-AM. This membrane-permeable Ca2+-chelator demonstrates that Ca2+e-independent effects of αLTX require an increase in cytosolic Ca2+ (Ca2+cyt). We also show that thapsigargin, which depletes Ca2+ stores, induces neurotransmitter release, but inhibits the effect of αLTX. We then studied αLTX’s effects on Ca2+cyt using neuroblastoma cells expressing signaling-capable or signaling-incapable variants of latrophilin-1, a G protein-coupled receptor of αLTX. Our results demonstrate that αLTX acts as a cation ionophore and a latrophilin agonist. In model cells at 0 Ca2+e, αLTX forms membrane pores and allows the influx of Na+; this reverses the Na+-Ca2+ exchanger, leading to the release of stored Ca2+ and inhibition of its extrusion. Concurrently, αLTX stimulates latrophilin signaling, which depletes a Ca2+ store and induces transient opening of Ca2+ channels in the plasmalemma that are sensitive to inhibitors of store-operated Ca2+ entry. These results indicate that Ca2+ release from intracellular stores and that Ca2+ influx through latrophilin-activated store-operated Ca2+ channels contributes to αLTX actions and may be involved in physiological control of neurotransmitter release at nerve terminals.

Overexpression of phosphatidylserine synthase IbPSS1 enhances salt tolerance by stimulating ethylene signaling-dependent lignin synthesis in sweetpotato roots

Phosphatidylserine (PS) is an important lipid signaling required for plant growth regulation and salt stress adaptation. However, how PS positively regulate plant salt tolerance is still largely unknown. In this study, IbPSS1-overexpressed sweetpotato plants that exhibited overproduction of PS was employed to explore the mechanisms underlying the PS stimulation of plant salt tolerance. The results revealed that the IbPSS1-overexpressed sweetpotato accumulated less Na+ in the stem and leaf tissues compared with the wild type plants. Proteomic profile of roots showed that lignin synthesis-related proteins over-accumulated in IbPSS1-overexpressed sweetpotato. Correspondingly, the lignin content was enhanced but the influx of Na + into the stele was significantly blocked in IbPSS1-overexpressed sweetpotato. The results further revealed that ethylene synthesis and signaling related genes were upregulated in IbPSS1-overexpressed sweetpotato. Ethylene imaging experiment revealed the enhancement of ethylene mainly localized in the root stele. Inhibition of ethylene synthesis completely reversed the PS-overproduction induced lignin synthesis and Na+ influx pattern in stele tissues. Taken together, our findings demonstrate a mechanism by which PS regulates ethylene signaling and lignin synthesis in the root stele, thus helping sweetpotato plants to block the loading of Na+ into the xylem and to minimize the accumulation of Na+ in the shoots.

Theoretical calculation of NMDA receptor desensitization and intracellular calcium determination through simulation

One of the crucial processes in preventing cellular damage due to excitotoxicity involves the calcium-mediated desensitization of the N-methyl-D-aspartate (NMDA) receptor. The opening of NMDA channels results in the influx of Na+ and Ca2+ ions and efflux of K+ ions. This condition leads to an increase in intracellular calcium ([Ca2+]i), thereby diminishing the NMDA current and preventing damage induced by constant stimulation of the receptor by glutamate. In this study, a simulator based on a phenomenological mathematical model was developed, employing the Visual Basic 6.0 language for the Windows® environment. Input variables encompass intracellular calcium, peak currents (Ip), and steady-state currents (Iss). Input currents were derived from previously published electrophysiological recordings. The simulator adeptly reproduces the NMDA receptor desensitization phenomenon under varying experimental conditions and accurately calculates internal calcium with a 10% margin of error. It proves valuable in scenarios where simultaneous recordings of NMDA current and [Ca2+]i via fluorescence labeling techniques are impractical. In the realm of education, it enables students to conduct virtual experiments, providing an immersive introduction to the topic.

Membrane procoagulation and N‑terminomics/TAILS profiling in Montreal platelet syndrome kindred with VWF p.V1316M mutation

BackgroundThe Montreal platelet syndrome kindred (MPS) with VWF p.V1316M mutation (2B-VWDMPS) is an extremely rare disorder. It has been associated with macrothrombocytopenia, spontaneous platelet clumping, mucocutaneous, and other bleeding, which can be largely prevented by von Willebrand factor (VWF) concentrate infusion. However, supplemental platelet transfusion has been required on occasion, particularly for severe gastrointestinal bleeds. This raised the question of whether a previously uncharacterized platelet dysfunction contributes to bleeding diathesis in 2B-VWDMPS patients. We have previously shown that membrane ballooning, a principal part of the platelet procoagulant membrane dynamics (PMD) after collagen stimulation, is driven by the influx of Na+ and Cl-, followed by the entry of water.MethodsWe study two members (mother and daughter) of the MPS kindred with severe bleeding phenotype and address this question by coupling quantitative platelet shotgun proteomics and validating biochemical assays, with the systematic analysis of platelet procoagulant membrane dynamics (PMD). Using N-terminomics/TAILS (terminal amine isotopic labeling of substrates), we compare changes in proteolysis between healthy and 2B-VWDMPS platelets.ResultsHere, we report in 2B-VWDMPS platelets, the loss of the transmembrane chloride channel-1 (CLIC1), and reduced chloride ion influx after collagen stimulation. This was associated with diminished membrane ballooning, phosphatidylserine externalization, and membrane thrombin formation, as well as a distinct phenotypic composition of platelets over fibrillar collagen. We also identify processing differences of VWF, fibronectin (FN1), and Crk-like protein (CRKL). 2B-VWDMPS platelets are shown to be basally activated, partially degranulated, and have marked loss of regulatory, cytoskeletal, and contractile proteins.ConclusionsThis may account for structural disorganization, giant platelet formation, and a weakened hemostatic response.

Generation of a TRPV1 knockout human pluripotent stem cell line (WAe009-A-U) using CRISPR/Cas9

The transient receptor potential vanilloid subfamily 1 (TRPV1) is a polymodal nociceptor that is highly expressed in sensory nerves. Activation of TRPV1 receptors excites primary afferent nociceptors by opening cation channels, allowing the influx of Na+ and Ca2+ ions into the cytoplasm. Here, a TRPV1 knockout human embryonic stem cell line was generated using the CRISPR/Cas9 genome-editing technology to further study the function of TRPV1. The cell line confirmed with normal pluripotency and karyotype.

Cation flux through SUR1-TRPM4 and NCX1 in astrocyte endfeet induces water influx through AQP4 and brain swelling after ischemic stroke.

Brain swelling causes morbidity and mortality in various brain injuries and diseases but lacks effective treatments. Brain swelling is linked to the influx of water into perivascular astrocytes through channels called aquaporins. Water accumulation in astrocytes increases their volume, which contributes to brain swelling. Using a mouse model of severe ischemic stroke, we identified a potentially targetable mechanism that promoted the cell surface localization of aquaporin 4 (AQP4) in perivascular astrocytic endfeet, which completely ensheathe the brain's capillaries. Cerebral ischemia increased the abundance of the heteromeric cation channel SUR1-TRPM4 and of the Na+/Ca2+ exchanger NCX1 in the endfeet of perivascular astrocytes. The influx of Na+ through SUR1-TRPM4 induced Ca2+ transport into cells through NCX1 operating in reverse mode, thus raising the intra-endfoot concentration of Ca2+. This increase in Ca2+ stimulated calmodulin-dependent translocation of AQP4 to the plasma membrane and water influx, which led to cellular edema and brain swelling. Pharmacological inhibition or astrocyte-specific deletion of SUR1-TRPM4 or NCX1 reduced brain swelling and improved neurological function in mice to a similar extent as an AQP4 inhibitor and was independent of infarct size. Thus, channels in astrocyte endfeet could be targeted to reduce postischemic brain swelling in stroke patients.

CHLOROGENIC ACID CAN INHIBIT CATECHOLAMINE SECRETION FROM THE ADRENAL MEDULLA

Objective: Chlorogenic acid (CGA) is one of the most available phenolic acid compounds in foods, such as coffee and tea, which is known to be the most effective agent in lowering blood pressure and helpful in treating hypertension-related disorders. Thus the present study was aimed to determine the characteristics of CGA on secretion of catecholamines (CA) in the perfused model of the rat adrenal medulla, and also to validify its mechanism of action. Design and method: he adrenal gland was isolated and perfused with Krebs-bicarbonate. CA was measured directly by using the fluorospectrophotometer. Results: CGA, administered into an adrenal vein for 90 min, reduced ACh -induced CA secretion in a dose- and time-dependent manner. CGA also time-dependently depressed the CA secretion induced by McN-A-343, DMPP, and angiotensin II . CGA itself failed to influence spontaneous CA secretion. Also, During the perfusion of CGA for 90 min, the CA secretory responses induced by high K+, veratridine, cyclopiazonic acid, Bay-K-8644 were suppressed. However, in the simultaneous presence of CGA and L-NAME, the CA secretory responses induced by ACh, Ang II, Bay-K-8644 and veratridine was restored nearly to the control level compared to those of the inhibitory effects of CGA-treatment alone. Virtually, the adrenal NO release was significantly enhanced by CGA-treatment. Also, in the coexistence of CGA and olmesartan, ACh-induced CA secretion was markedly reduced compared to that of olmesartan-treatment alone. Conclusions: Taken together, we present the first evidence that CGA suppresses the CA secretion induced by activation of cholinoceptors as well as AT1-receptors. This CGA-induced inhibitory effect seems to be exerted by reducing both the influx of Na+ and Ca2+ through their ionic channels into adrenomedullary chromaffin cells as well as by suppressing the Ca2+ release from the cytoplasmic calcium store, through the elevated NO release by the activation of NO synthase, which is associated to the blockade of neuronal cholinonoceptors and AT1-receptors. Based on these results, the ingestion of CGA may be helpful to alleviate or prevent the cardiovascular diseases, via reduction of adrenal CA secretion and consequent decreased CA level in circulation.

Functional modulation of a pH-sensitive ion channel by a transporter family.

The acid-sensing ion channel 1a (ASIC1a) is a trimeric ligand-gated ion channel. Its activation by protons results in influx of Na+ and, to a lesser extent, Ca2+ before the channel rapidly desensitizes. ASIC1a-mediated currents contribute to normal brain function, including long term potentiation, which is important for learning and memory. However, elevated activity of ASIC1a has been directly implicated in the pathogenesis of numerous disorders, such as ischemic brain injury, cancer, pain and Alzheimer's disease. Recent evidence suggests that during such pathological conditions, association with protein interaction partners can affect ASIC1a activity, thereby contributing to neuronal damage and cell death. Consequently, there is an increasing interest in understanding the regulation of ASIC1a by protein-interaction partners, which remain largely enigmatic. Here, we have used a combination of mass spectrometry, biochemistry and electrophysiology to identify a family of transporters that directly interacts with ASIC1a and regulates its function. Depending on the specific transporter isoform present in the channel-transporter complex, the activation of ASIC1a is either prolonged and potentiated or decreased. We anticipate this work to offer promising starting points for therapeutic targeting of ASIC1a-containing macromolecular complexes in various diseases.

S-13-4: INHIBITORY IMPACTS OF CHLOROGENIC ACID ON ADRENAL CATECHOLAMINE SECRETION

(Objective) Chlorogenic acid (CGA) is one of the most available phenolic acid compounds in foods, such as coffee and tea. Thus, the present study was aimed to determine the characteristics of CGA on secretion of catecholamines (CA) in the perfused model of the rat adrenal medulla, and also to validify its mechanism of action. (Design and Methods) The adrenal gland was isolated and perfused with Krebs-bicarbonate. CA was measured directly by using the fluorospectrophotometer. (Results) CGA (30∼300 μM), administered into an adrenal vein for 90 min, reduced ACh-induced CA secretion in a dose- and time-dependent manner. CGA also time-dependently depressed the CA secretion induced by McN-A-343 (a selective muscarinic M1 receptor agonist), DMPP (a selective neuronal nicotinic receptor agonist), and angiotensin II. CGA itself failed to influence spontaneous CA secretion. Also, During the perfusion of CGA for 90 min, the CA secretory responses induced by high K+ (56 mM, a direct membrane depolarizer), veratridine (a voltage-dependent Na+ channel activator, cyclopiazonic acid (a cytoplasmic Ca2+-ATPase inhibitor, Bay-K-8644 (an L-type dihydropyridine Ca2+ channel activator were suppressed. However, in the simultaneous presence of CGA and L-NAME (an inhibitor of NO synthase), the CA secretory responses induced by ACh, Ang II, Bay-K-8644 and veratridine was restored nearly to the control level compared to those of the inhibitory effects of CGA-treatment alone. Virtually, the adrenal NO release was significantly enhanced by CGA-treatment. Also, in the coexistence of CGA and olmesartan, ACh-induced CA secretion was markedly reduced compared to that of olmesartan-treatment alone. (Conclusion) Taken together, we present the first evidence that CGA suppresses the CA secretion induced by activation of cholinergic receptors as well as AT1-receptors. This CGA-induced inhibitory effect seems to be exerted by reducing both the influx of Na+ and Ca2+ through their ionic channels into the adrenomedullary chromaffin cells as well as by suppressing the Ca2+ release from the cytoplasmic calcium store, at least through the elevated NO release by the activation of NO synthase, which is associated to the blockade of neuronal cholinergic receptors and AT1-receptors. Based on these results, the ingestion of CGA may be helpful to alleviate or prevent the cardiovascular diseases, via reduction of adrenal CA secretion and consequent decreased CA level in the circulation. Co-administration of CGA and Olmesartan (ARB) may be clinically beneficial for treatment of cardiovascular diseases, including hypertension and angina pectoris.

“Virtual patch clamp analysis” for predicting the functional significance of pathogenic variants in sodium channels

ObjectiveOpening of voltage-gated sodium channels is crucial for neuronal depolarization. Proper channel opening and influx of Na+ through the ion pore, is dependent upon binding of Na+ ion to a specific amino-acid motif (DEKA) within the pore. In this study we used molecular dynamic simulations, an advanced bioinformatic tool, to research the dysfunction caused by pathogenic variants in SCN1a, SCN2a and SCN8a genes. MethodMolecular dynamic simulations were performed in six patients: three patients with Dravet syndrome (p.Gly177Ala,p.Ser259Arg and p.Met1267Ile, SCN1a), two patients with early onset drug resistant epilepsy(p.Ala263Val, SCN2a and p.Ile251Arg, SCN8a), and a patient with autism (p.Thr155Ala, SCN2a). After predicting the 3D-structure of mutated proteins by homology modeling, time dependent molecular dynamic simulations were performed, using the Schrödinger algorithm. The opening of the sodium channel, including the detachment of the sodium ion to the DEKA motif and pore diameter were assessed. Results were compared to the existent patch clamp analysis in four patients, and consistency with clinical phenotype was noted. ResultsThe Na+ ion remained attached to DEKA filter longer when compared to wild type in the p.Gly177Ala, p.Ser259Arg,SCN1a, and p.Thr155Ala, SCN2a variants, consistent with loss-of-function. In contrast, it detached quicker from DEKA than wild type in the p.Ala263Val,SCN2a variant, consistent with gain-of-function. In the p.Met1267Ile,SCN1a variant, detachment from DEKA was quicker, but pore diameter decreased, suggesting partial loss-of-function. In the p.Leu251Arg,SCN8a variant, the pore remained opened longer when compared to wild type, consistent with a gain-of-function. The molecular dynamic simulation results were consistent with the existing patch-clamp analysis studies, as well as the clinical phenotype. SignificanceMolecular dynamic simulation can be useful in predicting pathogenicity of variants and the disease phenotype, and selecting targeted treatment based on channel dysfunction. Further development of these bioinformatic tools may lead to “virtual patch-clamp analysis”.

Pretreating poplar cuttings with low nitrogen ameliorates salt stress responses by increasing stored carbohydrates and priming stress signaling pathways

Soil salinity is a widespread stress in semi-arid forests worldwide, but how to manage nitrogen (N) nutrition to improve plant saline tolerance remains unclear. Here, the cuttings of a widely distributed poplar from central Asia, Populus russikki Jabl., were exposed to either normal or low nitrogen (LN) concentrations for two weeks in semi-controlled greenhouse, and then they were added with moderate salt solution or not for another two weeks to evaluate their physiological, biochemical, metabolites and transcriptomic profile changes. LN-pretreating alleviated the toxicity caused by the subsequent salt stress in the poplar plants, demonstrated by a significant reduction in the influx of Na+ and Cl- and improvement of the K+/Na+ ratio. The other salt-stressed traits were also ameliarated, indicated by the variations of chlorophyll content, PSII photochemical activity and lipid peroxidation. Stress alleviation resulted from two different processes. First, LN pretreatment caused a significant increase of non-structural carbohydrates (NSC), allowed for an increased production of osmolytes and a higher potential fueling ion transport under subsequent salt condition, along with increased transcript levels of the cation/H+ ATPase. Second, LN pretreatment enhanced the transcript levels of stress signaling components and phytohormones pathway as well as antioxidant enzyme activities. The results indicate that early restrictions of N supply could enhance posterior survival under saline stress in poplar plants, which is important for plantation programs and restoration activities in semi-arid areas.

Slo2/KNa Channels in Drosophila Protect against Spontaneous and Induced Seizure-like Behavior Associated with an Increased Persistent Na+ Current.

Na+ sensitivity is a unique feature of Na+-activated K+ (KNa) channels, making them naturally suited to counter a sudden influx in Na+ ions. As such, it has long been suggested that KNa channels may serve a protective function against excessive excitation associated with neuronal injury and disease. This hypothesis, however, has remained largely untested. Here, we examine KNa channels encoded by the Drosophila Slo2 (dSlo2) gene in males and females. We show that dSlo2/KNa channels are selectively expressed in cholinergic neurons in the adult brain, as well as in glutamatergic motor neurons, where dampening excitation may function to inhibit global hyperactivity and seizure-like behavior. Indeed, we show that effects of feeding Drosophila a cholinergic agonist are exacerbated by the loss of dSlo2/KNa channels. Similar to mammalian Slo2/KNa channels, we show that dSlo2/KNa channels encode a TTX-sensitive K+ conductance, indicating that dSlo2/KNa channels can be activated by Na+ carried by voltage-dependent Na+ channels. We then tested the role of dSlo2/KNa channels in established genetic seizure models in which the voltage-dependent persistent Na+ current (INap) is elevated. We show that the absence of dSlo2/KNa channels increased susceptibility to mechanically induced seizure-like behavior. Similar results were observed in WT flies treated with veratridine, an enhancer of INap Finally, we show that loss of dSlo2/KNa channels in both genetic and pharmacologically primed seizure models resulted in the appearance of spontaneous seizures. Together, our results support a model in which dSlo2/KNa channels, activated by neuronal overexcitation, contribute to a protective threshold to suppress the induction of seizure-like activity.SIGNIFICANCE STATEMENT Slo2/KNa channels are unique in that they constitute a repolarizing K+ pore that is activated by the depolarizing Na+ ion, making them naturally suited to function as a protective "brake" against overexcitation and Na+ overload. Here, we test this hypothesis in vivo by examining how a null mutation of the Drosophila Slo2 (dSlo2)/KNa gene affects seizure-like behavior in genetic and pharmacological models of epilepsy. We show that indeed the loss of dSlo2/KNa channels results in increased incidence and severity of induced seizure behavior, as well as the appearance of spontaneous seizure activity. Our results advance our understanding of neuronal excitability and protective mechanisms that preserve normal physiology and the suppression of seizure susceptibility.

Social transmission in the wild can reduce predation pressure on novel prey signals

Social transmission of information is taxonomically widespread and could have profound effects on the ecological and evolutionary dynamics of animal communities. Demonstrating this in the wild, however, has been challenging. Here we show by field experiment that social transmission among predators can shape how selection acts on prey defences. Using artificial prey and a novel approach in statistical analyses of social networks, we find that blue tit (Cyanistes caeruleus) and great tit (Parus major) predators learn about prey defences by watching others. This shifts population preferences rapidly to match changes in prey profitability, and reduces predation pressure from naïve predators. Our results may help resolve how costly prey defences are maintained despite influxes of naïve juvenile predators, and suggest that accounting for social transmission is essential if we are to understand coevolutionary processes.

Palmitoylation Controls NMDA Receptor Function and Steroid Sensitivity.

NMDARs are ligand-gated ion channels that cause an influx of Na+ and Ca2+ into postsynaptic neurons. The resulting intracellular Ca2+ transient triggers synaptic plasticity. When prolonged, it may induce excitotoxicity, but it may also activate negative feedback to control the activity of NMDARs. Here, we report that a transient rise in intracellular Ca2+ (Ca2+ challenge) increases the sensitivity of NMDARs but not AMPARs/kainate receptors to the endogenous inhibitory neurosteroid 20-oxo-5β-pregnan-3α-yl 3-sulfate and to its synthetic analogs, such as 20-oxo-5β-pregnan-3α-yl 3-hemipimelate (PAhPim). In cultured hippocampal neurons, 30 μm PAhPim had virtually no effect on NMDAR responses; however, following the Ca2+ challenge, it inhibited the responses by 62%; similarly, the Ca2+ challenge induced a 3.7-fold decrease in the steroid IC50 on recombinant GluN1/GluN2B receptors. The increase in the NMDAR sensitivity to PAhPim was dependent on three cysteines (C849, C854, and C871) located in the carboxy-terminal domain of the GluN2B subunit, previously identified to be palmitoylated (Hayashi et al., 2009). Our experiments suggested that the Ca2+ challenge induced receptor depalmitoylation, and single-channel analysis revealed that this was accompanied by a 55% reduction in the probability of channel opening. Results of in silico modeling indicate that receptor palmitoylation promotes anchoring of the GluN2B subunit carboxy-terminal domain to the plasma membrane and facilitates channel opening. Depalmitoylation-induced changes in the NMDAR pharmacology explain the neuroprotective effect of PAhPim on NMDA-induced excitotoxicity. We propose that palmitoylation-dependent changes in the NMDAR sensitivity to steroids serve as an acute endogenous mechanism that controls NMDAR activity.SIGNIFICANCE STATEMENT There is considerable interest in negative allosteric modulators of NMDARs that could compensate for receptor overactivation by glutamate or de novo gain-of-function mutations in neurodevelopmental disorders. By a combination of electrophysiological, pharmacological, and computational techniques we describe a novel feedback mechanism regulating NMDAR activity. We find that a transient rise in intracellular Ca2+ increases NMDAR sensitivity to inhibitory neurosteroids in a process dependent on GluN2B subunit depalmitoylation. These results improve our understanding of the molecular mechanisms of steroid action at the NMDAR and indeed of the basic properties of this important glutamate-gated ion channel and may aid in the development of therapeutics for treating neurologic and psychiatric diseases related to overactivation of NMDARs without affecting normal physiological functions.

Continuous monitoring of plant sodium transport dynamics using clinical PET

BackgroundThe absorption, translocation, accumulation and excretion of substances are fundamental processes in all organisms including plants, and have been successfully studied using radiotracers labelled with 11C, 13N, 14C and 22Na since 1939. Sodium is one of the most damaging ions to the growth and productivity of crops. Due to the significance of understanding sodium transport in plants, a significant number of studies have been carried out to examine sodium influx, compartmentation, and efflux using 22Na- or 24Na-labeled salts. Notably, however, most of these studies employed destructive methods, which has limited our understanding of sodium flux and distribution characteristics in real time, in live plants. Positron emission tomography (PET) has been used successfully in medical research and diagnosis for decades. Due to its ability to visualise and assess physiological and metabolic function, PET imaging has also begun to be employed in plant research. Here, we report the use of a clinical PET scanner with a 22Na tracer to examine 22Na-influx dynamics in barley plants (Hordeum vulgare L. spp. Vulgare—cultivar Bass) under variable nutrient levels, alterations in the day/night light cycle, and the presence of sodium channel inhibitors.Results3D dynamic PET images of whole plants show readily visible 22Na translocation from roots to shoots in each examined plant, with rates influenced by both nutrient status and channel inhibition. PET images show that plants cultivated in low-nutrient media transport more 22Na than plants cultivated in high-nutrient media, and that 22Na uptake is suppressed in the presence of a cation-channel inhibitor. A distinct diurnal pattern of 22Na influx was discernible in curves displaying rates of change of relative radioactivity. Plants were found to absorb more 22Na during the light period, and anticipate the change in the light/dark cycle by adjusting the sodium influx rate downward in the dark period, an effect not previously described experimentally.ConclusionsWe demonstrate the utility of clinical PET/CT scanners for real-time monitoring of the temporal dynamics of sodium transport in plants. The effects of nutrient deprivation and of ion channel inhibition on sodium influx into barley plants are shown in two proof-of-concept experiments, along with the first-ever 3D-imaging of the light and dark sodium uptake cycles in plants. This method carries significant potential for plant biology research and, in particular, in the context of genetic and treatment effects on sodium acquisition and toxicity in plants.

Conductance stability and Na+ interaction with Shab K+ channels under low K+ conditions

ABSTRACT K+ ions exert a structural effect that brings stability to K+ selective pores. Thus, upon bathing Shab channels in 0 K+ solutions the ion conductance, GK, irreversibly collapses. Related to this, studies with isolated KcsA channels have suggested that there is a transition [K+] around which the pore takes one of two conformations, either the low (non-conducting) or high K+ (conducting) crystal structures. We examined this premise by looking at the K+-dependency of GK stability of Shab channels within the cell membrane environment. We found that: K+ effect on GK stability is highly asymmetrical, and that as internal K+ is replaced by Na+ GK drops in a way that suggests a transition internal [K+]. Additionally, we found that external permeant ions inhibit GK drop with a potency that differs from the global selectivity-sequence of K+ pores; the non-permeant TEA inhibited GK drop in a K+-dependent manner. Upon lowering internal [K+] we observed an influx of Na+ at negative potentials. Na+ influx was halted by physiological external [K+], which also restored GK stability. Hyperpolarized potentials afforded GK stability but, as expected, do not restore GK selectivity. For completeness, Na+ interaction with Shab was also assessed at depolarized potentials by looking at Na block followed by permeation (pore unblock) at positive potentials, in solutions approaching the 0 K+ limit. The stabilizing effect of negative potentials along with the non-parallel variation of Na+ permeability and conductance-stability herein reported, show that pore stability and selectivity, although related, are not strictly coupled.

Drainage of subduction interface fluids into the forearc mantle evidenced by a pristine jadeitite network (Polar Urals)

AbstractThe physical and mechanical processes rooted in the hydrated, serpentinized mantle above subduction zones remain insufficiently explored despite fundamental implications for our understanding of rheology and fluid recycling along subduction interfaces. Through a field‐based investigation, serpentinized peridotites and jadeitite samples from a fossil forearc mantle in the Polar Urals (Russia) are studied here to document fluid–rock interaction processes in the high‐P field, as well as the long‐term evolution of the base of the mantle wedge. Petrographic, geochemical and microstructural observations reveal a complex, protracted evolution of the jadeitite‐forming fluid pathway throughout the gradual cooling of the forearc mantle and increasing serpentinization of the host. It is shown that the jadeitite lenses in the studied locality (a) derive for a large part from a trondhjemitic dyke earlier emplaced in a warm subduction environment, and (b) record the cooling of the subduction hangingwall under high‐P conditions associated with increasing host serpentinization. In the studied locality, the majority of the jadeitites formed at relatively high temperatures (>600°C) by the influx of Na–Al‐rich, slab‐derived metamorphic fluids that were drained along the base of the mantle wedge, parallel to the subduction interface. Changes in bulk‐rock geochemical signatures and in paragenetic sequences also constrain the compositional evolution of the fluid channelized along this drainage, with an increasing sedimentary component. The phlogopite‐bearing walls of the dyke exhibit Rb–Sr and Ar–Ar ages ranging between c. 405 and c. 390 Ma, a range partly overlapping within uncertainty with the previously dated zircons from the jadeitite core (410–⁠400 Ma; U–Pb). This study opens a unique window on the pristine structures formed above the plate interface by melting and fluid–rock interaction in the early subduction stages, as well as their evolution during secular cooling of the base of the mantle wedge.

CaMKII delta interaction with neuronal sodium channel Nav1.8 contributes to arrhythmogenic triggers in failing human and mouse cardiomyocytes

Abstract In heart failure, enhanced persistent current through neuronal sodium channel NaV1.8 (INaL) may induce influx of Na+ into cardiomyocytes. This may cause Ca2+ influx via the Na+/Ca2+ exchanger leading to increased proarrhythmogenic diastolic sarcoplasmic reticulum (SR) Ca2+ leak. This Ca2+ may activate Ca2+/calmodulin-dependent protein kinase IIδ (CaMKIIδ) which can induce INaL augmentation by phosphorylating NaV1.5 channels leading to a vicious cycle between INaL and CaMKIIδ. Here, we examined whether CaMKIIδ associates with NaV1.8 in human and mouse cardiomyocytes thereby regulating its function. Interaction and co-localisation of CaMKIIδ and NaV1.8 were confirmed by co-immunoprecipitation and immunocytochemistry. Whole-cell patch clamp showed a potent reduction of INaL after addition of novel specific Nav1.8 blockers, either A-803467 (30 nmol/L) or PF-01247324 (1 μmol/L) in failing mouse cardiomyocytes overexpressing CaMKIIδc (CaMKIIδc+/T: −109.4±10.6 vs A-803467: −56.9±11.7 and PF-01247324:−-69.9±8.6 A*ms*F-1). In failing human cardiomyocytes inhibition of either NaV1.8 or CaMKIIδ using AIP (1 μmol/L) or AIP and PF-01247324 together led to a significant and comparable decrease of INaL (control: −93.7±7.1 vs PF-01247324: −56.8±6.6; AIP: −44.2±6.6; AIP+PF-01247324: −39.8±5.4 A*ms*F-1). Furthermore, to confirm whether observed alterations in INaL after inhibition of NaV1.8 are not due to an overall reduction in peak sodium current (INa) we measured INa properties in mouse cardiomyocytes. Importantly, we observed no difference neither in the peak nor in inactivation between wild type (WT), WT with PF-01247324 and in mice lacking NaV1.8. Using confocal microscopy we investigated whether inhibition of the NaV1.8-mediated INaL could attenuate the increase of proarrhythmogenic SR Ca2+ spark frequency (CaSpF) caused by overexpression of CaMKIIδ in mice. We observed a significant reduction of CaSpF in both NaV1.8 inhibitor groups (PF-01247324: 0.51±0.08 and A-803467: 0.57±0.08 μm–1 s–1) compared to control (1.00±0.13 μm–1 s–1). Incubation of human failing cardiomyocytes with either AIP (0.35±0.06 μm–1 s–1) or PF-01247324 (0.44±0.11 μm–1 s–1), or blocking CaMKIIδ and NaV1.8 together (0.30±0.08 μm–1 s–1) resulted in significant decrease of CaSpF compared to control (0.89±0.13 μm–1 s–1). In conclusion, we show for the first time subcellular localisation of the neuronal sodium channel NaV1.8 and its interaction with CaMKIIδ in both human and mouse ventricular cardiomyocytes. Moreover, pharmacological inhibition of NaV1.8 caused a reduction of the augmented INaL and spontaneous diastolic SR-Ca2+ release in both failing human and mouse cardiomyocytes. NaV1.8 and CaMKIIδ interaction seem to play a relevant role for the generation of arrhythmogenic triggers (INaL & spontaneous diastolic SR-Ca2+ release) in both human and mouse cardiomyocytes from failing hearts. Funding Acknowledgement Type of funding source: None

Positive allosteric activation of glial EAAT-2 transporter protein: A novel strategy for Alzheimer’s disease

Excitatory amino acid transporter-2 (EAAT-2) protein localized in the membrane of glial cells are responsible for the clearance of glutamate in synapse and it plays a key role among the five glutamate transporters (EAATs) in regulating synaptic transmission and preventing excitotoxicity in neurons. EAAT-2 dysfunction has been associated with the neuropathology of Alzheimer’s disease (AD). Impairment of EAAT-2 transporter function results excess accumulation of glutamate in synaptic cleft that acts on post-synaptic glutaminergic receptors excessively resulting in influx of Na+ and Ca2+ ions into the neurons. This triggers excitotoxicity in post-synaptic neurons by activating apoptotic or necrotic pathways causing neurodegeneration in AD. The compounds that increase the EAAT-2 activity may have therapeutic potential for neuroprotection in AD. The positive allosteric site activation of EAAT-2 represents a promising entry point for the identification of novel pharmacological compounds for the management of neurodegenerative conditions involving glutamate-mediated excitotoxicity. We hypothesize, therefore, that the positive allosteric activators may enhance glutamate clearance from the synaptic cleft by promoting orthosteric binding of glutamate ligand in EAAT-2 transporter protein and attenuate the excitotoxicity in neurons and prevent the disease progression of AD.

Relationship between soil and groundwater salinity in the Western Canada Sedimentary Basin.

Saturated soil paste extracts indicate soluble ions in soil pore water that are available to vegetation. As such, they are thought to accurately describe the relationship between soil and groundwater salinity. To test this assumption, soil and groundwater samples were collected from 575 monitoring wells in saline regions of the Western Canadian Sedimentary Basin (WCSB). Samples were analyzed for electrical conductivity (EC) and Cl-, Na+, Ca2+, Mg2+, K+, SO42-, and HCO- 3 content. We compared groundwater ionic concentrations to paste extracts derived from matching soils, finding that differences from in situ soil porosity cause saturated pastes to underestimate groundwater salinity. Therefore, we provide pedotransfer functions for accurately calculating groundwater quality from soil data. In addition, we discuss the effects of porosity and soil composition on the saturated paste method, as measured through hydraulic conductivity, saturation percent, and sample lithology. Groundwater salinity may also influence further leaching of salts from soil. As produced water (NaCl brine) spills are common across the sulfate-rich soils of the WCSB, we considered the effects of NaCl on leaching of other ions, finding that influx of Na+ into groundwater is associated with increased sulfate leaching from soil. Therefore, considering the secondary effects of produced water on groundwater quality is essential to spill management.