Linear Current-voltage Relationship Research Articles

Activation of glutamate transport evokes rapid glutamine release from perisynaptic astrocytes

Stimulation of astrocytes by neuronal activity and the subsequent release of neuromodulators is thought to be an important regulator of synaptic communication. In this study we show that astrocytes juxtaposed to the glutamatergic calyx of Held synapse in the rat medial nucleus of the trapezoid body (MNTB) are stimulated by the activation of glutamate transporters and consequently release glutamine on a very rapid timescale. MNTB principal neurones express electrogenic system A glutamine transporters, and were exploited as glutamine sensors in this study. By simultaneous whole-cell voltage clamping astrocytes and neighbouring MNTB neurones in brainstem slices, we show that application of the excitatory amino acid transporter (EAAT) substrate d-aspartate stimulates astrocytes to rapidly release glutamine, which is detected by nearby MNTB neurones. This release is significantly reduced by the toxins L-methionine sulfoximine and fluoroacetate, which reduce glutamine concentrations specifically in glial cells. Similarly, glutamine release was also inhibited by localised inactivation of EAATs in individual astrocytes, using internal DL-threo-β-benzyloxyaspartic acid (TBOA) or dissipating the driving force by modifying the patch-pipette solution. These results demonstrate that astrocytes adjacent to glutamatergic synapses can release glutamine in a temporally precise, controlled manner in response to glial glutamate transporter activation. Since glutamine can be used by neurones as a precursor for glutamate and GABA synthesis, this represents a potential feedback mechanism by which astrocytes can respond to synaptic activation and react in a way that sustains or enhances further communication. This would therefore represent an additional manifestation of the tripartite relationship between synapses and astrocytes.

Erratum to: Mathematical properties of pump-leak models of cell volume control and electrolyte balance

Homeostatic control of cell volume and intracellular electrolyte content is a fundamental problem in physiology and is central to the functioning of epithelial systems. These physiological processes are modeled using pump-leak models, a system of differential algebraic equations that describes the balance of ions and water flowing across the cell membrane. Despite their widespread use, very little is known about their mathematical properties. Here, we establish analytical results on the existence and stability of steady states for a general class of pump-leak models. We treat two cases. When the ion channel currents have a linear current-voltage relationship, we show that there is at most one steady state, and that the steady state is globally asymptotically stable. If there are no steady states, the cell volume tends to infinity with time. When minimal assumptions are placed on the properties of ion channel currents, we show that there is an asymptotically stable steady state so long as the pump current is not too large. The key analytical tool is a free energy relation satisfied by a general class of pump-leak models, which can be used as a Lyapunov function to study stability.

Switch to glutamate receptor 2-lacking AMPA receptors increases neuronal excitability in hypothalamus and sympathetic drive in hypertension.

Glutamatergic synaptic input in the hypothalamic paraventricular nucleus (PVN) plays a critical role in regulating sympathetic outflow in hypertension. GluR2-lacking AMPA receptors (AMPARs) are permeable to Ca(2+), and their currents show unique inward rectification. However, little is known about changes in the AMPAR composition and its functional significance in hypertension. In this study, we found that AMPAR-mediated EPSCs (AMPAR-EPSCs) of retrogradely labeled spinally projecting PVN neurons exhibited a linear current-voltage relationship in Wistar-Kyoto (WKY) rats. However, AMPAR-EPSCs of labeled PVN neurons in spontaneously hypertensive rats (SHR) displayed inward rectification at positive holding potentials, which were not altered by lowering blood pressure with celiac ganglionectomy. Blocking GluR2-lacking AMPARs with 1-naphthyl acetyl spermine (NAS) caused a greater reduction in the AMPAR-EPSC amplitude and firing activity of PVN neurons in SHR than in WKY rats. Furthermore, blocking NMDA receptors and inhibition of calpain or calcineurin abolished inward rectification of AMPAR-EPSCs of PVN neurons in SHR. The GluR2 protein level was significantly less in the plasma membrane but greater in the cytosolic vesicle fraction in SHR than in WKY rats. In addition, microinjection of NAS into the PVN decreased blood pressure and lumbar sympathetic nerve activity in SHR but not in WKY rats. Our study reveals that increased GluR2-lacking AMPAR activity of PVN neurons results from GluR2 internalization through NMDA receptor-calpain-calcineurin signaling in hypertension. This phenotype switch in synaptic AMPARs contributes to increased excitability of PVN presympathetic neurons and sympathetic vasomotor tone in hypertension.

Enhanced current-rectification in bilayer graphene with an electrically tuned sloped bandgap

We propose a novel sloped dielectric geometry in graphene as a band engineering method for widening the depletion region and increasing the electrical rectification effect in graphene pn junctions. Enhanced current-rectification was achieved in a bilayer graphene with a sloped dielectric top gate and a normal back gate. A bias was applied to the top gate to induce a spatially modulated and sloped band configuration, while a back-gate bias was applied to open a bandgap. The sloped band can be tuned to separate n- and p-type regions in the bilayer graphene, depending on a suitable choice of gate voltage. The effective depletion region between the n- and p-type regions can be spatially enlarged due to the proposed top-gate structure. As a result, a strong non-linear electric current was observed during drain bias sweeping, demonstrating the expected rectification behavior with an on/off ratio higher than all previously reported values for graphene pn junctions. The observed rectification was modified to a linear current-voltage relationship by adjusting the biases of both gates to form an nn- or pp-type junction configuration. These results demonstrate that an external voltage can control the current flow in atomic film diodes.

A calcium-permeable non-selective cation channel in the thick ascending limb apical membrane of the mouse kidney

Non-selective cation channels have been described in the basolateral membrane of the renal tubule, but little is known about functional channels on the apical side. Apical membranes of microdissected fragments of mouse cortical thick ascending limbs were searched for ion channels using the cell-free configuration of the patch-clamp technique. A cation channel with a linear current–voltage relationship (19pS) that was permeable both to monovalent cations [PNH4(1.7)>PNa (1.0)=PK (1.0)] and to Ca2+ (PCa/PNa≈0.3) was detected. Unlike the basolateral TRPM4 Ca2+-impermeable non-selective cation channel, this non-selective cation channel was insensitive to internal Ca2+, pH and ATP. The channel was already active after patch excision, and its activity increased after reduced pressure was applied via the pipette. External gadolinium (10−5M) decreased the channel-open probability by 70% in outside-out patches, whereas external amiloride (10−4M) had no effect. Internal flufenamic acid (10−4M) inhibited the channel in inside-out patches. Its properties suggest that the current might be supported by the TRPM7 protein that is expressed in the loop of Henle. The conduction properties of the channel suggest that it could be involved in Ca2+ signaling.

Mathematical properties of pump-leak models of cell volume control and electrolyte balance

Homeostatic control of cell volume and intracellular electrolyte content is a fundamental problem in physiology and is central to the functioning of epithelial systems. These physiological processes are modeled using pump-leak models, a system of differential algebraic equations that describes the balance of ions and water flowing across the cell membrane. Despite their widespread use, very little is known about their mathematical properties. Here, we establish analytical results on the existence and stability of steady states for a general class of pump-leak models. We treat two cases. When the ion channel currents have a linear current-voltage relationship, we show that there is at most one steady state, and that the steady state is globally asymptotically stable. If there are no steady states, the cell volume tends to infinity with time. When minimal assumptions are placed on the properties of ion channel currents, we show that there is an asymptotically stable steady state so long as the pump current is not too large. The key analytical tool is a free energy relation satisfied by a general class of pump-leak models, which can be used as a Lyapunov function to study stability.

Sexual dimorphism and oestrogen regulation of KCNE3 expression modulates the functional properties of KCNQ1 K+ channels

The KCNQ1 potassium channel associates with various KCNE ancillary subunits that drastically affect channel gating and pharmacology. Co-assembly with KCNE3 produces a current with nearly instantaneous activation, some time-dependent activation at very positive potentials, a linear current-voltage relationship and a 10-fold higher sensitivity to chromanol 293B. KCNQ1:KCNE3 channels are expressed in colonic crypts and mediate basolateral K(+) recycling required for Cl(-) secretion. We have previously reported the female-specific anti-secretory effects of oestrogen via KCNQ1:KCNE3 channel inhibition in colonic crypts. This study was designed to determine whether sex and oestrogen regulate the expression and function of KCNQ1 and KCNE3 in rat distal colon. Colonic crypts were isolated from Sprague-Dawley rats and used for whole-cell patch-clamp and to extract total RNA and protein. Sheets of epithelium were used for short-circuit current recordings. KCNE1 and KCNE3 mRNA and protein abundance were significantly higher in male than female crypts. No expression of KCNE2 was found and no difference was observed in KCNQ1 expression between male and female (at oestrus) colonic crypts. Male crypts showed a 2.2-fold higher level of association of KCNQ1 and KCNE3 compared to female cells. In female colonic crypts, KCNQ1 and KCNE3 protein expression fluctuated throughout the oestrous cycle and 17β-oestradiol (E2 10 nM) produced a rapid (<15 min) dissociation of KCNQ1 and KCNE3 in female crypts only. Whole-cell K(+) currents showed a linear current-voltage relationship in male crypts, while K(+) currents in colonic crypts isolated from females displayed voltage-dependent outward rectification. Currents in isolated male crypts and epithelial sheets were 10-fold more sensitive to specific KCNQ1 inhibitors, such as chromanol 293B and HMR-1556, than in female. The effect of E2 on K(+) currents mediated by KCNQ1 with or without different β-subunits was assayed from current-voltage relations elicited in CHO cells transfected with KCNQ1 and KCNE3 or KCNE1 cDNA. E2 (100 nM) reduced the currents mediated by the KCNQ1:KCNE3 potassium channel and had no effect on currents via KCNQ1:KCNE1 or KCNQ1 alone. Currents mediated by the complex formed by KCNQ1 and the mutant KCNE3-S82A β-subunit (mutation of the site for PKCδ-promoted phosphorylation and modulation of the activity of KCNE3) showed rapid run-down and insensitivity to E2. Together, these data suggest that oestrogen regulates the expression of the KCNE1 and KCNE3 and with it the gating and pharmacological properties of the K(+) conductance required for Cl(-) secretion. The decreased association of the KCNQ1:KCNE3 channel complex promoted by oestrogen exposure underlies the molecular mechanism for the sexual dimorphism and oestrous cycle dependence of the anti-secretory actions of oestrogen in the intestine.

A self-consistent synthesis description of magnetosphere-ionosphere coupling and scale-dependent auroral process using shear Alfvén wave

[1] In order to correctly describe the dynamical behavior of the magnetosphere-ionosphere (MI) coupling system and the scale-dependent auroral process, we develop a synthesis formulation that combines the process of (1) the inverse Walen separation of MHD disturbance into parallel- and antiparallel-propagating shear Alfven wave to the ambient magnetic field, (2) the shear Alfven wave reflection process including (3) the scale-dependent electrostatic coupling process through the linearized Knight relation, (4) two-layer ionosphere model, and (5) dynamic conductance variations. A novel procedure that applies the inverse Walen relation to the incompressional MHD disturbances at the inner boundary of the MHD region enables to extract the component of the shear Alfven wave incident to the ionosphere. The extracted incident electric field supplies an electromotive force for the generation of the MI coupling system, and the reflected electric field is generated such that it totally satisfies the synthesis MI-coupling equation. A three-dimensional ionospheric current system is represented by a two-layer model in which the Pedersen and the Hall current are confined in the separated layers, which are connected by field-aligned currents driven by the linear current-voltage relation between two layers. Hence, our scheme possibly reproduces two types of the scale-dependent MI-decoupling process of the perpendicular potential structure: due to the parallel potential drop at the auroral acceleration region and the other due to the parallel potential differences inside the ionosphere. Our newly formulation may be well suited for description of scale-dependent auroral process and mesoscale ionospheric electrodynamics interlocked with the dynamical development of magnetospheric processes.

Ion channel activity of brain abundant protein BASP1 in planar lipid bilayers

BASP1 (also known as CAP-23 and NAP-22) is a brain abundant myristoylated protein localized at the inner surface of the presynaptic plasma membrane. Emerging evidence suggests that BASP1 is critically involved in various cellular processes, in particular, in the accumulation of phosphatidylinositol-4,5-diphosphate (PIP(2)) in lipid raft microdomains. We have recently shown that BASP1 forms heterogeneously-sized oligomers and higher aggregates with an outward similarity to oligomers and protofibrils of amyloid proteins. However, BASP1 is not known to be related to any amyloid disease. In the present study, we show that BASP1 induces single channel currents across negatively-charged planar lipid bilayers (containing phosphatidylserine or PIP(2)) bathed in 0.1-0.2 M KCl (pH 7.5). By their characteristics, BASP1 channels are similar to amyloid protein channels. BASP1 channels exhibit multiple conductance levels, in the range 10-3000 pS, with the most frequently observed conductance state of approximately 50 pS. The channels demonstrate a linear current-voltage relationship and voltage-independent kinetics of opening and closing. Their K(+) to Cl(-) permeability ratio is approximately 14, indicating that BASP1 channels are cation-selective. The ion channel activity of BASP1 is in accordance with the pore-like structure of BASP1 oligomers observed by electron microscopy on a lipid monolayer. Neuronal protein GAP-43, which is functionally related to BASP1 and also forms oligomers, elicited no ion channel currents under the conditions used in the present study. Elucidation of the physiological or pathological roles of ion channel activity of membrane-bound BASP1 oligomers will help to define the precise mechanism of amyloid protein toxicity.

Epithelial Barrier Resistance is Increased by the Divalent Cation Zinc in Cultured MDCKII Epithelial Monolayers

Topical zinc applications promote wound healing and epithelialization. "Leaky" MDCKII epithelia exposed to apical ZnCl₂ (10 mM) showed a time-dependent increase (t (0.5) 22.2 ± 2.7 min) of transepithelial resistance (R (t)) from 82.3 ± 2.4 Ω cm² to 1,551 ± 225.6 Ω cm²; the increase was dose-dependent, being observed at 3 mM but not at 1 mM. Basal Zn²+ applications also increased epithelial resistance (at 10 mM to 323 ± 225.6 Ω cm²). The linear current-voltage relationship in control epithelia changed after apical 10 mM ZnCl₂ to show rectification. Voltage deflections resulting from inward currents showed time-dependent relaxation (basal potential difference (p.d.)-positive), with outward currents being time-independent. Cation selectivity was tested after apical ZnCl₂ elevated resistance; both the NaCl:mannitol (basal replacement) dilution p.d. and the choline:Na bi-ionic p.d. decreased (P(Na)/P(Cl) from 4.9 to 2.3 and P(Na)/P(choline) from 3.8 to 2.1, respectively). Transepithelial paracellular basal to apical ⁴⁵Ca fluxes increased approximately twofold when driven by a basal positive Na:NMDG bi-ionic p.d., but with basal 10 mM ZnCl₂, ⁴⁵Ca fluxes decreased approximately twofold. Neither ZO-1 nor occludin distribution was altered after ~2-h exposure to apical 10 mM ZnCl₂. However, claudin-2, though present at the tight junction, increased within the cell. Increased epithelial barrier resistance by Zn²+ is due to modification of the paracellular pathway, most probably by multiple mechanisms.

Sensory and cortical activation of distinct glial cell subtypes in the somatosensory thalamus of young rats

The rodent ventrobasal (VB) thalamus receives sensory inputs from the whiskers and projects to the cortex, from which it receives reciprocal excitatory afferents. Much is known about the properties and functional roles of these glutamatergic inputs to thalamocortical neurons in the VB, but no data are available on how these afferents can affect thalamic glial cells. In this study, we used combined electrophysiological recordings and intracellular calcium ([Ca2+]i) imaging to investigate glial cell responses to synaptic afferent stimulation. VB thalamus glial cells can be divided into two groups based on their [Ca2+]i and electrophysiological responses to sensory and corticothalamic stimulation. One group consists of astrocytes, which stain positively for S100B and preferentially load with SR101, have linear current–voltage relations and low input resistance, show no voltage-dependent [Ca2+]i responses, but express mGluR5-dependent [Ca2+]i transients following stimulation of the sensory and/or corticothalamic excitatory afferent pathways. Cells of the other glial group, by contrast, stain positively for NG2, and are characterized by high input resistance, the presence of voltage-dependent [Ca2+]i elevations and voltage-gated inward currents. There were no synaptically induced [Ca2+]i elevations in these cells under control conditions. These results show that thalamic glial cell responses to synaptic input exhibit different properties to those of thalamocortical neurons. As VB astrocytes can respond to synaptic stimulation and signal to neighbouring neurons, this glial cell organization may have functional implications for the processing of somatosensory information and modulation of behavioural state-dependent thalamocortical network activities.

Single mechano‐gated channels activated by mechanical deformation of acutely isolated cardiac fibroblasts from rats

Mechanosensitive conductances were reported in cardiac fibroblasts, but the properties of single channels mediating their mechanosensitivity remain uncharacterized. The aim of this work was to investigate single mechano-gated channels (MGCs) activated by mechanical deformations of cardiac fibroblasts. Currents through single MGCs and mechanosensitive whole-cell currents were recorded from isolated rat atrial fibroblasts using the cell-attached and whole-cell patch-clamp configurations respectively. Defined mechanical stress was applied via the patch pipette used for the whole-cell recordings. Under resting conditions occasional short openings of two types of single MGCs with conductances of 43 and 87 pS were observed. Both types of channels displayed a linear current-voltage relationship with the reversal potential around 0 mV. Small (1 microm) mechanical deformations affected neither single nor whole-cell mechano-gated currents. Cell compressions (2, 3 and 4 microm) augmented the whole-cell currents and increased the frequency and duration of single channel openings. Cell stretches (2, 3 and 4 microm) inactivated the whole-cell currents and abolished the activity of single MGCs. Gd(3+) (8 microm) blocked the whole-cell currents within 5 min. No single channel activity was observed in the cell-attached mode when Gd(3+) was added to the intrapipette solution. Cytochalasin D and colchicine (100 microm each) completely blocked both the whole-cell and single channel currents. These findings show that rat atrial fibroblasts express two types of MGCs whose activity is governed by cell deformation. We conclude that fibroblasts can sense the direction of applied stress and contribute to mechano-electrical coupling in the heart.

Activation of Polycystic Kidney Disease-2-like 1 (PKD2L1)-PKD1L3 Complex by Acid in Mouse Taste Cells

Five basic tastes (bitter, sweet, umami, salty, and sour) are detected in the four taste areas where taste buds reside. Although molecular mechanisms for detecting bitter, sweet, and umami have been well clarified, those for sour and salty remain poorly understood. Several channels including acid-sensing ion channels have been proposed as candidate sour receptors, but they do not encompass all sour-sensing abilities in vivo. We recently reported a novel candidate for sour sensing, the polycystic kidney disease-2-like 1 (PKD2L1)-PKD1L3 channel complex. This channel is not a traditional ligand-gated channel and is gated open only after removal of an acid stimulus, called an off response. Here we show that off responses upon acid stimulus are clearly observed in native taste cells from circumvallate, but not fungiform papillae, of glutamate decarboxylase 67-green fluorescent protein (GAD67-GFP) knock-in mice, from which Type III taste cells can be visualized, using Ca(2+) imaging and patch clamp methods. Off responses were detected in most cells where PKD2L1 immunoreactivity was observed. Interestingly, the pH threshold for acid-evoked intracellular Ca(2+) increase was around 5.0, a value much higher than that observed in HEK293 cells expressing the PKD2L1-PKD1L3 complex. Thus, PKD2L1-PKD1L3-mediated acid-evoked off responses occurred both in HEK293 cells and in native taste cells, suggesting the involvement of the PKD2L1-PKD1L3 complex in acid sensing in vivo.

Gender‐dependent regulation of expression of the KCNQ1 channel regulatory β‐subunit KCNE3 throughout the estrous cycle in rat distal colonic crypts

The KCNQ1 potassium channel associates with the KCNE3 (‐subunit in the epithelium of the colon and mediates basolateral K+ recycling that is required for apical Cl− secretion. This study tested whether sexual dimorphism exists in the expression and function of the KCNQ1/KCNE3 channel complex in rat distal colonic crypts. The association of KCNE3 with KCNQ1 and the abundance of KCNE3 mRNA and protein were significantly higher in male compared to female crypts. KCNE3 protein expression in the female rat colon varied throughout the estrous cycle; expression was highest during proestrus and declined 4.8‐fold in estrus (P (0.01, n=3) and became undetectable at metaestrus. Whole‐cell K+ currents in male crypts displayed a linear current‐voltage relationship and were 10‐fold more sensitive to chromanol 293B when compared with K+ currents in female crypts which displayed outward rectification. Basolateral membrane K+ currents recorded in colonic epithelia mounted in Ussing chambers also displayed a 10‐fold higher sensitivity to chromanol 293B in male tissues compared to females. This study demonstrates gender and estrous cycle regulation of the expression of KCNE3 and the properties of the KCNQ1 channel activity required for Cl− secretion.Funded by HEA Ireland PRTLI NBIP Cycle 4.

Dependence of ohmic contact resistance on barrier thickness of AlN/GaN HEMT structures

Abstract A multi-faceted study on the reduction of ohmic contact resistance to AlN/GaN-based heterostructures is presented. Minimum contact resistance of 0.5 Ω mm has been achieved by partially etching the AlN barrier layer using a chlorine-based plasma dry-etch prior to ohmic contact metallization. For thin GaN-capped AlN/GaN heterostructures, we find it is necessary to remove the GaN cap in the vicinity of the contact metal in order to obtain a linear current–voltage relationship. We compare our results of the pre-metallization etched contacts to those without an etch as well as to results reported in the literature.

Non-Toxin Gating Modifiers Reveal the Promiscuous Nature of the Voltage Sensor of Kv7.2 and TRPV1 Channels

Voltage-sensitive cation channel architecture consists of two main structural modules, the voltage sensor domain (VSD) and the pore domain. The VSDs are the target of various gating-modifier toxins and their paddle motifs are modular and transferable structures. Here we show that NH17 and NH29, two new Kv7.2 channel blocker and opener, respectively, act as non-toxin gating modifiers. Mutagenesis and modeling data suggest that in Kv7.2, NH29 docks to the external groove formed by the interface of helices S1, S2 and S4 in a way, which stabilizes the interaction between two conserved charged residues in S2, and S4, known to interact electrostatically, in the open state of Kv channels. Reflecting the promiscuous nature of the VSD, NH29 is also a potent blocker of TRPV1 channels, a feature similar to that described for tarantula toxins. TRPV1 channels appear to be weakly voltage-dependent. However, NH29 changes the linear TRPV1 current-voltage relation obtained with capsaicin, to an outwardly rectifying shape. Interestingly, mutations in linker S3-S4 of the TRPV1 VSD are significantly more resistant to the inhibitory effect of NH29. While compound NH17 potently blocks Kv7.2 channels, it sensitizes the TRPV1 current activated by capsaicin. Mutations in linker S3-S4 switch the TRPV1 sensitizing action of NH17 to a potent inhibition. Subtle modifications in the VSD or in the chemical structure of the molecule drastically change the attributes of the gating-modifier, thereby stabilizing the channel in either the closed or the open state. Data indicate that the novel compounds may operate via a voltage-sensor trapping mechanism similar to that suggested for scorpion and sea anemone toxins. Thus, the VSDs of Kv and TRPV1 channels are promiscuous and share some common structural and biophysical features.

Outwardly Rectifying Tonically Active GABAAReceptors in Pyramidal Cells Modulate Neuronal Offset, Not Gain

Hippocampal pyramidal cell excitability is regulated both by fast synaptic inhibition and by tonically active high-affinity extrasynaptic GABA(A) receptors. The impact of tonic inhibition on neuronal gain and offset, and thus on information processing, is unclear. Offset is altered by shunting inhibition, and the gain of a neuronal response to an excitatory input can be modified by changing the level of "background" synaptic noise. Therefore, tonic activation of GABA(A) receptors would be expected to modulate offset and, in addition, to alter gain through a shunting effect on synaptic noise. Here we show that tonically active GABA(A) receptors in CA1 pyramidal cells show marked outward rectification, while the peaks of IPSCs exhibit a linear current-voltage relationship. As a result, tonic GABA(A) receptor-mediated currents have a minimal effect upon subthreshold membrane potential variation due to synaptic noise, but predominantly affect neurons at spiking threshold. Consistent with this, tonic GABA(A) receptor-mediated currents in pyramidal cells exclusively affect offset and not gain. Modulation of tonically active GABA(A) receptors by fluctuations in extracellular GABA concentrations or neuromodulators acting on high-affinity receptors potentially provides a powerful mechanism to alter neuronal offset independently of neuronal gain.

TWIK-1 and TREK-1 Are Potassium Channels Contributing Significantly to Astrocyte Passive Conductance in Rat Hippocampal Slices

Expression of a linear current-voltage (I-V) relationship (passive) K(+) membrane conductance is a hallmark of mature hippocampal astrocytes. However, the molecular identifications of the K(+) channels underlying this passive conductance remain unknown. We provide the following evidence supporting significant contribution of the two-pore domain K(+) channel (K(2P)) isoforms, TWIK-1 and TREK-1, to this conductance. First, both passive astrocytes and the cloned rat TWIK-1 and TREK-1 channels expressed in CHO cells conduct significant amounts of Cs(+) currents, but vary in their relative P(Cs)/P(K) permeability, 0.43, 0.10, and 0.05, respectively. Second, quinine, which potently inhibited TWIK-1 (IC(50) = 85 microm) and TREK-1 (IC(50) = 41 microm) currents, also inhibited astrocytic passive conductance by 58% at a concentration of 200 microm. Third, a moderate sensitivity of passive conductance to low extracellular pH (6.0) supports a combined expression of acid-insensitive TREK-1, and to a lesser extent, acid-sensitive TWIK-1. Fourth, the astrocyte passive conductance showed low sensitivity to extracellular Ba(2+), and extracellular Ba(2+) blocked TWIK-1 channels at an IC(50) of 960 microm and had no effect on TREK-1 channels. Finally, an immunocytochemical study showed colocalization of TWIK-1 and TREK-1 proteins with the astrocytic markers GLAST and GFAP in rat hippocampal stratum radiatum. In contrast, another K(2P) isoform TASK-1 was mainly colocalized with the neuronal marker NeuN in hippocampal pyramidal neurons and was expressed at a much lower level in astrocytes. These results support TWIK-1 and TREK-1 as being the major components of the long-sought K(+) channels underlying the passive conductance of mature hippocampal astrocytes.

The circular bacteriocin, carnocyclin A, forms anion-selective channels in lipid bilayers

Bacterial resistance to conventional antibiotics is a major challenge in controlling infectious diseases and has necessitated the development of novel approaches in antimicrobial therapy. One such approach is the use of antimicrobial peptides, such as the bacterially produced bacteriocins. Carnocyclin A (CclA) is a 60-amino acid circular bacteriocin produced by Carnobacterium maltaromaticum UAL307 that exhibits potent activity against many Gram-positive bacteria. Lipid bilayer and single channel recording techniques were applied to study the molecular mechanisms by which CclA interacts with the lipid membrane and exerts its antimicrobial effects. Here we show that CclA can form ion channels with a conductance of 35 pS in 150 mM NaCl solution. This channel displays a linear current–voltage relationship, is anion-selective, and its activation is strongly voltage-dependent. The formation of ion channels by CclA is driven by the presence of a negative membrane potential and may result in dissipation of membrane potential. Carnocyclin A's unique functional activities as well as its circular structure make it a potential candidate for developing novel antimicrobial drugs.

Preparation and Electrical Properties of the Bi0.9La0.1Fe0.9Mn0.1O3/SrTiO3:Nb p-n Heterojunctions

We report synthesis of single phase highly [100]-axis oriented Bi0.9La0.1Fe0.9Mn0.1O3 (BLFMO) thin films grown by RF sputtering at 650–750°C under Ar/O2 (1:1) gas pressure. The current-voltage characteristics of the p-n heterostructures prepared by growing the BLFMO films on highly conductive 0.7 weight % Nb-doped SrTiO3(100) (STON) have been investigated. The current-voltage dependencies in a case of a forward bias have been modeled taking into account linear current-voltage relationship caused by space charge-limited current at low bias voltages, as well as major contribution of Shottky emission and current limiting effect by series resistance at higher forward bias voltages.