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Ca2+-activated K+-current density is correlated with soma size in rat vestibular-afferent neurons in culture.

Vestibular-afferent neurons (VANs) transmit information about linear and angular accelerations during head movements from vestibular end organs to vestibular nuclei. In situ, these neurons show heterogeneous discharge patterns that may be produced by differences in their intrinsic properties. However, little is known about the ionic currents underlying their different firing patterns. Using the whole cell patch-clamp technique, we analyzed the expression of Ca(2+) and Ca(2+)-activated K(+) currents (I(KCa)) in primary cultured neurons isolated from young rats (p7-p10). We found two overlapping subpopulations of VANs classified according to low-threshold Ca(2+)-current [low-voltage-activated (LVA)] expression; LVA (-) neurons, formed by small cells, and LVA (+) neurons composed of medium to large cells. The I(KCa) in both cell-groups was carried through channels of high (BK), intermediate (IK), and low conductance (SK), besides a resistant channel to classical blockers (IR). BK was expressed preferentially in LVA (+) cells, whereas IR expression was preferentially in LVA (-) cells. No correlation between SK and IK expression with the soma size was found. Current-clamp experiments showed that BK participates in the adaptation of discharge and in the duration of the action potential, whereas SK and IK did not show a significant contribution to electrical discharge of cultured VANs. However, because of the low number of VANs in culture with repetitive firing it is difficult to interpret our results in terms of discharge patterns. Our results demonstrate that vestibular-afferent neurons possess different Ca(2+)-activated K(+) (K(Ca)) channels and that their expression, heterogeneous among the cells, would contribute to explain some of the differences in the electrical-firing properties of these neurons.

4-Aminopyridine↗

Estrogen and inflammation increase the excitability of rat temporomandibular joint afferent neurons.

Several painful conditions, including temporomandibular disorders (TMD), are more prevalent and more severe in women than in men. Although the physiological basis for this sex difference remains to be determined, it is likely that estrogen is an underlying factor. The present study was performed to test the hypotheses that estrogen increases the excitability of rat temporomandibular joint (TMJ) afferents and exacerbates the inflammation-induced sensitization of these sensory neurons. Retrogradely labeled TMJ neurons from ovariectomized rats and ovariectomized rats receiving chronic estrogen replacement were studied using whole cell patch-clamp techniques three days after injecting the TMJ with either saline or Complete Freund's Adjuvant to induce inflammation. Excitability was assessed with depolarizing current injection to determine action potential threshold, rheobase, and the response to suprathreshold stimuli. Spontaneous activity was also assessed. Both inflammation and estrogen increased the excitability of TMJ neurons as reflected by decreases in action potential threshold and rheobase and increases in the incidence of spontaneous activity. The effects were additive with neurons from rats receiving both estrogen and inflammation being the most excitable. The increases in excitability were associated with changes in passive properties and action potential waveform, suggesting that estrogen and inflammation affect the expression and/or properties of ion channels in TMJ neurons. Importantly, the influence of estrogen on both baseline and inflammation-induced changes in TMJ neuronal excitability may help explain the profound sex difference observed in TMD as well as suggest a novel target for the treatment of this pain condition.

Action Potentials↗

Activation of GIRK channels by muscarinic receptors and group II metabotropic glutamate receptors suppresses Golgi cell activity in the cochlear nucleus of mice.

Granule cells and parallel fiber circuits in the dorsal cochlear nucleus (DCN) play a role in integration of multimodal sensory with auditory inputs. The activity of granule cells is regulated through inhibitory connections made by Golgi cells. Golgi cells in turn probably receive parallel fiber inputs and regulate activity of the DCN. We have investigated the electrophysiological properties of Golgi cells using the whole cell patch-clamp method in slices made from transgenic mice that express green fluorescent protein driven by the promotor of metabotropic glutamate receptor subtype 2. Stimulation of auditory nerve fibers (ANFs) and of parallel fibers evoked glutamatergic excitatory postsynaptic currents (EPSC) through AMPA receptors. The strengths and latencies of these inputs differed, however. ANF stimulation evoked EPSCs after 4.7 +/- 0.4 ms, whereas parallel fiber stimulation evoked EPSCs after 1.4 +/- 0.2 ms that were on average 2.5 times as large. The multiple peaks and prolonged activity suggest the presence of polysynaptic connections between ANFs and Golgi cells. Agonists for group II metabotropic glutamate receptors (mGluRs) and for muscarinic receptors induced membrane hyperpolarization and suppressed the firing of Golgi cells by activating G-protein-coupled inward rectifier K(+) (GIRK) channels. These results strongly suggest that Golgi cells were regulated through the combined activities of glutamatergic and cholinergic synapses, which presumably regulated the temporal firing patterns of granule cells and through them the activity of principal cells of the DCN.

Action Potentials↗

Do urea/ammonium lactate combinations achieve better skin protection and hydration than either component alone?

Twenty subjects with healthy skin were treated with the following formulations for two weeks: drug-free W/O vehicle, 5% ammonium lactate (CAS 52003-58-4) in W/O vehicle, 5% urea (CAS 57-13-6) in W/O vehicle, 3% ammonium lactate + 3% urea in W/O vehicle, 5% ammonium lactate + 5% urea in W/O vehicle. These formulations were applied in randomized order to 6 test areas on the forearms; one area was left untreated. Repetitive washings were additionally performed in the second treatment week. TEWL, stratum corneum water content (corneometry-determined electrical capacitance), and cutaneous blood flow (laser Doppler) were measured at baseline, day 7, and day 14. 5% urea and both ammonium lactate/urea combinations produced significant stratum corneum hydration and improved stratum corneum barrier function; there were no significant differences between these three treatments.

Adult↗

Na+-Ca2+ exchange activity is localized in the T-tubules of rat ventricular myocytes.

Detubulation of rat ventricular myocytes has been used to investigate the role of the t-tubules in Ca2+ cycling during excitation-contraction coupling in rat ventricular myocytes. Ca2+ was monitored using fluo-3 and confocal microscopy. In control myocytes, electrical stimulation caused a spatially uniform increase in intracellular [Ca2+] across the cell width. After detubulation, [Ca2+] rose initially at the cell periphery and then propagated into the center of the cell. Application of caffeine to control myocytes resulted in a rapid and uniform increase of intracellular [Ca2+]; the distribution and amplitude of this increase was the same in detubulated myocytes, although its decline was slower. On application of caffeine to control cells, there was a large, rapid, and transient rise in extracellular [Ca2+] as Ca2+ was extruded from the cell; this rise was significantly smaller in detubulated cells, and the remaining increase was blocked by the sarcolemmal Ca2+ ATPase inhibitor carboxyeosin. The treatment used to produce detubulation had no significant effect on Ca2+ efflux in atrial cells, which lack t-tubules. Detubulation of ventricular myocytes also resulted in loss of Na+-Ca2+ exchange current, although the density of the fast Na+ current was unaltered. It is concluded that Na+-Ca2+ exchange function, and hence Ca2+ efflux by this mechanism, is concentrated in the t-tubules, and that the concentration of Ca2+ flux pathways in the t-tubules is important in producing a uniform increase in intracellular Ca2+ on stimulation.

Adenosine Triphosphatases↗

Modulation of an inactivating human cardiac K+ channel by protein kinase C.

The transient outward current (ITO) is an important repolarizing component of the cardiac action potential. In native cardiac myocytes, ITO is modulated after activation of protein kinase C, although the molecular nature of this effect is not well understood. A channel recently cloned from human ventricular myocardium (Kv1.4, HK1) produces a rapidly inactivating K+ current, which has phenotypic similarities to the 4-aminopyridine-sensitive component of ITO. Therefore, we examined whether this recombinant channel was also modulated by protein kinase C activation by investigating the effects of the diacylglycerol analogue phorbol 12-myristate 13-acetate (PMA) on Kv1.4 K+ current expressed in Xenopus oocytes. At a concentration of 10 nmol/L, PMA caused a biphasic response with an initial increase (14 +/- 4%, mean +/- SEM) in current, which peaked in 14 minutes. This was followed by a significant reduction (40 +/- 11%) in the current within 30 minutes. There was no significant change in cell membrane electrical capacitance with 10 nmol/L PMA (1 +/- 1% decline in 30 minutes), demonstrating that loss of cell membrane surface area did not explain the reduction in K+ current, although cell capacitance did decrease when using a higher concentration of PMA (81 nmol/L). The inactive stereoisomer, 4 alpha-PMA, had no effect on Kv1.4 current, whereas preincubation with the protein kinase inhibitor staurosporine or protein kinase C-selective chelerythrine prevented the effects of PMA. When purified from a stably transfected mammalian cell line by using immunoprecipitation, the channel protein was readily phosphorylated in vitro by purified protein kinase C.(ABSTRACT TRUNCATED AT 250 WORDS)

Action Potentials↗

Mechanism of burn injury during magnetic resonance imaging (MRI)--simple loops can induce heat injury.

To determine the mechanism of burn injury associated with magnetic resonance imaging (MRI), the induced current in the loops of a conductive lead was measured. Seven types of loops with effective areas within the range from 100 to 12 000 cm2 were made and then each loop was placed in the bore of a whole-body MR system at 0.5 T and MRI was carried out. During radio frequency (RF) irradiation, an induced voltage was observed in loops that were placed with their axes parallel to the linearly polarized transmitting RF field. The voltage had a sincfunction characteristic and was within the range 55-235 V at the RF pulse sequence for usual MR imaging. When the axis was vertical, negligible current was observed. A resistor inserted into the circuit of a 30 x 40 cm loop sparked and burned out. Simple loops of conductive material may result in the induction of a large and potentially hazardous voltage in the imaging system. Self-resonance of such a loop may add greatly to hazards by increasing the effective coupling to the RF transmitter. Also, impedance matching of the inserted resistor with the impedance of the loop increases the dissipated power at the resistor.

Burns, Electric↗

Phase evolution, characterization, and impedance spectroscopic analysis of nanocrystalline SrBi2Nb2O9 in glassy Li2B4O7 matrix.

Transparent glass composites in the system (100 - x) Li2B4O7 - x SrBi2Nb2O9 (where x = 5, 10, 15, 20, 25, and 30, in molar ratio) were fabricated by conventional melt-quenching technique. The amorphous nature of the as-quenched glass composites was confirmed via X-ray powder diffraction (XRD) studies. Differential thermal analyses (DTA) established the glassy nature of the as-quenched samples. Glass nanocomposites (GNC's) with high-optical transparency were obtained by controlled heat-treatment of the glass composites at 750 K/6 h. Perovskite SrBi2Nb2O9 (SBN) phase formation through an intermediate fluorite phase was confirmed by XRD and transmission electron microscopy (TEM). The dielectric constant (epsilon r) in the frequency range from 100 Hz to 40 MHz at room temperature increases whereas dielectric loss (D) decreases with increase in SBN content in Li2B4O7 glass matrix. Impedance spectroscopy employed to rationalize the electrical behavior of the as-quenched glasses and glass nanocomposites suggests the coexistence of electronic and ionic conduction in these materials. The optical transmission and band-gap energy of these composites were found to be crystallite size dependent.

Bismuth↗

Dielectric properties of KDP filled porous alumina nanocomposite thin films.

A new concept of a composite dielectric thin film fabrication is presented. The fabrication process consists of two stages. The first stage is anodizing a thin aluminum film to produce a porous alumina film that contains an array of nanometer sized parallel pores. The second stage is filling the pores with a saturated KDP (KH2PO4) liquid solution due to capillary forces. After drying KDP nanocrystals are formed inside the pores. This process results in a formation of a composite dielectric thin film composed of the alumina pores walls as one dielectric material and the KDP nanocrystals inside the pores as another dielectric material. The dielectric permittivity of this composite film is higher than that of the porous alumina film at all applied frequencies. The dielectric enhancement is more pronounced at low frequencies due to an interface polarization mechanism. This fabrication process enables controlling the size, composition, and microstructure of the composite dielectric film constituents and thus changing its dielectric properties over a wide range of values.

Aluminum Oxide↗

NEGF-HF method in molecular junction property calculations.

Electron-electron interaction is an essential issue in predicting the properties of a molecular conduction junction. An accurate treatment requires taking proper account of the potential distribution across the junction as well as for the change in electronic structure under the external voltage drop. Another important point is the necessity to treat molecules as open quantum systems. In this paper we address these issues within the framework of the non-equilibrium Green's function formalism at the Hartree-Fock level (NEGF-HF), which permits a self-consistent treatment of the problem. We apply the method to study the experimentally observed asymmetry in the I-V curves with respect to polarity of voltage bias in Hg-Au junctions containing bilayers of alkanethiols of various chain lengths. The origin of the effect is suggested to be the asymmetric behavior of the character of the highest occupied molecular orbital (HOMO) of the junction at opposite biases, which leads to different effective barriers for electron transfer across the junction at opposite signs of the voltage drop. The calculated potential profile shows the capacitor-like nature for the junction with the weak link.

Algorithms↗

A simple theoretical model to study the voltage dependence of the electronic structure of phenyl ethylene oligomers.

Although very few measurements have appeared in the open literature and there seems to be a controversy about the existence of the NDR phenomenon in molecules, the prospects of building such systems have attracted significant attention. In the work reported in this paper we used a model based on DFT calculations of the electronic structure of the 2'-amino-4,4'-di(ethynylphenyl)-5'-nitro-1-benzenethiolate molecule (previously reported to exhibit NDR behavior) in a capacitor-like electric field that mimics the potential spatial profile of the junction. Our results suggest that in these systems, there seems to be a correlation between a substantial charge density rearrangement of the neutral bridge at a threshold voltage and the NDR behavior observed in previous experiments. Our results highlight the importance of inclusion of the field in the study of electrified interfaces. We applied this model to a fluorine-substituted conjugated diethynylphenyl molecule and found that these calculations predict similar behavior. Results based on extended system calculations, including electrode-molecule interactions, confirm the validity of the model based on the isolated molecule and suggest the use of these simple models to rationally design molecular devices with similar switching characteristics.

Algorithms↗

Scalability simulations for nanomemory systems integrated on the molecular scale.

Simulations were performed to assess the prospective performance of a 16 Kbit nanowire-based electronic nanomemory system. Commercial off-the-shelf microcomputer system modeling software was applied to evaluate the operation of an ultra-dense storage array. This array consists of demonstrated experimental non-volatile nanowire diode switches, plus encoder-decoder structures consisting of demonstrated experimental nanowire-based nanotransistors, with nanowire interconnects among all the switching devices. The results of these simulations suggest that a nanomemory of this type can be operated successfully at a density of 10(11) bits/cm(2). Furthermore, modest device alterations and system design alternatives are suggested that might improve the performance and the scalability of the nanomemory array. These simulations represent early steps toward the development of a simulation-based methodology to guide nanoelectronic system design in a manner analogous to the way such methodologies are used to guide microelectronic system design in the silicon industry.

Computer Simulation↗

Effect of prenatal steroids on skin surface hydrophobicity in the premature rat.

The skin surface of the newborn rat at term is highly hydrophobic. This surface hydrophobicity plays a putative role in the transition from an aqueous to a gaseous environment at birth and is dependent on the presence of an intact periderm. Glucocorticoids given to pregnant dams, during late gestation, will accelerate formation of the stratum corneum and reduce transepidermal water loss in prematurely delivered pups. We tested the related hypotheses that surface hydrophobicity and maturation of the periderm are developmentally accelerated by prenatal exposure to steroids. Thirty pregnant Sprague-Dawley rats received either normal saline or 0.5 mg/kg betamethasone on d 17 of gestation. After cesarean delivery on d 18, 19, and 20, dorsal skin surface hydrophobicity was quantified by direct surface electrical capacitance (SEC) measurement. Initial skin surface hydration at birth was significantly lower in steroid-treated pups than in control pups at gestational ages 19 and 20 d (3060 +/- 1379 versus 4441 +/- 153 pF and 646 +/- 295 versus 1493 +/- 1019 pF, respectively, p < 0.001, mean +/- SD). Likewise, after desorption of amniotic fluid, baseline skin hydration was significantly lower in steroid-treated pups than in control pups at gestational ages 19 and 20 d (1862 +/- 1560 pF versus 4278 +/- 97 pF and 60 +/- 56 pF versus 128 +/- 264 pF, p < 0.001). Scanning and transmission electron microscopy showed morphologic maturation of the periderm after steroid treatment. These results demonstrate accelerated development of both functional and structural correlates of skin surface hydrophobicity in the premature rat after prenatal exposure to steroids.

Animals↗

Neuronal basis of the slow (<1 Hz) oscillation in neurons of the nucleus reticularis thalami in vitro.

During deep sleep and anesthesia, the EEG of humans and animals exhibits a distinctive slow (<1 Hz) rhythm. In inhibitory neurons of the nucleus reticularis thalami (NRT), this rhythm is reflected as a slow (<1 Hz) oscillation of the membrane potential comprising stereotypical, recurring "up" and "down" states. Here we show that reducing the leak current through the activation of group I metabotropic glutamate receptors (mGluRs) with either trans-ACPD [(+/-)-1-aminocyclopentane-trans-1,3-dicarboxylic acid] (50-100 microM) or DHPG [(S)-3,5-dihydroxyphenylglycine] (100 microM) instates an intrinsic slow oscillation in NRT neurons in vitro that is qualitatively equivalent to that observed in vivo. A slow oscillation could also be evoked by synaptically activating mGluRs on NRT neurons via the tetanic stimulation of corticothalamic fibers. Through a combination of experiments and computational modeling we show that the up state of the slow oscillation is predominantly generated by the "window" component of the T-type Ca2+ current, with an additional supportive role for a Ca2+-activated nonselective cation current. The slow oscillation is also fundamentally reliant on an Ih current and is extensively shaped by both Ca2+- and Na+-activated K+ currents. In combination with previous work in thalamocortical neurons, this study suggests that the thalamus plays an important and active role in shaping the slow (<1 Hz) rhythm during deep sleep.

Action Potentials↗

N-terminal-mediated homomultimerization of prestin, the outer hair cell motor protein.

The outer hair cell lateral membrane motor, prestin, drives the cell's mechanical response that underpins mammalian cochlear amplification. Little is known about the protein's structure-function relations. Here we provide evidence that prestin is a 10-transmembrane domain protein whose membrane topology differs from that of previous models. We also present evidence that both intracellular termini of prestin are required for normal voltage sensing, with short truncations of either terminal resulting in absent or modified activity despite quantitative findings of normal membrane targeting. Finally, we show with fluorescence resonance energy transfer that prestin-prestin interactions are dependent on an intact N-terminus, suggesting that this terminus is important for homo-oligomerization of prestin. These domains, which we have perturbed, likely contribute to allosteric modulation of prestin via interactions among prestin molecules or possibly between prestin and other proteins, as well.

Allosteric Site↗

Gap junctions and connexon hemichannels in human embryonic stem cells.

Intercellular communication via gap junctions is thought to play an important role in embryonic cell survival and differentiation. Classical studies demonstrated both dye and electrical coupling of cells in the inner cell mass of mouse embryos, as well as the development of restrictions against coupling between cells of the inner cell mass and surrounding trophectoderm. Here we demonstrate extensive gap junctional communication between human embryonic stem (ES) cells, the pluripotent cells isolated from the inner cell mass of preimplantation blastocysts. Human ES cells maintained in vitro expressed RNA for 18 of the 20 known connexins; only connexin 40.1 (Cx40.1) and Cx50 were not detected by reverse transcription-polymerase chain reaction. Cx40, Cx43, and Cx45 were visualized by immunofluorescence at points of contact between adjacent cells. Electron microscopy confirmed that neighboring cells formed zones of tight membrane apposition characteristic of gap junctions. Fluorescent dye injections demonstrated extensive coupling within human ES cell colonies growing on mouse embryonic fibroblast (MEF) feeder cells, whereas dye coupling between human ES cells and adjacent MEFs was extremely rare. Physiological recordings demonstrated electrical and dye coupling between human ES cells in feeder-free monolayers and between isolated human ES cell pairs. Octanol, 18-alpha-glycyrrhetinic acid, and arylaminobenzoates inhibited transjunctional currents. Dye uptake studies on human ES cell monolayers and recordings from solitary human ES cells gave evidence for the surface expression of connexon hemichannels. Human ES cells provide a unique system for the study of human connexin proteins and their potential functions in cellular differentiation and the maintenance of pluripotency.

Animals↗

Application of electrocapacitography to investigation of cephalic and cerebral circulation in the rabbit.

Electrocapacitography devised for medical purposes by Hatakeyama was applied as a noninvasive volumetric method to investigate circulation in the rabbit head and brain. The effective electric capacitance was measured by means of a highly sensitive device. An electrocapacitogram (ECPG) of the head, considered to reflect the volume of intracranial as well as extracranial blood vessels, varied rhythmically with cardiac beat and respiratory movement. In general, the pulsatile variations of ECPG in synchronization with cardiac beat (ECPG pulse) bore a resemblance to arterial pulse. Occlusion of the common carotid artery caused the ECPG level corresponding to mean vascular volume to fall markedly and in most cases the amplitude of ECPG pulse was diminished. By rapid infusion of 5 ml of the blood into intracranial blood vessels via the internal carotid artery or into extracranial blood vessels via the external carotid artery, ECPG level was raised transitorily, the effect of extracranial infusion being larger than that of intracranial infusion. Stimulation of the central cut end of the cervical sympathetic nerve reduced ECPG level considerably. This response was greatly diminished by carotid occlusion and sometimes a temporary rise of ECPG was observed at the beginning of the stimulation. In several cases the effect was reversed completely by carotid occlusion and an increase of ECPG during the stimulation was observed. These findings suggest the existence of a sympathetic vasoconstrictor in the head as well as a sympathetic vasodilator of the brain, although in confirmation of this review further investigation is required. ECPG of the head is demonstrated to be a useful method for investigating circulation in the head including brain.

Animals↗