Effects of different ions on resting polarization and on the mass receptor potential of carotid body chemosensors

The carotid body and its own nerve were removed from cats anesthetized with sodium pentobarbital and placed in an air gap system; the carotid body was bathed in modified Locke's solution equilibrated with 50% O₂ in N₂, pH 7.43 at 35°C. The sensory discharges, changes in “resting” receptor polarization and the mass receptor potential evoked by ACh or NaCN were recorded with nonpolarizable electrodes placed across the gap. Receptor potentials and sensory discharges evoked by ACh showed an appreciable increase in amplitude and frequency when the preparation was bathed in eserinized Locke. Eserine did not change appreciably the responses evoked by NaCN. Excessive depolarization elicited by either ACh or NaCN was accompanied by sensory discharge block. Removal of K⁺ ions from the bathing solution induced receptor hyperpolarization and an increase in the amplitude of the evoked receptor potentials. An increase of K⁺ concentration had the opposite effect. Reduction of Na⁺ or NaCl to one half, or total removal of this salt, induced an initial reduction and later disappearance of the sensory discharges, some receptor hyperpolarization and a reduction in the amplitude of the evoked receptor potentials. Reduction or removal of Ca⁺⁺ produced receptor depolarization, a marked depression of the evoked receptor potentials, an increase in the frequency of the sensory discharges and a reduction in the amplitude of the nerve action potentials. High Ca⁺⁺ or Mg⁺⁺ had little or no effect on action potential amplitude or resting polarization, but decreased sensory discharge frequency and the evoked receptor potentials. Total or partial replacement of Ca⁺⁺ with Mg⁺⁺ induced complex effects: (1) receptor depolarization which occurred in low Ca⁺⁺, was prevented by addition of Mg⁺⁺ ions; (2) the amplitude of the evoked receptor potentials was depressed; (3) the nerve discharge frequency was reduced as it was in high Mg⁺⁺ solutions; and (4) the amplitude of the nerve action potentials was reduced as it was in low Ca⁺⁺ solutions. Temperature had a marked effect on the chemoreceptors since a t high temperatures the receptors were depolarized and the discharge frequency increased. The baseline discharge and responses evoked by ACh or NaCN were depressed at low temperatures. The results are discussed in terms of possible receptor mechanisms influenced by the different ions.     

Specificity of the K-channels in labyrinthine organs: effects of Rb+ and Cs+ on the conversion process in frog semicircular canals

Summary The specificity of the K-channels which are present in the ciliated membrane of ampullar receptors has been investigated by replacing K⁺ in the fluids bathing the canal with Rb⁺ and Cs⁺. Results show that, unlike Cs⁺, Rb⁺ is able to substitute K⁺ in maintaining the receptor function. These findings favour the hypothesis that the transduction channels which allow the receptor current to flow across sensory cell bodies are specific K-channels. The effects of Rb⁺ and Cs⁺ on primary sensory neuron endings were also studied.Abbreviations Adc slow ampullar potentials - Ndc slow nerve potentials     

Antennal thermo- and hygrosensitive sensilla in Antheraea pernyi (Lepidoptera,Saturniidae): ultrastructure and immunohistochemical localization of Na+,K+-ATPase

Summary In an attempt to identify and localize the components of voltage sources involved in sensory transduction in insect sensilla, the thermo-/hygrosensitive sensilla of the moth Antheraea pernyi were probed with a polyclonal antiserum against Na⁺,K⁺-ATPase in cryofixed and freeze-substituted preparations. The antiserum recognized epitopes on the cytoplasmic membranes of the dendritic inner segments and somata of the sensory cells and also on the cytoplasmic membranes of glial cells surrounding the initial axon segments. The findings support the current concept that ion pumps in the cytoplasmic membranes of the dendritic inner segments and somata of the sensory cells contribute to the maintenance of the resting potential of the sensory cells and to the driving forces generating the receptor currents in response to stimulation of the sensillum. Morphological features and immunohistochemical characteristics of the region of the initial axon segment are also discussed with respect to the initiation of action potentials in these sensilla.     

On the kinetics of potential,electromotance, and chemical change in the excitable system of nerve

The kinetics of interaction between potential, chemical equilibrium, and electromotance in the excitable system of nerve are analyzed. The theoretical system has the following properties: It gives rise to two electromotances each of which depends directly on a chemical equilibrium. The equilibria are determined by the potential across the system. After a sudden potential shift the equilibria reach their new value with an exponential time course, the time constant of which is determined by the rate constants of the two reactions. The rate constants are different due to different activation energies. The two electromotances give rise to potentials of opposite sign. The total potential produced by the system is equal to the sum of the two potentials. The two equilibria are thus determined by any externally applied potential as well as by the sum of the internally produced potentials. The dependence of the equilibria on the potential is calculated from first principles. The equations which describe this system are solved by an analogue computer, which gives instantaneous solutions of the total internal potential as a function of time and any voltage applied from an external source. Comparison between recorded and computed action potentials shows excellent agreement under all experimental conditions. The electromotances might originate from a Ca⁺⁺—Na⁺—K⁺ exchange at fixed negative sites in the Schwann cell.     

Altered mechanoreceptor response in Drosophila bang-sensitive mutants

Bang-sensitive mutants of Drosophila melano gaster (bas ¹, bss^MW1, eas², tko^25t) display seizure followed by paralysis when subjected to mechanical shock. However, no physiological or biochemical defect has been found to be common to all of these mutants. In order to observe the effects of bang-sensitive mutations upon an identified neuron, and to study the nature of mechanically induced paralysis, we examined the response of a mechanosensory neuron in these mutants. In each single mutant and the double mutant bas ¹ bss^MW1, the frequency of action potentials in response to a bristle displacement was reduced. This is the first demonstration of a physiological defect common to several of the bang-sensitive mutations. Adaptation of spike frequency, cumulative adaptation to repeated stimulation (fatigue) and the time course of recovery from adaptation were also examined. Recovery from adaptation to a conditioning stimulus was examined in two mutants (bas ¹ and bss ^MW1), and initial recovery from adaptation was greater in both mutants. Quantification of receptor potentials was complicated by variability inherent in extracellular recording conditions, but examination of the waveform and range of amplitudes did not indicate clear mutant defects. Therefore the differences observed in the spike response may be due to an alteration of the transfer from receptor potentials to action potential production. DNA sequence analysis of tko and eas has indicated that they encode apparently unrelated biochemical products. Our results suggest that these biochemical lesions lead to a common physiological defect in mechanoreceptors. Although this defect does not provide a straightforward explanation for bang sensitivity, the altered cellular process may lead to bang sensitivity through its action in different parts of the nervous system.Abbreviations APA anterior post-alar - ANP anterior notopleural - bas bang-sensitive - bss bang-senseless - eas easily-shocked tko technical knockout     

Allosteric Effects of Permeating Cations on Gating Currents during K+ Channel Deactivation

K⁺ channel gating currents are usually measured in the absence of permeating ions, when a common feature of channel closing is a rising phase of off-gating current and slow subsequent decay. Current models of gating invoke a concerted rearrangement of subunits just before the open state to explain this very slow charge return from opening potentials. We have measured gating currents from the voltage-gated K⁺ channel, Kv1.5, highly overexpressed in human embryonic kidney cells. In the presence of permeating K⁺ or Cs⁺, we show, by comparison with data obtained in the absence of permeant ions, that there is a rapid return of charge after depolarizations. Measurement of off-gating currents on repolarization before and after K⁺ dialysis from cells allowed a comparison of off-gating current amplitudes and time course in the same cells. Parallel experiments utilizing the low permeability of Cs⁺ through Kv1.5 revealed similar rapid charge return during measurements of off-gating currents at E_Cs. Such effects could not be reproduced in a nonconducting mutant (W472F) of Kv1.5, in which, by definition, ion permeation was macroscopically absent. This preservation of a fast kinetic structure of off-gating currents on return from potentials at which channels open suggests an allosteric modulation by permeant cations. This may arise from a direct action on a slow step late in the activation pathway, or via a retardation in the rate of C-type inactivation. The activation energy barrier for K⁺ channel closing is reduced, which may be important during repetitive action potential spiking where ion channels characteristically undergo continuous cyclical activation and deactivation.     

Distribution of ion channels on taste cells and its relationship to chemosensory transduction

Summary The presence and regional localization of voltagegated ion channels on taste cells inNecturus maculosus were studied. Lingual epithelium was dissected from the animal and placed in a modified Ussing chamber such that individual taste cells could be impaled with intracellular microelectrodes and the chemical environment of the apical and basolateral membranes of cells could be strictly controlled. That is, solutions bathing the the mucosal and serosal surfaces of the epithelium could be exchanged independently and the effects of pharmacological agents could be tested selectively on the apical or basolateral membranes of taste cells. In the presence of amphibian physiological saline, action potentials were elicited by passing brief depolarizing current pulses through the recording electrode. Action potentials provided a convenient assay of voltage-gated ion channels. As in other excitable tissues, blocking current through Na⁺, K⁺, or Ca²⁺ channels had predictable and consistent effects on the shape and magnitude of the action potential. A series of experiments was conducted in which the shape and duration of regenerative action potentials were monitored when the ionic composition was altered and/or pharmacological blocking agents were added to the mucosal or to the serosal chamber. We have found the following: (1) voltage-gated K⁺ channels (delayed rectifier) are found predominately, if not exclusively, on the chemoreceptive apical membrane; (ii) voltage-gated Na⁺ and Ca²⁺ channels are found on the apical (chemoreceptive) and basolateral (synaptic) membrane; (iii) there is a K⁺ leak channel on the basolateral membrane which appears to vary seasonally in its sensitivity to TEA. The nonuniform distribution of voltage-gated K⁺ channels and their predominance on the apical membrane may be important in taste transduction: alterations in apical K⁺ conductance may underlie receptor potentials ellicted by rapid stimuli.     

TTX sensitive plateau potentials in the crayfish slowly adapting stretch receptor neuron

Summary Electrophysiological experiments showed that a tetrodotoxin (TTX) sensitive slowly inactivating Na⁺ current contributed to the excitability of the sensory neuron (SN1) that innervates the slow receptor muscle in the abdominal muscle receptor (MR1) of crayfish, Procambarus clarkii. Following either tetraethylammonium (TEA) blockage of the K⁺ delayed rectifier currents or exposure to high temperature, a depolarizing plateau potential was evoked by the slow Na⁺ current. Ca⁺⁺ substitution by other divalent cations had no effect on the plateau potential, demonstrating that Ca⁺⁺ is not involved in plateau potential genesis. Simultaneous intrasomatic and extraaxonic recordings coupled with 4-aminopyridine (4-AP) exposure indicated that the slowly inactivating Na⁺ current is primarily somatic, and does not contribute significantly to spiking.Abbreviations 4-AP 4-aminopyridine - HAP hyperpolarizing after-potential - MR1 slowly adapting muscle receptor organ - SR1 sensory neuron of MR1 - TEA tetraethylammonium - TTX tetrodotoxin     

Heterogeneity of Photosystem I reaction centers in barley leaves as related to the donation from stromal reductants

The light-response curves of P700 oxidation and time-resolved kinetics of P700⁺ dark re-reduction were studied in barley leaves using absorbance changes at 820 nm. Leaves were exposed to 45 °C and treated with either diuron or diuron plus methyl viologen (MV) to prevent linear electron flow from PS II to PSI and ferredoxin-dependent cyclic electron flow around PSI. Under those conditions, P700⁺ could accept electrons solely from soluble stromal reductants. P700 was oxidized under weak far-red light in leaves treated with diuron plus MV, while identical illumination was nearly ineffective in diuron-treated leaves in the absence of MV. When heat-exposed leaves were briefly illuminated with strong far-red light, which completely oxidized P700, the kinetics of P700⁺ dark reduction was fitted by a single exponential term with half-time of about 40 ms. However, two first-order kinetic components of electron flow to P700⁺ (fast and slow) were found after prolonged leaf irradiation. The light-induced modulation of the kinetics of P700⁺ dark reduction was reversed following dark adaptation. The fast component (half time of 80–90 ms) was 1.5 larger than the slow one (half time of about 1 s). No kinetic competition occurred between two pathways of electron donation to P700⁺ from stromal reductants. This suggests the presence of two different populations of PSI. This revised version was published online in June 2006 with corrections to the Cover Date.     

Computer simulation of the motoneuron pool–muscle complex. I. Input system and motoneuron pool

 The aim of the present study was to simulate the input system and the motoneuron (MN) pool of the MN pool–muscle complex (MNPMC). Input fibers, which can originate from command centers in the central nervous system or from sensory organs, activate the MN pool. They generate sequences of action potentials, the frequency of which is proportional to a time-dependent activation factor (which is an input to the model). Different connection patterns between the input fibers and motor units (MUs) are allowed. For simplicity and since no precise experimental data are available, 70 input fibers and 4 boutons per fiber and MN are simulated (this corresponds approximately to the monosynaptic group-Ia input of the cat medial gastrocnemius muscle). Each bouton generates the same conductance change in the postsynaptic membrane. The MNs are modeled with a single compartment and a homogenous membrane. According to experimental data, the membrane leakage conductance and capacitance are MU dependent. Since the precise relation is unknown: (a) the computed relation between MU contraction force and the MN leakage conductance was taken from a steady-state MNPMC model, and (b) the capacitance was assumed to be proportional to the leakage conductance. The MN membrane includes time- and voltage-dependent ionic channels (fast and slow K⁺ and low- and high-threshold Ca²⁺ channels). The density and time constant of the slow K⁺ channels and the density of the Ca²⁺ channels were fitted to approximate afterhyperpolarization characteristics and frequency-injected current relations of type-identified cat MNs. If the membrane reaches a voltage threshold the MNs generate action potentials, which were simulated by voltage pulses. The activation of the MN pool of the human first dorsal interosseus muscle was simulated with injected and synaptic currents in order to illustrate the size principle, synaptic noise, and other features of muscle activation. It is concluded that the present model reproduces the main properties of the input–output relations of different MN types within a muscle. Together with the simulation of the muscle force and the surface EMG, which will be published in subsequent papers, it will be a powerful tool for reproducing experiments on the motor system and investigating functional mechanisms of motor control. Received: 17 April 2001 / Accepted in revised form: 6 November 2001     

Involvement of apamin-sensitive SK channels in spike frequency adaptation of neurons in nucleus tractus solitarii of the rat

We delineated the role of Ca²⁺-activated K⁺ channels in the phenomenon of spike frequency adaptation (SFA) exhibited by neurons in the caudal region of nucleus tractus solitarius (cNTS) using intracellular recording coupled with the current-clamp technique in rat brain slices. Intracellular injection of a constant depolarizing current evoked a train of action potentials whose discharge frequency declined rapidly to a lower steady-state level of irregular discharges. This manifested phenomenon of SFA was found to be related to extracellular Ca²⁺. Low Ca²⁺ (0.25 mM) or Cd²⁺ (0.5 mM) in the perfusing medium resulted in a significant increase in the adaptation time constant (_adap) and an appreciable reduction in the percentage adaptation of spike frequency (F_adap). In addition, the evoked discharges were converted from an irregular to a regular pattern, accompanied by a profound increase in mean firing rate. Intriguingly, similar alterations in _adap, F_adap, discharge pattern and discharge rate were elicited by apamin (1 µM), a selective blocker for small-conductance Ca²⁺-activated K⁺ (SK) channels. On the other hand, charybdotoxin (0.1 µM), a selective blocker for large-conductance Ca²⁺-activated K⁺ channels, was ineffective. Our results suggest that SK channels of cNTS neurons may subserve the generation of both SFA and irregular discharge patterns displayed by action potentials evoked with a prolonged depolarizing current.     

A three-state model for inactivation of sodium permeability

The inactivation of Na⁺ permeability in single myelinated motor nerve fibres of Rana esculenta was investigated under voltage and current clamp conditions at 20°C in Ringer's solution and under blocked K⁺ currents. Development of inactivation and its recovery was described by two potential-dependent time constants: The smaller time constant followed the usual bell-shaped function of membrane potential, whereas the larger one was monotone-increasing with more negative potentials. Several three-state models for inactivation were investigated. The experiments could best be approximated by a model with two open and one closed state for inactivation following: open ? closed ? open. Rate constants were determined for all transitions shown from the voltage clamp experiments. The action potentials computed by means of the proposed model were in good agreement with those measured, both in Ringer's solution and under blocked K⁺ current conditions.     

The depolarizing mechanoreceptor potential and Ca/Mg receptor-current of the marine ciliate Euplotes vannus

The anterior depolarizing mechanoreceptor potential and the correlated receptor currents have been studied in the marine ciliate Euplotes vannus. Mechanical stimuli that mimicked cell-cell collisions depolarized the resting potential of about — 25 mV to maximally — 5 mV, with a speed of 1.2 mV/ms, a delay to the stimulus of about 15 ms, and a repolarization within 30 to 300 ms. The power-stroke direction of the cirri-beat reversed from backward to forward during this response. The receptor current rose to an average amplitude of 1.4 nA with a speed of 0.1–0.3 nA/ms and decayed with a single exponential time course with a time constant between 7 and 9 ms. Similar current-reversal potentials, after substitution of extracellular Ca²⁺ by Mg²⁺ and vice versa, indicate that the mechanically activated conductance is identical for Ca²⁺ or Mg²⁺. The current can be carried by Ba²⁺ as well, but not by K⁺ or Na⁺. Decirriation experiments have shown that the mechanosensitivity is located within the soma membrane.Abbreviations EASW artificial sea-water adapted for electrophysiology - EGTA ethylene glycol-O O-bis(2-aminoethyl)-N,N,N,N-tetraacetic acid - NMDG N-methyl-D-glucamine - TEA tetraethyl ammonium - V _m membrane potential - g _X conductance for the ion X - D600 Methoxyverapamil     

Relations stimulus-réponse dans le cas d'un mécanorécepteur d'insecte à adaptation totale

A study of the relations between the applied stimulus, the receptor potential, and the propagated action potentials has been undertaken in the phasic mechanoreceptor of the leg of the locust to establish, subsequently, the transfer function of the system. In the present paper, only rectangular steps were applied to the external structure.The relations have been studied during the dynamic phase of the stimulus application and during the stationary phase in the course of which the receptor shows complete adaptation.The dynamic component of the receptor potential increases with the logarithm of the intensity of the stimulation, to a maximum which is not imposed by a mechanical obstacle. Then, the receptor potential decreases under the maintained stimulus as a power function of the time. During the dynamic phase it is impossible to establish a linear relationship between the amplitude of the receptor potential and the instantaneous frequency of the spikes which is limited, even for small stimulations, by the refractory period.During the adaptation phase, which begins more rapidly for the instantaneous frequency than for the receptor potential, the relation between the two phenomena becomes linear, and thus the decrease in frequency is also a power function.Electrical stimulations of the initiation site of action potentials show that the former adapts to a constant current as an exponential function of time. The adaptation to a mechanical stimulus and the adaptation to an electrical stimulus are graphically compared.     

Soil solute concentration and water uptake by single lupin and radish plant roots

Application of computer assisted tomography to gamma and X-ray attenuation measurements and Na⁺-LIX microelectrodes were used to determine the spatial distributions of soil water content and Na⁺ concentrations respectively near single roots of eighteen day old lupin and radish plants. These quantities were monitored at root depths of 3, 6 and 9 cm and at zero, 2, 4, 6, and 8 hour intervals from the diurnal commencement of transpiration. The plants were subjected to two levels of transpirational demand and five Na⁺ soil solution concentration levels. Water extraction rates for the lupin and radish roots increased continuously with time but were substantially reduced with increasing Na⁺ concentration in the treatment. Water uptake was uniform along the length of the essentially constant diameter lupin roots but decreased along the tapering radish roots as the diameter and hence the surface area per unit length of the roots decreased. The accumulation of Na⁺ at the root surfaces of both plants increased gradually with time in a near linear fashion and was slightly higher under the higher transpiration demand. These increases were not exponential as would be expected with non-absorption by the roots and this is considered to be due to back diffusion at the relatively high water contents used. At these water contents matric potentials had a much smaller influence on transpiration than osmotic potentials. The relationships between leaf water potentials (Ψ₁) and osmotic potentials at the root surfaces were linear with the decreases in Ψ₁ almost exactly reflecting the decreases in Ψ_π indicating rapid plant adjustment. Leaf water potentials decreased progressively with time and the relationships between leaf water potential and the transpiration rate were also linear supporting the suggestion of constant plant resistances at any given concentration.     

Membrane Characteristics of the Canine Papillary Muscle Fiber

Passive and active responses to intracellular and extracellular stimulation were studied in the canine papillary muscle. The electrotonic potential produced by extracellular polarization with the partition chamber method fitted the time course and the spatial decay expected from the cable theory (the time constant, 3.3 msec; the space constant, 1.2 mm). Contrariwise, spatial decay of the electrotonic potentials produced by intracellular polarization was very short and did not fit the decay curve expected for a simple cable, although only a small difference of time course in the electrotonic potentials produced by intracellular and extracellular polarizations was observed. A similar time course might result from the fact that when current flow results from intracellular polarization, the input resistance is less dependent on the membrane resistance. The foot of the propagated action potential rose exponentially with a time constant of 1.1 msec and a conduction velocity of 0.68 m/sec. The membrane capacity was calculated from the time constant of the foot potential and the conduction velocity to be 0.76 µF/cm². The responses of the papillary muscle membrane to intracellular stimulation differed from those to extracellular stimulation applied with the partition method in the following ways: higher threshold potential, shorter latency for the active response, linearity of the current-voltage relationship, and no reduction in the membrane resistance at the crest of the action potential during current flow.     

Transient and persistent tetrodotoxin-sensitive sodium currents in squid olfactory receptor neurons

Squid olfactory receptor neurons are primary bipolar sensory neurons capable of transducing water-born odorant signals into electrical impulses that are transmitted to the brain. In this study, we have identified and characterized the macroscopic properties of voltage-gated Na⁺ channels in olfactory receptor neurons from the squid Lolliguncula brevis. Using whole-cell voltage-clamp techniques, we found that the voltage-gated Na⁺ channels were tetrodotoxin sensitive and had current densities ranging from 5 to 169 pA pF⁻¹. Analyses of the voltage dependence and kinetics revealed interesting differences from voltage-gated Na⁺ channels in olfactory receptor neurons from other species; the voltage of half-inactivation was shifted to the right and the voltage of half-activation was shifted to the left such that a “window-current” occurred, where 10–18% of the Na⁺ channels activated and did not inactivate at potentials near action potential threshold. Our findings suggest that in squid olfactory neurons, a subset of voltage-gated Na⁺ channels may play a role in generating a pacemaker-type current for setting the tonic levels of electrical activity required for transmission of hyperpolarizing odor responses to the brain. Accepted: 1 October 1998     

Ion gradient and photoreceptor sensitivity

Summary In superposition eyes of moths (Deilephila elpenor, Manduca sexta) and a neuropteran (Ascalaphus macaronius) receptor excitation and adaptation depend on the limited store of available ion charges in the extra-cellular space.Na⁺ is the main charge carrier necessary for excitation. Receptor mass responses (measured by extracellular electrodes) can, however, be elicited also after complete removal of Na⁺ from the extra-ommatidial space. Responses are also obtained when Na⁺ is partially replaced by Ca⁺⁺ or Mg⁺⁺. These results suggest that the extra-ommatidial space is separated from the intra-ommatidial (rhabdomeric) extra-cellular space. Ion flow between these spaces is slow. The ion exchange during illumination probably takes place over the rhabdomeric part of the photoreceptor membrane.The Na⁺ concentration in the rhabdomeric extracellular space is about 150 mM, as shown by flame photometry. This result supports the assumption that this space is the common Na⁺ store for the three receptor types (UV, green, violet).Positive (optic lobe positive) DC polarization of the retina causes a reduction in the amplitude of the mass responses to light. After cessation of the polarization the amplitude increases with a time constant similar to that after strong illumination. Negative polarization accelerates the rate of increase in amplitude during dark adaptation, while positive polarization retards the increase. These results suggest that one important factor determining the time course of sensitivity increase during dark adaptation is the reestablishment of the Na⁺ gradient over the rhabdomeric membrane.There is a strong electric coupling between the three receptor types in the ommatidium, since all receptors share the same limited intra-ommatidial extra-cellular ion store. Strong illumination of one receptor reduces the ion gradient (extra- to intracellular) also for the other receptors (not absorbing the light). Thereby the sensitivity of the non-illuminated receptors is also reduced. The electric coupling probably improves the wavelength discrimination by the receptors, by keeping almost constant the relative sensitivity of all three receptor types during shortlasting strong selective adaptation of one or two receptor types.The time course of the resistance change over the receptor membrane (measured extra-cellularly) during and after illumination, suggests that during illumination the Na⁺ influx is reduced, and that the ion gradient over the receptor membrane is at least partially re-established already during the illumination.We are grateful to Dr. A.H. Baumhover (United States Department of Agriculture, Agricultural Research Service, Oxford, North Carolina 27565, USA) for supplying theManduca pupae. The work was supported by the Deutsche Forschungsgemeinschaft, SFB 114 and Rezeptorphysiologie; and by Karolinska Institutets Fonder.     

Inward Rectifying K Channels in the Plasma Membrane of Arabidopsis thaliana

Ion channels in the plasma membrane of protoplasts isolated from cultured cells of Arabidopsis thaliana were studied by means of the patch-clamp technique applied in the whole-cell configuration. In some protoplasts, depolarizing pulses and, in other protoplasts, hyperpolarizing pulses elicited time-dependent currents; both kinds of current were only rarely observed in the same protoplast. The hyperpolarization-activated inward rectifying currents, the focus of this paper, appeared to be due to the relatively slow opening of channels (activation time constant = 150 to 300 milliseconds), which closed at positive potentials. The reversal potential of this current, measured in the presence of different ion concentrations (symmetrical or asymmetrical K⁺ and Cl⁻ or gluconate), was always close to the electrochemical equilibrium potential of K⁺. The currents were inhibited by 10 millimolar tetraethylammonium, a K⁺ channel blocker. These data show that the hyperpolarization-activated currents flow through K⁺ channels, which can provide a pathway for the passive diffusion of K⁺ down its electrochemical gradient.     

Induction of TRPV1 desensitization by a biased receptor agonist

Selective suppression of hyperactive sensory neurons is an attractive strategy for managing pathological pain. Blocking Na⁺ channels to eliminate action potentials and desensitizing transduction channels can both reduce sensory neuron excitability. The novel synthetic vanilloid ligand cap-ET preserves agonist activation of intracellular Ca²⁺ signals and large organic cation transport but loses effective electric current induction. Cap-ET can therefore be used to deliver the membrane impermeable Na⁺ channel blocker QX-314 to substantially inhibit voltage-activated Na⁺ currents. We explored, besides facilitating entry of organic cationic therapeutics, whether cap-ET can also produce receptor desensitization similar to the natural agonist capsaicin. Using the YO-PRO-1 based fluorescent dye uptake assay, we found that cap-ET effectively triggered Ca²⁺ dependent desensitization of TRPV1 when the receptor was pre-sensitized with the surrogate oxidative chemical phenylarsine oxide (PAO), suggesting an alternative use of permanently charged cationic capsaicinoids in differential neuronal silencing.