Oscillations of the Membrane Potential of Pulvinar Motor Cells In Situ in Relation to Leaflet Movements of Desmodium motorium

The ultradian rhythmic movement of the lateral leaflets of Desmodiummotorium is accompanied by rhythmic changes of the extra- andintracellular electrical potentials in the pulvinus, which aremeasured in situ in the pulvinus against the bathing solutionof the petiole. Extra- and intracellular potentials oscillatewith 180'b0 phase difference to each other, as shown by simultaneousmeasurements of both types of potentials in the abaxial partof the pulvinus. Light-induced changes of these potentials movein opposite directions. The in situ membrane potential of themotor cells of the pulvinus was calculated from the differencebetween the extra- and intracellular potentials. It was foundto oscillate between –136 and –36 mV, in phase withthe intracellular and inverse to the extracellular potential.The phase relationship between the leaflet movement rhythm andthe in situ membrane potential rhythm was as follows: downwardmovement is preceded and accompanied by a strong depolarization,upward movement by hyperpolarization. Our results suggest that membrane depolarization in pulvinarmotor cells of Desmodium motorium drives and controls potassiumefflux and hyperpolarization potassium influx via potassiumchannels. Key words: Desmodium pulvinus, leaf movement, pulvinar motor cells, electrical potential     

Studies on Mechano-Perception in Characeae: Effects of External Ca2+ and Cl-

The effects of modification of extracellular concentrationsof Ca²⁺ and C^– on mechano-perception were studied in internodalcells of Chara corallina. Cells were stimulated by droppinga piece of glass tubing on them, and the resulting receptorpotentials and action potentials were analyzed. When the Ca²⁺concentration was extremely lowered by adding EGTA, the amplitudesof both receptor potentials and action potentials were attenuated,suggesting the involvement of Ca²⁺ channels. However, the possibilityremained that attenuation of the amplitude of the receptor potentialwas caused by modification of membrane characteristics by extremelowering of [Ca²⁺]_o. When the plasma membrane was depolarizedto about 0 mV by adding 100 mM KC1, responses in the negativedirection were induced upon mechanical stimulation. When theplasma membrane was depolarized by adding 50 mM K²SO⁴, responsesin the positive direction were induced. Thus, Cl^– channelsmay be involved in responses induced by mechanical stimulationunder K+-induced depolarization. (Received January 16, 1996; Accepted March 25, 1997)     

Studies on Mechanoperception in Characean Cells: Pharmacological Analysis

The mechanisms for generating receptor potentials and actionpotentials upon mechanical stimulation were studied in internodalcells of Chara. Receptor potentials and the subsequent actionpotentials could be generated even when the electrogenic protonpump was inhibited, indicating that the proton pump does notplay a central role in generating receptor potentials and actionpotentials. The involvement of Ca²⁺ and/or Cl^– channelsin both receptor and action potentials was suggested, basedon the equilibrium potentials of these ions across the plasmamembrane. Inhibitors of the Ca²⁺ channel, Cl^– channeland stretch-activated channel could not inhibit generation ofthe receptor potential. These findings suggested that the channelsinvolved in generating the receptor potential are insensitiveto these channel inhibitors, although all inhibitors significantlyinhibited the action potential. (Received July 26, 1996; Accepted November 19, 1996)     

Homeostatic Regulation of Membrane Potential by an Electrogenic Ion Pump against Change in the K+ Concentration of the Extra- and Intra-Organ Perfusion Solutions

The dependence of membrane potentials on changes in the extra-cellularK⁺ concentration [K⁺]_e was investigated in potato tuber sliceswith dripping perfusion, and in growing Vigna hypocotyl segmentswith pressurized intra-organ perfusion methods. Only under anoxiawere the membrane potential of potato tuber slices and the electricpotential difference between the parenchyma symplast and xylem(V_px) of Vigna hypocotyl segments depolarized markedly (46 mVand 42 mV/log[K⁺]_e unit, respectively) with increasing [K⁺]_eabove the critical values. The electric potential differencebetween the parenchyma symplast and organ surface (V_ps of thehypocotyl segments remained nearly unchanged up to 30 mEq [K⁺]_e.Under highly aerobic conditions the membrane potentials wererelatively independent of [K⁺]_e except at very high K⁺ concentrations.V_ps showed even hyperpolarization with the increasing KCl concentrationin the perfusion solution that is not in direct contact withthe surface membrane of the parenchyma symplast. The respiration-dependentelectrogenic components of the membrane potentials regularlyincreased with the increasing [K⁺]_e. A voltage-dependent homeostaticcontrol of membrane potential is discussed. (Received August 13, 1984; Accepted December 21, 1984)     

Pacemaker potentials generated by interstitial cells of Cajal in the murine intestine

Pacemaker potentials were recorded in situ from myenteric interstitial cells of Cajal (ICC-MY) in the murine small intestine. The nature of the two components of pacemaker potentials (upstroke and plateau) were investigated and compared with slow waves recorded from circular muscle cells. Pacemaker potentials and slow waves were not blocked by nifedipine (3 µM). In the presence of nifedipine, mibefradil, a voltage-dependent Ca²⁺ channel blocker, reduced the amplitude, frequency, and rate of rise of upstroke depolarization (dV/dt_max) of pacemaker potentials and slow waves in a dose-dependent manner (1–30 µM). Mibefradil (30 µM) changed the pattern of pacemaker potentials from rapidly rising, high-frequency events to slowly depolarizing, low-frequency events with considerable membrane noise (unitary potentials) between pacemaker potentials. Caffeine (3 mM) abolished pacemaker potentials in the presence of mibefradil. Pinacidil (10 µM), an ATP-sensitive K⁺ channel opener, hyperpolarized ICC-MY and increased the amplitude and dV/dt_max without affecting frequency. Pinacidil hyperpolarized smooth muscle cells and attenuated the amplitude and dV/dt_max of slow waves without affecting frequency. The effects of pinacidil were blocked by glibenclamide (10 µM). These data suggest that slow waves are electrotonic potentials driven by pacemaker potentials. The upstroke component of pacemaker potentials is due to activation of dihydropyridine-resistant Ca²⁺ channels, and this depolarization entrains pacemaker activity to create the plateau potential. The plateau potential may be due to summation of unitary potentials generated by individual or small groups of pacemaker units in ICC-MY. Entrainment of unitary potentials appears to depend on Ca²⁺ entry during upstroke depolarization. pacemaker activity; slow waves; gastrointestinal motility; calcium channel     

Action Spectrum of Light Pulse-Induced Membrane Depolarization in Pulvinar Motor Cells of Phaseolus

In addition to circadian changes in the membrane potential andleaf movement, light applied to the pulvinus causes changesin both the membrane potential and the pulvinar movement inPhaseolus vulgaris L. Even after a short pulse of light, a transientdepolarization of the membrane occurs and leaf movement is observed.Decreases of turgor pressure of the motor cells are always precededby the depolarization. The direction of the leaf movement canbe explained by the decrease of turgor pressure in the motorcells on the irradiated side of the pulvinus. Using the OkazakiLarge Spectrograph at the National Institute for Basic Biology,we determined the action spectrum of the membrane depolarizationinduced by light pulses (30 s) in motor cells of Phaseolus.The pulvinus was left exposed to air during measurement of themembrane potential with microelectrodes. The action spectrumobtained was in the range of 300 to 730 nm. It had the highestpeak at 460 nm with lower peaks at 380 nm and 420 nm. Almostno sensitivity was observed at wavelengths shorter than 360nm and longer than 520 nm. Red and far-red light had no effecton the depolarization of the motor cell. The features of theaction spectrum are almost the same as those of the Blue-Typeresponse in plants. (Received January 9, 1997; Accepted February 14, 1997)     

Studies on Mechano-Perception in Characean Cells: Development of a Monitoring Apparatus

An apparatus to monitor the electrical signal generated in Characells upon mechanical stimulation was developed. An internodalcell was stimulated by dropping a glass tubing with the intensityof the stimulus being controlled by changing either the weightof the glass tubing or the height from which it was dropped.Upon stimulation, a receptor potential was generated with theamplitude being dependent on the stimulus intensity. When thereceptor potential reached a threshold value, an action potentialwas generated. The receptor potential and action potential werecharacterized. The usefulness of this apparatus for analyzingreceptor potentials is discussed. (Received January 29, 1996; Accepted April 16, 1996)     

Electrophysiological characterization of murine HL-5 atrial cardiomyocytes

HL-5 cells are cultured murine atrial cardiomyocytes and have been used in studies to address important cellular and molecular questions. However, electrophysiological features of HL-5 cells have not been characterized. In this study, we examined such properties using whole cell patch-clamp techniques. Membrane capacitance of the HL-5 cells was from 8 to 62 pF. The resting membrane potential was –57.8 ± 1.4 mV (n = 51). Intracellular injection of depolarizing currents evoked action potentials (APs) with variable morphologies in 71% of the patched cells. Interestingly, the incidence of successful, current-induced APs positively correlated with the hyperpolarizing degrees of resting membrane potentials (r = 0.99, P < 0.001). Only a few of the patched cells (4 of 51, 7.8%) exhibited spontaneous APs. The muscarinic agonist carbachol activated the acetylcholine-activated K⁺ current and significantly shortened the duration of APs. Immunostaining confirmed the presence of the muscarinic receptor type 2 in HL-5 cells. The hyperpolarization-activated cation current (I_f) was detected in 39% of the patched cells. The voltage to activate 50% of I_f channels was –73.4 ± 1.2 mV (n = 12). Voltage-gated Na⁺, Ca²⁺, and K⁺ currents were observed in the HL-5 cells with variable incidences. Compared with the adult mouse cardiomyocytes, the HL-5 cells had prolonged APs and small outward K⁺ currents. Our data indicate that HL-5 cells display significant electrophysiological heterogeneity of morphological appearance of APs and expression of functional ion channels. Compared with adult murine cardiomyocytes, HL-5 cells show an immature phenotype of cardiac AP morphology. action potential; ion channel; muscarinic receptor     

Measured and Calculated Transpiration in Trifolium repens under Different Water Potentials

The relationship between transpiration measured gravimetrically,a generalized transpiration equation, and the ratio VPD/r_leafwas investigated in Trifolium repens plants subjected to varyingwater potentials. Dawn leaf water potential was measured witha pressure chamber, leaf diffusion resistance with a diffusionporometer, leaf temperatures with a thermistor, and relativehumidity with an aspirated psychrometer. During drought transpirationrates determined by both methods were quite similar particularlyat the lowest water potentials. After rewatering calculatedrates were somewhat higher than measured ones. It is concludedthat transpiration calculated by the indirect method is a usefuland reasonable estimate of transpiration for single plants undervarying water potentials.     

Resting membrane potential and action potential of Nitella expansa protoplasts

Inside negative membrane potentials were observed for protoplastsobtained from Nitella expansa leaf internodal cells in mediacontaining 1 to 100 mM CaCl₂ using the microelectrode technique.The potential values were less negative than the membrane potentialof intact N. expansa leaf internodal cells. In addition, anaction potential consisting of two components—a fast componentand a slow component—was induced by electrical stimulationfor the protoplasts as well as the intact cells. (Received December 18, 1979; )     

Inositol-1,4,5-trisphosphate Induces Responses in Receptor Neurons in Rat Vomeronasal Sensory Slices

Using the whole-cell mode of the patch-clamp technique, we recordedaction potentials, voltage-activated cationic currents and putativesecond messenger-activated currents in receptor neurons in thevomeronasal sensory epithelium of female rats. The resting membranepotential and input resistance were –45.5 ± 2.5mV (mean ± SEM, n = 39) and 1.5 ± 0.2 G     

Electromotive force of Nitella membrane during excitation

Excitation of the Nitella membrane is analysed by assuming themembrane to be an electromotive force in series with a resistance,both being variables of time and of membrane potential. Duringstep depolarization beyond a threshold, conductance and electromotiveforce increase transiently, finally reaching their respectivesteady state levels. The conductance increase peak is attainedearlier than the peak for electromotive force increase. Wheneverelectromotive force increases beyond the level of clamped membranepotential, the ionic current flows inward. This is consideredto be the origin of the apparent negative resistance characteristicof the excitable membrane. Anodal break response and spontaneousfiring of Nitella membrane are also caused by transient increasesin electromotive force and conductance irrespective of whetherthe membrane potential is being held at its resting level. Thetransient increase in electromotive force reflects changes,like a phase transition, occurring during excitation. (Received May 6, 1968; )     

Action of Mono- and Difluorodinitrobenzene on Induced Electrical Modifications by External pH Changes in Nitella

The action of mono- (FNB) or difluorodinitrobenzene (DFNB) onion permeability is mainly attributed to its interactionwithamino groups of the membrane by dinitrophenylation. Nitella cells were dimtrophenylated at pH 7.3 and the membranepotential and electrical resistance were then measured in acidicor basic solutions. No matter what the pH value was, FNB andDFNB induced a depolarization of the membrane potential andcaused a diminution of resistance. However these effects ofFNB and DFNB were more drastic at alkaline pH and in the presenceof a weak concentration of potassium. Neither the addition of0.1 mM calcium nor the substitution of chlorides by nitratesmodified the DFNB effect. These results are compatible withthe assumption that the DFNB binding to the membrane leads toan augmentation of the negative charges of the membrane bringingabout an increased cation conductance and a modification ofthe affinity of a K⁺/H⁺exchange pump. The transient responseof the membrane potential at the time of dinitrophenylationwas used to roughly estimate the total density of amino groupsof the membrane of Nitella.     

Ionic mechanisms of depolarizing and hyperpolarizing quinine receptor potentials in Paramecium caudatum

The ionic mechanisms of the depolarizing and the hyperpolarizing quinine receptor potentials in the ciliate Paramecium caudatum were examined by using a behavioral mutant strain. The depolarizing receptor potential was induced by stimulating the anterior end of the specimen, and the hyperpolarizing receptor potential by stimulating the posterior end. The amplitude of both the depolarizing and the hyperpolarizing receptor potentials increased linearly with logarithmic increase in quinine concentration applied. Threshold concentration for inducing the depolarizing receptor potential was lower than that for the hyperpolarizing one. The peak level of the depolarizing receptor potential shifted towards the depolarizing direction with increasing external Ca²⁺ concentration while that of the hyperpolarizing receptor potential shifted in the depolarizing direction with increasing external K⁺ concentration. Under voltage-clamp conditions, the specimen produced an inward current in response to anterior stimulation, and an outward current in response to posterior stimulation. Both the peak inward and the peak outward currents showed a linear relationship with membrane potential. Current-voltage relationships of the receptor currents indicated conductance increase during the application of quinine. The depolarizing quinine receptor potential appears to be produced by an activation of Ca²⁺ channels, and the hyperpolarizing quinine receptor potential by an activation of K⁺ channels. Accepted: 3 October 1997     

The Influence of Temperature on a K+-Channel and on a Carrier-Type Transporter in Nitella

Fisahn, J. and Hansen, U-P. 1986. The influence of temperatureon a K⁺ -channel and on a carrier type transporter in Nilella—J.exp. Bot. 37. 440–460. In Nitella, the effects of temperature on membrane potentialand on resistance consist of several components. The evaluationof their associated time-constants measured in linear(ized)temperature responses at a resting potential of–120 mVprovides an approach to their identification. For changes slowerthan c. 1 s, the temperature effect on membrane potential andresistance does not originate from temperature action on theinvolved transporter, but is mediated by signals from temperaturesensitive metabolic processes. In the case of potential, theseprocesses seem to be identical to those which also mediate thelight effect: pH-regulation, and two direct signals from photosynthesis,as indicated by the similarities of the related time-constants( respectively). The temperature effect on resistance displays only one time-constant of 40 sinmost experiments. The related process is unknown. The non-coincidenceof the time-constants of the effect on potential and on resistanceimplies the involvement of a carrier-type transporter (H+-pumpor cotransporter) in the effect on potential, and of a K+channelin the effect on resistance. The K+-channel is identified bythe reversal potential of the effect on membrane potential measuredin cells depolarized or hyperpolarized by an injected electricalcurrent Under these conditions the temperature effect on resistancedominates the effect on potential. Key words: H⁺-pump, K⁺-channel, kinetic analysis, Nitella, oscillation, pH-regulation, reversal, potential, temperature, time-constants     

Changes in the Membrane Properties of Nodal Cells of Chara braunii Induced by Exposure to Artificial Pond Water

Nodal cells of Chara braunii were exposed to artificial pondwater (APW) for a month or more, with removal of larger surroundingcells. The membrane resistance, resting potential, amplitudeof the action potential, and threshold voltage measured fromthe resting level were examined. Exposure to APW had markedeffects on membrane properties. (Received April 2, 0990; Accepted August 24, 1990)     

The electrogenic ion pump revealed by the external pH effect on the membrane potential of Nitella. Influences of external ions and electric current on the pH effect

The influences of external ions and electric current on thepH effect of the membrane potential of Nitella are described.Results are explained in terms of a pH-dependent electrogenicion pump. The reduction of membrane resistance with a pH rise is attributedto the pump current being potential-dependent. (Received July 9, 1974; )     

ACTION POTENTIAL OF NITELLA INTERNODES

The ionic current during a non-propagating action potentialis analysed from the voltage clamp experiments. The shape ofthe action potential of the Nitella internode can be reconstructedfrom the data of the voltage clamp experiments. The N-shapedcurrent-voltage characteristics (I-V curve) of the Nitella membraneis not constant with time as it is in the tunnel diode, butdecays with time, converging finally into a delayed rectificationcurve. The temporal locus of the potential at which each I-Vcurve crosses the voltage axis coincides almost exactly withthe action potential. The membrane resistance which is calculatedfrom the slope of the I-V curve at each intersection with thevoltage axis also changes in parallel to the action potential.Such correlations are found in the Nitella not only in the pondwater, but also in high Na, high Ca or high Mg medium, wherethe shape of the action potential is modified in various ways.It is highly probable that the action potential is a locus ofthe change of the membrane potential so that the net membranecurrent may be maintained at zero after the transient modificationof the membrane structure by stimulation. (Received June 30, 1966; )     

Anode break excitation in Chara membrane

The anode break excitation in Chara membrane was analyzed witha model consisting of an electromotive force (emf) in serieswith a resistance r ( = l/g). The emf was found to shift towarddepolarization during supply of an inward current. Wheneverthe shift of the emf went beyond a threshold depolarization,an action potential was brought about after the end of or evenduring the inward current supply. (Received September 24, 1974; )     

Pressure Effects on Membrane Potentials of Mesophyll Cell Protoplasts and Epidermal Cell Protoplasts of Commelina communis L.

Pantoja, O. and Willmer, C. M. 1986. Pressure effects on membranepotentials of mesophyll cell protoplasts and epidermal cellprotoplasts of Commelina communis L.—J. exp. Bot. 37:315–320. Membrane potentials of epidermal cell protoplasts and mesophyllcell protopiasts of Comnelina communis were measured when theprotoplasts were immobilized in a suction micropipette. Whenzero suction was employed, membrane potentials of both protoplasttypes were near to zero. As suction pressure was increased,membrane potentials became increasingly more negative with gradientsof 14·3 mV/kPa and 10·5 mV/kPa for mesophyll cellprotoplasts and epidermal cell protoplasts, respectively. Theplasma membrane is stretched when suction pressure is appliedto protoplasts and it is considered that this simulates cellturgor pressure which is associated with negative membrane potentialsof intact cells. The results help to explain why some investigatorsobtain positive membrane potentials for protoplasts while othersobtain negative values. The results also indicate that considerablecaution is needed in the interpretation of ion flux data whenprotoplasts are used. Key words: Commelina communis, membrane potentials, pressure, protoplasts