Both action potentials and variation potentials induce proteinase inhibitor gene expression in tomato

Tomato plants (Lycopersicon esculentum) accumulate proteinase inhibitor 2 (pin2) mRNA in response to insect attack, crushing and flaming in leaves distant from those treated. Most earlier work suggests that the systemic wound signals are chemical; here we try to determine whether electrical or physical (hydraulic) signals can also evoke pin expression. We used a mild flame to evoke a systemic hydraulic signal and its local electrical aftermath, the variation potential (VP), and we used an electric stimulus to evoke a systemic electrical signal, the action potential (AP). We determined the kinetic parameters of both the VP and AP. Flame-wounded plants essentially always exhibited major electrical responses throughout the plant and a several-fold increase in pin2 mRNA within 1 h. Electrically stimulated plants that generated and transmitted a signal (AP) into the analyzed leaf exhibited similarly large, rapid increases in pin2 mRNA levels. Plants which generated no signal, or signals of just a few microvolts, had unchanged levels of pin2 mRNA. Since the AP and VP both arrived in the receiving leaf before accumulation of pin2 mRNA began, we conclude that, in addition to the previously shown chemical signals, both hydraulically induced VPs and electrically induced APs are capable of evoking pin2 gene expression.     

Glutamate induces series of action potentials and a decrease in circumnutation rate in Helianthus annuus

Reports concerning the function of glutamate (Glu) in the electrical and movement phenomena in plants are scarce. Using the method of extracellular measurement, we recorded electrical potential changes in the stem of 3‐week‐old Helianthus annuus L. plants after injection of Glu solution. Simultaneously, circumnutation movements of the stem were measured with the use of time‐lapse images. Injection of Glu solution at millimolar (200, 50, 5 mM) concentrations in the basal part of the stem evoked a series of action potentials (APs). The APs appeared in the site of injection and in different parts of the stem and were propagated acropetally and/or basipetally along the stem. Glu injection also resulted in a transient, approximately 5‐h‐long decrease in the stem circumnutation rate. The APs initiated and propagating in the sunflower stem after Glu injection testify the existence of a Glu perception system in vascular plants and suggest its involvement in electrical, long‐distance signaling. Our experiments also demonstrated that Glu is a factor affecting circumnutation movements.     

局部灼伤水田芥引起的电波传递以维管系统为主要通道

局部灼伤水田芥（ＮａｓｔｕｒｔｉｕｍｏｆｆｏｃｉｎａｌｅＲ．Ｂｒ．）叶片可引起动作电波、变异电波和持续电波震荡在茎中传递。本实验举出电波传递的主通道为维管系统。由于水田芥的叶序为２／５,灼伤叶片发出的电波首先传递到其维管束直接联接的第五叶片,随后才传递给临近部位的叶片。若把灼伤叶片和茎节连接的维管束切断,则茎中的电波传递即被阻止。用胞内电极可以检测到基层组织的薄壁细胞也有电波出现。     

No relation between drought stress and ethylene production in Norway spruce

The relationship between water availability and ethylene production was studied in 24‐year‐old trees of Norway spruce [ Picea abies (L.) Karst.] growing on experimental plots with different water availability. Ethylene and oxygen were collected from the cambial/xylem region and heartwood in the stem using non‐destructive sampling methods. Xylem sap flow was measured in stems using a heat‐balance technique. Pre‐dawn water potential of shoots was used to assess the water status of the trees. Growth was calculated from increments in stem basal area. The highest ethylene concentrations were found in irrigated trees that also, as compared to the other treatments, showed the most rapid flow rate of sap, the highest pre‐dawn water potential, and the most rapid growth. By contrast, the lowest ethylene concentrations were measured in trees to which artificial drought was induced. Such trees also showed the lowest water transport, lowest water potential and relatively slow growth. Thus, no signs of drought‐induced ethylene production were found in this study, contrary to the general contention of a positive relation between drought stress and ethylene production.     

Cyclopiazonic acid disturbs the regulation of cytosolic calcium when repetitive action potentials are evoked in Dionaea traps

Evoking of action potentials (APs) in the trap of Dionaea muscipula Ellis at intervals shorter than 20 s caused a gradual decrease in the amplitude of the APs. At longer intervals the amplitude was constant. The calcium ionophore A23187 (1 μM) caused a considerable decrease of AP amplitude. Pretreatment of a segment of the Dionaea trap with cyclopiazonic acid (CPA), which is a specific inhibitor of the Ca²⁺-ATPase in the sarcoplasmic seticulum of animal cells and in ER vesicles isolated from plant cells, only slightly affected the amplitude when APs were evoked every 10 min; however, it caused a considerable decrease in the amplitude when the stimulation was repeated every 2 min. Assuming that APs increase the concentration of cytosolic Ca²⁺ and the amplitude of AP depends on the gradient of Ca²⁺ across the plasma membrane, the effect of CPA on the AP amplitude indicates that CPA inhibits the sequestration of Ca²⁺ in Dionaea cells.     

Action potentials in fast- and slow-twitch mammalian muscles during reinnervation and development

Action potentials (APs) were recorded from the extrajunctional membrane of surface fibers of the fast-twitch extensor digitorum longus (extensor) and the slow-twitch soleus muscles of adult rats. APs of the extensor muscle had a significantly faster rate of rise and fall, as well as a shorter duration, than those of the soleus. In addition, the overshoot of APs and the resting membrane potential was greater for the extensor. Whereas the soleus produced only one AP regardless of the stimulus duration, the number of extensor responses was directly proportional to the stimulus duration. This repetitive activity was greatly reduced by a concentration of tetrodotoxin (TTX) as low as 5 X 10(11) g/ml. Within 8 d after crush of the nerves to these two muscles, all differences in AP properties disappeared and both muscles became partially resistant to TTX. Reinnervation brought about a redifferentiation so that differences in AP were again significant at 22 d after nerve crush. However, the rate of rise of extensor APs did not attain normal values even as late as 60 d after nerve crush. APs were found to be the same for extensor and soleus muscles from 12-d-old rats. At 18 d after birth, rate of rise was equivalent to that of adult muscle for the soleus although 50--60 d were required before this parameter was fully mature for the extensor. Nevertheless, APs of the extensor and soleus were clearly differentiated within 25 d after birth. Differences in fast and slow muscle APs are discussed with regard to differences in ion gradients and sarcolemmal conductance.     

Glutamatergic elements in an excitability and circumnutation mechanism

In plants, an electrical potential and circumnutation disturbances are a part of a response to environmental and internal stimuli. Precise relations between electrical potential changes and circumnutation mechanisms are unclear. We have found recently that glutamate (Glu) injection into Helianthus annuus stem induced a series of action potentials (APs) and a transient decrease in circumnutation activity. A theoretical explanation for this finding is discussed here taking into considerations data about the ion mechanism of AP and circumnutation as well as about the metabolic and signaling pathways of glutamate and their possible interactions.Key words: action potential, circumnutation, elongation, glutamate, Helianthus annuus, plant movement     

Action Potentials Evoked by Light in Traps of Dionaea muscipula Ellis

At low light intensities (less than 50 µmol m^–2s^–1) illumination evokes transient depolarization of membranepotential in mesophyll cells of the leaf-trap of Dionaea muscipulaEllis. Darkening causes hyperpolarization approximately symmetricto the response to illumination. The amplitude as well as therate of potential changes depend on light intensity. After exceedinga definite threshold (usually between 50 and 80 µmol m^–2s^–1)the depolarization plays the role of a generator potential andan all-or-none action potential (AP) is released. Switchinglight off in a depolarization phase of an AP does not changeits shape and the amplitude. When the light intensity is increasedto 80–150 µmol m^–2 s^–1 a single lightstimulus triggers two successive APs. The time interval betweenthe two APs decreases with increasing stimulus strength andreaches the minimum between 300 and 400 µmol m^–2s^–1. At higher light intensities the interval increasesagain, and finally only a single AP is triggered. It was shownthat the effect was evoked by light but not by temperature changeaccompanying illumination. An inhibitor of the photosyn-theticelectron transport chain, DCMU, blocked the generator potentialsmediating between light absorption and APs. Residual responsesto light stimuli in plants treated with DCMU had reverse polarityand strongly reduced the amplitudes. (Received September 16, 1997; Accepted January 16, 1998)     

The influence of glutamic and aminoacetic acids on the excitability of the liverwort Conocephalum conicum

Intracellular microelectrode measurements revealed that a resting potential (RP), an action potential (AP) and a calcium component of AP (named voltage transient, VT) can be influenced by glutamic acid (Glu) and aminoacetic acid (glycine, Gly) in the liverwort Conocephalum conicum. In the continuous presence of 5mM Glu or 5mM Gly, the RP hyperpolarized constantly and the plants became desensitized to the excitatory amino acids (Glu or Gly). Under such circumstances, the amplitudes of APs evoked by stimuli other than Glu or Gly grew, as did their calcium components (VTs). The sudden application of 1-15 mM Glu or Gly to a thallus not yet desensitized resulted in an excitation, i.e. a single AP or AP series. Aspartate (Asp) could not substitute for Glu in any way. Simultaneous action of both amino acids acted synergically to trigger APs. The same phenomenon was observed when glycine solution was enriched with N-methyl-D-aspartic acid (NMDA). Gly-induced APs were totally hindered by 1mM D-amino-5-phosphonopentanoic acid (AP5)--an inhibitor of ionotropic glutamate receptors of the NMDA kind. Glu-induced APs could be totally suppressed by 1mM AP5 as well as by 1mM 6,7-dinitroquinoxaline-2,3-dione (DNQX)--an inhibitor of AMPA/KA receptors. DNQX also completely blocked the calcium component of Glu-evoked APs. After DNQX treatment, the only response to Glu was a membrane potential hyperpolarization (like the Glu response in a desensitized plant). It was concluded that the Glu-induced depolarization and hyperpolarization are separate phenomena. The stimulatory effects of both Glu and Gly on liverwort excitability may be the consequences of an activation of a variety of ionotropic Glu receptor subtypes.     

Disturbances of stem circumnutations evoked by wound-induced variation potentials in Helianthus annuus L

The relationship between evoked electrical activity and stem movements in three-week old sunflowers was demonstrated. Electrical potential changes (recorded by Ag/AgCl extracellular electrodes) and time-lapse images (from a top view camera) were recorded and analyzed. A heat stimulus applied to the tip of one of the second pair of leaves evoked a variation potential, transmitted basipetally along one side of the stem. After stimulation, disturbances of circumnutations occurred. They included: changes in the period, disorders in the elliptical shape, and, in some cases, reversion of direction (of movement). We suggest that asymmetrically propagated variation potential induces asymmetric stem shrinking and bending, which strongly disturbs circumnutations. Our results confirm the involvement of electrical potential changes in the mechanism of stem nutations.     

Interweaving and overlapping of evoked potentials

The refractory effect of one stimulus upon the response to a closely following stimulus in a different modality is much less than upon the response to a stimulus in the same modality. It is therefore far more efficient to record responses to stimuli in different modalities concurrently than to record each one separately. We evaluated 2 techniques for concurrent recording. Interweaving involves recording the response to one stimulus in the intervals between recording responses to other stimuli. Overlapping occurs when two or more responses are at times being simulateneously recorded. Interweaving and overlapping reduced the time required to record auditory brain-stem responses, short-latency somatosensory evoked potentials and pattern-reversal visual evoked potentials by a factor of 3 over the time required to record each response separately. Overlapping caused no significant change in the evoked potentials. Depending upon the actual timing schedule, interweaving may distort the evoked potentials if later parts of the response to one stimulus override the evoked potential to a following stimulus. Filtering and randomization of stimulus timing may attenuate the effects of these overriding potentials.     

Influence of segmental and extra-segmental conditioning stimuli on cortical potentials evoked by painful electrical stimulation

This study evaluated the effects of two different types of segmental/extra-segmental conditioning stimuli (tonic muscle pain and non-painful vibration) on the subjective experience (perceived pain intensity) and on the cortical evoked potentials to standardized test stimuli (cutaneous electrical stimuli). Twelve subjects participated in two separate sessions to investigate the effects of tonic muscle pain or cutaneous vibration on experimental test stimuli. The experimental protocol contained a baseline registration (test stimuli only), a registration with the test stimuli in combination with the conditioning stimuli, followed by a registration with the test stimuli only. In addition, the effects of the conditioning stimuli were examined at two anatomically separated locations (segmental and extra-segmental). Compared with the test stimulus alone, the perceived pain intensity and peak-to-peak amplitudes of the evoked potentials were unchanged in the presence of non-painful conditioning stimuli at either location. In contrast, a significant decrease of the perceived pain intensity and peak-to-peak amplitudes was found in the presence of painful conditioning stimuli at the extra-segmental sites. Moreover, the topographic maps of the 32-channel recordings suggested that the distribution of the scalp evoked potentials was almost symmetrical around the vertex Cz in the baseline registration. The evoked potentials were generally decreased during hypertonic saline infusion at the extra-segmental sites, but the distribution of the topographic maps did not appear to change. Vibration has previously been shown to inhibit pain, but in the present study the perceived intensity of phasic painful electrical stimuli was unchanged. The reduced perceived pain intensity and the smaller peak-to-peak amplitude of the evoked potential in the presence of extra-segmental conditioning pain are in accordance with the concept of diffuse noxious inhibitory control.     

The combined effect of Cd2+ and ACh on action potentials of Nitellopsis obtusa cells

Interrelations between the action of acetylcholine (ACh) and cadmium ions (Cd²⁺) on bioelectrogenesis of Nitellopsis obtusa cells were investigated. We analyzed repetitively triggered action potentials (AP), their reproducibility, shape and dynamics of membrane potential after AP induction. ACh significantly increased membrane permeability only at high concentrations (1 mM and 5 mM). Repolarisation level of action potential after the first stimulus was much more positive in all cells treated with ACh as compared to the control. Differences of membrane potentials between points just before the first and the second stimuli were 23.4±.0 mV (control); 40.4±5.9 mV (1 mM ACh solution) and 57.7 ± 8.5 mV (5 mM ACh solution). Cd²⁺ at 20 μM concentration was examined as a possible inhibitor of acetylcholinesterase (AChE) in vivo. We found that cadmium strengthens depolarizing effect of acetylcholine after the first stimulus. The highest velocity of AP repolarization was reduced after ACh application and Cd²⁺strengthened this effect. There were no differences in dynamics of membrane potential after repetitively triggered action potentials in ACh or ACh and Cd²⁺ solutions. This shows that cadmium in small concentration acts as inhibitor of acetylcholinesterase.     

Modeling the interactions between stimulation and physiologically induced APs in a mammalian nerve fiber: dependence on frequency and fiber diameter

Electrical stimulation of nerve fibers is used as a therapeutic tool to treat neurophysiological disorders. Despite efforts to model the effects of stimulation, its underlying mechanisms remain unclear. Current mechanistic models quantify the effects that the electrical field produces near the fiber but do not capture interactions between action potentials (APs) initiated by stimulus and APs initiated by underlying physiological activity. In this study, we aim to quantify the effects of stimulation frequency and fiber diameter on AP interactions involving collisions and loss of excitability. We constructed a mechanistic model of a myelinated nerve fiber receiving two inputs: the underlying physiological activity at the terminal end of the fiber, and an external stimulus applied to the middle of the fiber. We define conduction reliability as the percentage of physiological APs that make it to the somatic end of the nerve fiber. At low input frequencies, conduction reliability is greater than 95% and decreases with increasing frequency due to an increase in AP interactions. Conduction reliability is less sensitive to fiber diameter and only decreases slightly with increasing fiber diameter. Finally, both the number and type of AP interactions significantly vary with both input frequencies and fiber diameter. Modeling the interactions between APs initiated by stimulus and APs initiated by underlying physiological activity in a nerve fiber opens opportunities towards understanding mechanisms of electrical stimulation therapies.     

Collision of two action potentials in a single excitable cell

BackgroundIt is a common incident in nature, that two waves or pulses run into each other head-on. The outcome of such an event is of special interest, because it allows conclusions about the underlying physical nature of the pulses. The present experimental study dealt with the head-on meeting of two action potentials (AP) in a single excitable plant cell (Chara braunii internode).MethodsThe membrane potential was monitored with multiple sensors along a single excitable cell. In control experiments, an AP was excited electrically at either end of the cell cylinder. Subsequently, stimuli were applied simultaneously at both ends of the cell in order to generate two APs that met each other head-on.ResultsWhen two action potentials propagated into each other, the pulses did not penetrate but annihilated (N = 26 experiments in n = 10 cells).ConclusionsAPs in excitable plant cells did not penetrate upon meeting head-on. In the classical electrical model, this behavior is specifically attributed to relaxation of ion channel proteins. From an acoustic point of view, annihilation can be viewed as a result of nonlinear material properties (e.g. a phase change).General significanceThe present results suggest that APs in excitable animal and plant cells belong to a similar class of nonlinear phenomena. Intriguingly, other excitation waves in biology (intracellular waves, cortical spreading depression, etc.) also annihilate upon collision and are thus expected to follow the same underlying principles as the observed action potentials.     

Signal transduction in blue light‐induced oxygen uptake in Chlorella cells

In DCMU‐poisoned wild‐type and in non‐photosynthetic pigment mutant cells of Chlorella kessleri , grown heterotrophically with glucose as a carbon source and with nitrate as sole nitrogen source, the known blue light‐enhanced uptake of oxygen and breakdown of starch were reduced by staurosporine and K252a, both potent inhibitors of protein kinase C. This corresponded to sensitivity to these inhibitors of blue light‐induced uptake of nitrate of such organisms. Cells grown with ammonia as sole nitrogen source responded to short wavelength visible irradiation with an increase in oxygen uptake, and this, too, was inhibited by staurosporine and K252a. However, these cells did not show any blue light‐enhanced uptake of nitrate. From these results, enhanced consumption of oxygen under blue light cannot be a consequence of blue light‐induced protein phosphorylation involved in the light‐dependent uptake of nitrate. However, existence of a specific protein phosphorylation within the process of enhancement of oxygen uptake under blue light is not yet proven by the data. There might be a master reaction that induces both processes independently, or there may be influences of other light‐induced processes which lead to enhanced starch breakdown, thereby supplying the glucose for oxidative degradation.     

The mechanism of propagation of variation potentials in wheat leaves

Here we examined the mechanism of propagation of variation potential (VP) induced by burning in wheat leaves. Participation of hydraulic and chemical mechanisms in VP transmission was analyzed by optical coherent tomography and a radioactive tracer method, respectively. The speed of the hydraulic signal considerably exceeded the VP velocity. Investigation of a chemical substance spreading from the zone of local wounding was based on experimental data for radioactive marker transmission derived with a one-dimensional diffusion equation. The speed of the marker transmission was in accordance with VP velocity. The elimination of the potential transmission of a chemical signal by a timed severing of the leaf between the burn site and the recorded site blocked VP propagation. We suggest that a VP is formed by the transmission of a wound substance, the velocity of which is likely increased by hydraulic wave propagation.     

Light-triggered action potentials in the liverwort Conocephalum conicum

The response to light of a liverwort, Conocephalum conicum L., measured as a change in the resting potential, consists of two stages. The first stage is a slight depolarization dependent on light intensity. This plays the role of a generator potential (GP) which induces the second stage - an action potential of the all-or-none character. Action potentials induced by light and by electrical stimuli have the same properties, i.e. identical time course, propagation velocity, and refractory periods. A summation occurs of sub threshold light stimuli and of light and electrical stimuli. The presence of 5⋅10-⁻⁶ M DCMU cancelled the light response and blocked - by inhibition of the electron transport chain - the mechanism leading to GP generation. However, this effect did not produce any change in the response to electrical stimuli.