Conduction Velocity and Blockage of Action Potential in Chara Internodal Cells

Conduction velocity of the action potential in Chara brauniiinternodal cells was 0.21 ?0.05 cms in moist air and 1.5?0.9cms in artificial pond water (APW). The action potential waspropagated at an almost constant velocity along the cell inmoist air except within 0.3 cm from an end of the cell, whereasin APW, the velocity increased to 5.7 ? 2.3 cms within 1.8 cmfrom the end of the cell. When part of the cell was put in moistair and the other part was immersed in APW, conduction of theaction potential in moist air decreased in velocity and sometimesstopped in the vicinity of the boundary between moist air andAPW. Some cells from the plants collected in late autumn towinter generated an action potential which could not propagatein moist air. In these cells, an increase in the threshold andpartial cessation of protoplasmic streaming were observed. ¹Present address: Department of Physiology, Tohoku UniversitySchool of Dentistry, Seiryo-machi, Sendai 980 ²Present address: Biology Laboratory, Kyoritsu Women's University,Hachioji, Tokyo 193, Japan. (Received March 10, 1987; Accepted July 6, 1987)     

Action Potential in Luffa cylindlica and its Effects on Elongation Growth

The action potential of the non-seismonastic plant, Luffa cylindrica,was studied and characterized. Single action potentials of 30to 60 mV were evoked by electrical or cooling stimuli appliedto the stem internode. Action potentials were transmitted withoutdiminution along stimulated internodes shorter than 40 mm inlength. No difference in velocity between acropetal and basipetaltransmission was observed. This behavior is very similar tothat of Mimosa and Biophytum. The velocity of transmission ofaction potentials along internodes of Luffa was dependent upondistance from the point of stimulation; 10 to 20 mm sec^–1upto 40 mm, and 70 mm sec^–1 at around 50 mm from the pointof stimulation. The action potential was not transmitted tothe neighboring internode. Action potentials were also observedin the petiole and leaf. Beyond the threshold stimulus, theamplitude of the transmitted action potential depended on thestrength of the stimulus. The absolute and relative refractoryperiods for the transmission of action potentials were 2 minand 2 to 5 min, respectively. Elongation growth of the stem, recorded using a differentialtransformer, decreased after the generation of a single actionpotential in the upper most internode. This decreased growthrate continued for more than 5 min. (Received March 10, 1986; Accepted June 11, 1986)     

Electrotonic Transmission of an Action Potential between Two Separated Internodal Cells of Chara through a Bridge

Two separated internodal cells of Chara braunii were broughtinto contact with each other longitudinally at their ends andconnected by another pathway composed of a metal bridge beyondthe region of intercellular contact. A conducted action potentialthat arrived at one foot of the bridge electrotonically depolarizedthe other foot of the bridge in the connected cell. The electriccoupling ratio (0.07?0.03), the ratio of the change in the membranepotential of one cell to that of the other cell, was too smallto allow transmission of an action potential. Two cells wereplaced in parallel and connected with two liquid bridges orpools, a' and b'. When the action potential of one cell wasconducted through one connecting pool (pool a)', the other cellwas depolarized electrotonically by the action current via theother connecting pool (pool b'). The coupling ratio was increasedto 0.26?0.07 by the solution bridge, but transmission of theaction potential was rarely observed. Application of 1 mM KC1to pools a' and/or b' slightly improved the frequency of transmissionof the action potential. When pool b' contained 5% urethane,the coupling ratio increased to 0.31?0.08 and transmission ofthe action potential was frequent. (Received August 24, 1989; Accepted March 14, 1990)     

Action Potential in the Trap-lobes of Aldrovanda vesiculosa

The trap of Aldrovanda vesiculosa, an aquatic insectivorousplant, consists of a pair of lobes (trap-lobes) which bordereach other at the midrib. The central portion of the lobe iscomposed of three cell layers, an inner and outer epidermisenclosing a single middle layer of relatively large cells, whereasthe marginal portion consists only of the two epidermal celllayers. Intracellular potentials of these cells were measuredby the microelectrode technique. All the cells of the lobeswere excitable and had identical membrane potentials at rest( –110 mV) and during action (amplitude, 130 mV). Theaction potential of each cell was elicited by bending a sensoryhair, one of many standing on the inner surface of the centralportion, or by injecting an outward current into another cellin the lobe. Action potentials were propagated throughout thetrap-lobes at a rate of about 8 cm/sec. The maximum rising ratewas 2.7 V/sec and the duration of the action potential was 1sec. (Received August 8, 1981; Accepted October 15, 1981)     

Chloride Electrochemical Potentials and Membrane Resistances in Nitella translucens

The chloride electrochemical potential difference between theinside of cells of Nitella translucens and the bathing mediumhas been measured by a direct electrical method employing Ag/AgClelectrodes. The membrane potential has been measured by meansof conventional salt bridge microelectrodes. These data havebeen used to calculate the internal chloride concentration ofthe cells; the mean value obtained was 39 mM. This chlorideelectrochemical potential difference has been short-circuitedthus causing an outward (depolarizing) electric current to flowthrough the cell membrane. The resulting membrane depolarizationhas been measured at two points along the length of the cellenabling the membrane resistance and space constant to be deduced;the respective values obtained were 24.8 Kcm² and 3.0 cm. Itis suggested that these experiments lend additional supportto the hypothesis that during the action potential in the Characeaethere occurs a transient increase in the chloride conductanceof the plasmalemma.     

Changes in Cell Length During Action Potentials in Chara

Changes in cell length during excitation in Chara were recordedsimultaneously with extracellular action potentials. Cells stimulatedin artificial pond water (APW) gave a diphasic change in celllength ; that is, a transient shortening followed by a delayedextension. When a cell was stimulated 1–3 min after aprevious shortening, the extension phase was not evident, andthe amplitude of the shortening was always larger than the previousone. Cells stimulated in moist air gave shortenings with muchlarger amplitudes and much greater durations compared with thosein APW. From such marked differences in both amplitude and durationbetween the two types of shortening it is suggested that theshortening might be caused mainly by an osmotic water movementduring excitation. Net water loss during a single action potentialwas calculated from the cell shortening to be 1.076 nl cm^–2per impulse or 59800 pmol cm^–2 per impulse, which couldbe caused by a local enhancement of ionic concentration justoutside the plasmalemma of about 0.74 mN.     

Electrical rhythmicity and spread of action potentials in longitudinal muscle of guinea pig distal colon

Using simultaneous intracellular recordings, we have characterized 1) electrical activity in the longitudinal muscle (LM) of isolated segments of guinea pig distal colon free to contract spontaneously and 2) extent of propagation of spontaneous action potentials around the circumference of the colon. In all animals, rhythmical spontaneous depolarizations (SDs) were recorded that are usually associated with the generation of action potentials. Recordings from pairs of LM cells, separated by 100 microm in the circumferential axis, revealed that each action potential was phase locked at the two electrodes (mean propagation velocity: 3 mm/s). However, at an increased electrode separation distance of 1 mm circumferentially, action potentials and SDs became increasingly uncoordinated at the two recording sites. No SDs or action potentials ever propagated from one circumferential edge to the other (i.e., 13 mm apart). When LM strips were separated from the myenteric plexus and circular muscle, rhythmically firing SDs and action potentials were still recorded. Atropine (1 microM) or tetrodotoxin (1 microM) either reduced the frequency of SDs or temporarily abolished activity, whereas nifedipine (1 microM) always abolished SDs and action potentials. Kit-positive interstitial cells of Cajal were present at the level of the myenteric plexus and circular and longitudinal muscle. In summary, SDs and action potentials in LM propagate over discrete localized zones, usually <1 mm around the circumference of the colon. Furthermore, in contrast to the classic slow wave, rhythmic depolarizations in LM appear to be generated by an intrinsic property of the smooth muscle itself and are critically dependent on opening of L-type Ca(2+) channels.     

Transmission of action potentials inBiophytum

By cooling or by electrical stimulus an action potential, (60 to 100 mv negative) was propagated throughout the length of a pinna-rachis or a peduncle ofBiophytum sp. A cutting stimulus evoked a series of two to four transmission of actions of action potentials. The transmission did not pass through the base of the leaf or peduncle. The velocity of the transmission in the rachis and penduncle was about 0.2 cm/sec, and no difference in the velocity was found between the acropetal and basipetal directions. In the stimulated site a local response (a negative bulge of potential) was seen with threshold stimulus. The absolute refractory period for the transmission of action potential was estimated at 20 to 50 sec, and the relative one at 30 to 70 sec. The mechanism of the transmission seemed to be similar to that inMimosa pudica.     

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)     

Propagation of Action Potential over the Trap-lobes of Aldrovanda vesiculosa

In the trap-lobes of Aldrovanda vesiculosa, an action potentialwas generated in a cell located at the base of a sensory hairstanding on the margin of central portion of the paired lobes,which spread over this portion within about 40 msec. The electricalcoupling ratio for two adjacent cells in the middle layer ofthe lobes was 0.8. This showed that an action potential generatedin a cell of the trap-lobes must spread electrotonically toadjacent cells. An analysis of an equivalent circuit for thecell injected with current and its neighboring cells in themiddle layer of the lobes showed that the resistances of theplasmalemma, tonoplast and junction between two cells were 11.1,4.7 and 0.56 M, respectively. Numerous plasmodesmata in thejunctional walls of the cells were found by electron microscopy.The low resistance of the junction between cells must be dueto the presence of the plasmodesmata which allows an electrotonictransmission of action potential from cell to cell. (Received January 23, 1982; Accepted April 13, 1982)     

The Genesis and Transmission of Epidermal Potentials of Newt Embryonic Cells in Vivo and in Vitro

The genesis and transmission of action potentials in epidermal cells of a newt ( Cynops pyrrhogaster ) embryo were investigated quantitatively in vivo during development and in vitro in the absence of nerve cells. Typical action potentials, composed of a fast spike followed by a slow action potential, can be recorded from any of the epidermal cells from Stage 24/25 to 35/36. The potential is graded with current intensity, and only the slow component induces transmission to other epidermal cells. The fast spike is found in all epidermal cells from Stage 24/25 to Stage 50; it is abolished by Stage 52. The slow potential disappears at Stage 38 just before or after hatching. The cultured epithelioid explants (epithelioid aggregate) and cultured monolayer cells taken from the presumptive epidermal tissue of the ectoderm of the pregastrula, indicate that sequential changes in the genesis of the dual action potentials are similar to those of the intact embryo. In monolayer cell culture devoid of nerve cells, the epidermal cells, also generate a two-step action potential. Such two-step potentials are characteristic of both ciliated and non-ciliated epidermal cells and occur even during mitotic activity. In contrast, cultured neural plate cells isolated from the neurula generate typical spike-like action potentials.     

Studies on the Development of the Air Pores and Air Chambers of Marchantia paleacea: 1. Light Microscopy

The ontogeny of the air pores and air chambers of Marchantiapaleacea begins with the schizogenous development of protodermalintercellular spaces of the initial apertures, and is completedwith the formation of the air pores and giant sub-epidermalair chambers bearing numerous photosynthetic filaments. Intercellularspace formation commences from the thallus surface and proceedsinwards to the first internal layer of cells. The cells amongwhich spaces develop do not originate from one mother cell.Spaces are formed only in the regions of the intersection ofthe anticlinal walls of three, four, or sometimes more successivederivatives of S₁, S₃ and S₄ segments of the apical cell, oneor two of which have been divided periclinally and the restanticlinally. Protodermal intercellular spaces appear in mostor all the corners of these cells, the anticlinal walls of whichexhibit an opposite disposition. The S₁, S₂, S₃ and S₄ segmentsare produced by definite divisions of a five-sided apical celland by a series of divisions give rise to initial cells of theinternal layers of the thallus and initial cells of the protodermaland sub-protodermal layers. The concept of a quiescent apicalcell cannot be accepted, since dividing apical cells have beenobserved, and the pattern of wall disposition of the thallusapex cannot be explained without the active participation ofthe apical cell. The air chambers are apparently of exogenous origin. They resultfrom the broadening of the bottom of the initial apertures bythe coordination of the rate of anticlinal divisions and growthof the protodermal and sub-protodermal cells surrounding theintercellular spaces of the initial apertures. The ontogenyof the pore rings starts at an advanced stage of air chamberformation not from a mother cell but from the cells which surroundthe closed entrance of the air chamber, by a shift of the planeof division from anticlinal to periclinal. Before the periclinaldivisions a new axis of growth perpendicular to the thallussurface is established in the mother cells of the pore. By a polarized growth into the air chamber followed by periclinaldivisions, the cells of the floor form initial cells of thephotosynthetic filaments. The latter divide again to form singleor branched photosynthetic filaments. Marchantia paleacea, air pore, air chamber     

Spontaneous Action Potential Initiation and Propagation in Regenerating Cell Segments of Acetabularia mediterranea

Thavarungkul, P., Lertsithichai, S. and Sherlock, R. A. 1987.Spontaneous action potential initiation and propagation in regeneratingcell segments of Acetabularia mediterranea.—J. exp. Bot.38: 1541–1556. The spontaneous action potentials which occur during the regenerationof anucleate isolated stalk segments (ISS's) of the unicellularalga Acetabulana mediterranea have been studied using a novelnon-invasive technique. This involved measurement of spatialsamples of the time dependent potential in the external medium(sea water) and an inverse transformation to give the currentdistribution at the cell surface. The initiation region andsubsequent propagation (if any) of each action potential wasthen analysed from the computed transmembrane currents. Theresults showed that the occurrences of the spontaneous actionpotentials followed a rhythm which had a period of approximately24–30 h. These action potentials initiated more frequentlyat the ends of the ISS's than in the middle region. Our resultsshowed no firm correlation between the regions of action potentialinitiation and the site of the regenerating apex. Generallyboth propagating and non-propagating action potentials wereobserved in the same ISS, the ratio of which seems to dependon the length of the cell. The analysed initiation sites andthe propagating behaviour of the spontaneous action potentialgenerated by the ISS's during the regeneration process do notprovide any clear support to the symmetry breaking role of thepropagating action potentials as proposed by some workers. Key words: Acetabularia, regeneration, action potential     

The genesis and transmission of epidermal potentials in an amphibian embryo

The genesis and transmission of action potentials in epidermal cells of the newt (Cynops pyrrhogaster) embryo were investigated with special reference to cellular differentiation during development. Typical action potentials can be recorded from any of the epidermal cells at Stage 31. These potentials consist of a fast spike (18 msec) followed by a slow component (164 msec). The potential is graded with current intensity, and only the slow component initiates action potentials in adjacent cells and induces a transmission to other cells. The fast spike was found in all epidermal cells throughout the embryonic stages examined (Stages 26–47). The slow potential, however, appears at Stage 28, persists until Stage $\text{36}\text{37}$ just before hatching and then disappears at Stage $\text{38}\text{42}$. Electrical recordings from traumatic embryos (embryos without neural crest cells) or from cultured epidermal cell masses isolated from the pregastrula or the ventral region of the neurula, were compared with the intact embryo. No differences were observed in either the form of the action potential or its transmission. Thus these action potentials appear to be derived from epidermal cells, and are not of nervous origin. Evidence suggests that the transient establishment of excitable membranes in epidermal cells during differentiation is closely related to neural cell differentiation.     

The Cellular Pathway of Short-distance Transfer of Photosynthates and Potassium in the Elongating Stem of Phaseolus vulgaris L. A Structural Assessment

The potential cellular pathway of radial transfer of photosynthateand potassium delivered in the phloem to the elongation zone(apical 0.5–2.5 cm) of internode 2 ofPhaseolus vulgarisL. seedlings was elucidated. This was achieved using ultrastructuralobservations of the cell types that constitute the radial pathwayand estimates of potential sucrose and potassium fluxes throughthe cross-sectional area of interconnecting plasmodesmata andacross the plasma membrane surface areas of selected cell types.The investigation relied on predicting the relative roles ofthe mature and developing sieve elements as conduits for theaxial delivery of solutes to the elongation zone. In turn, thesepredictions led to formulation of two transport models whichwere subsequently evaluated. It was found that unloading ofsucrose and potassium from the protophloem sieve elements cannotbe through the symplast due to the absence of plasmodesmata.On the other hand, mature metaphloem sieve element-companioncell complexes have the potential capacity to unload eitherthrough the stem symplast or apoplast. The potential symplasticroute is proposed to be via the companion cells to the adjacentlarge phloem parenchyma cells. Continued radial transfer couldoccur either by exchange to the stem apoplast from the largephloem parenchyma cells or continue in the symplast to the groundtissues. It was further predicted that sucrose utilized forthe development of the procambial/small phloem parenchyma cellscould be delivered axially by them and not by the mature sieveelements. Phaseolus vulgaris ; apoplast; elongating stem; photosynthates; potassium; transport; symplast     

ELECTROPHYSIOLOGICAL STUDIES ON NITELLA CELLS, WITH SPECIAL REFERENCE TO THE ELECTRIC RESISTANCE

Electrical properties of Nitella (Tolypella) cells were studied,with special reference to their electric resistance The resistanceof the protoplasm, as well as the potential of the inside ofthe cell, was measured by inserting capillary electrode intothe cell, which had been sealed with vaseline externally atthe middle so that the current across the protoplasm could bereadily determined. The characteristics of the rectification and impedance weremeasured The electric current was found to flow across the protoplasmiclayer more easily from the sap to the external water than inthe reverse direction. The resistance and the capacitance ofthe cell varied with the a.c frequencies used for the measurement Simultaneous measurements were made of the action potential,the action current, and the resistance change of the protoplasmin order to determine the relationship between them An intimateparallelism between the time courses of these three phenomenawas discovered. An attempt was made to interpret the resultsby taking into account the dissociation of ions in the protoplasm. ¹Present address: Japan Atomic Energy Research Institute, Tokai,Ibaraki Prefecture. (Received May 8, 1962; )     

Action Potentials in Lupinus angustifolius L. Shoots: IV. APPLICATION OF THERMAL STIMULI AND INVESTIGATIONS ON THE CONDUCTION PATHWAYS OF THE EXCITATION

The character of action potentials induced by thermal or electricalstimulation and their blocking by removal of a ring of tissuefrom the shoot as well as the spread of these potentials inthe Lupinus shoot were investigated. The appearance of anothertype of excitation was noted as a graded response occurringlocally. This potential was termed graded potential (GP). Itappears in tissues in which action potentials (AP) are not observed. It is proposed that two interrelated types of reaction to stimulioccur in various types of plant tissue.     

Anatomy and physiology of identified wind-sensitive local interneurons in the cricket cercal sensory system

1. A group of wind sensitive local interneurons (9DL Interneurons) in the terminal abdominal ganglion of the cricket Acheta domesticus were identified and studied using intracellular staining and recording techniques. 2. The 9DL interneurons had apparent resting potentials ranging from -38 mV to -45 mV. At this membrane potential, these cells produced graded responses to wind stimuli; action potentials were never observed at these resting potentials. However, when the 9DL interneurons were hyperpolarized to a membrane potential of approximately -60 mV, a single action potential at the leading edge of the wind stimulus response was sometimes observed. 3. The wind stimulus threshold of the 9DL interneurons to the types of stimuli used in these studies was approximately 0.01 cm/s. Above this threshold, the excitatory responses increased logarithmically with increasing peak wind velocity up to approximately 0.5 cm/s. 4. The 9DL interneurons were directionally sensitive; their response amplitudes varied with wind stimulus orientation. 9DL1 cells responded maximally when stimulated with wind directed at the front of the animal. The apparent peak in directional sensitivity of the 9DL2 interneurons varied between the side and the rear of the animal, depending upon the site of electrode penetration within the cell's dendritic arbor. 5. The locations of dendritic branches of the 9DL interneurons within the afferent map of wind direction were used to predict the excitatory receptive field of these interneurons.     

Action Potentials in Lupinus angustifolius L. shoots

In Lupinus shoots electrical stimulus (DC) produces a voltagewave identical in character with the action potential in stimulatedsimple plant cells or in nerves. These action potentials wereregistered with a Kipp and Zonen Micrograph BD3, calomel electrodes,and an electrometer (input impedance, 10¹⁴ ). The wave form and its amplitude propagation velocity, pointof initiation, threshold value of the stimulus, and refractoryperiod were considered; the interdependence between these parametersand the magnitude and manner of application of the electricalstimulus are discussed. An analogy with Pflüger's laws was found.     

Studies on the Hill reaction of membrane fragments of blue-green algae IV. Garotenoid photobleaching induced by photosystem II action

Carotenoid photobleaching induced by the action of photosystemII was investigated with membrane fragments of Anabaena cylindricaand A. variabilis. Carotenoid photobleaching occurred only when ferricyanide and/orCCCP was present in the reaction mixture. Maximum velocity ofthe reaction was obtained in the presence of ferricyanide andCCCP. Difference spectra (light minus dark) indicated that afast carotenoid photobleaching was accompanied by a slow chlorophyllbleaching. The pattern of the difference spectra was identicalto that reported by Yamashita et al. (1) with spinach chloroplasts.The reaction was DCMU-sensitive, though a portion of the activitywas insensitive to DCMU when ferricyanide was present in thereaction mixture. The effect of CCCP on stimulation of carotenoidphotobleaching showed the same function against CCCP concentrationas that on inhibition of DPIP-photoreduction with DPC. Carotenoidphotobleaching was stimulated by ferricyanide and suppressedby ferrocyanide; thereaction rate was reduced fifty percentwith a ferri- and ferrocyanide mixture giving 510 mV redox potential(pH 7.5). Benzoquinone was stimulatory, but DPIP had no effect. Incubation of membrane fragments in a dilute buffer inactivatedthe Hill reaction but neither the DPC-supported DPIP-photoreductionnor carotenoid photobleaching in A. variabilis. In A. cylindrica,incubation inactivated all three reactions. Inactivation ofthe latter two reactions followed the same kinetics. (Received August 31, 1972; )