Light-induced changes in membrane potential in Spirogyra

Spirogyra cells exhibited changes in membrane potential whenthey were exposed to light. Cells made chloroplast-free didnot show any light-induced potential change (LPC) upon illuminationwith white light and also monochromatic red (680 nm) and farred (720 nm) light. LPC was observed when the cell containedonly a small fragment of chloroplast, whether the cell had anucleus or not. The magnitude of LPC depended on the amountof chloroplast in the cell. DCMU at 10^–5 M, CCCP at 10^–5 M and DNP at 10^–4M at pH 5.5 suppressed LPC, while CCCP at 1–5 ? 10^–6M, NH₄Cl at 5 ? 10^–2 M and DNP at 10^–4 M at pH 7.0stimulated LPC. PMS at 10^–4 M stimulated LPC and couldinduce LPC which was completely inhibited by DCMU. These factssuggest that LPC is related to noncyclic and cyclic electronflows. The influences of light and dark conditions and various metabolicinhibitors (DCMU, DNP, CCCP, NH₄Cl) on ATP level have been investigated.No significant difference in the ATP level was observed betweencells in the light and dark. DNP at 10^–4 M (pH 5.5) andCCCP at 5 ? 10^–6 M decreased the ATP level significantly,while DCMU and NH₄Cl only slightly. Good correlation was notfound between the total ATP level and LPC in Spirogyra. LPC occurred even when the external medium contained only asingle salt such as KCl, NaCl or CaSO₄. LPC was also recorded in chloroplasts in situ and in vitro.The mode of LPC of chloroplasts was quite different from thatof the cell. On illumination, the chloroplast potential changedvery rapidly and transiently in the positive direction thenrecovered spontaneously to almost the original potential level. Possible causes of LPC are discussed in relation to the electrogenicion pump. ¹ Present address: Department of Botany, Faculty of Science,University of Tokyo, Hongo, Bunkyo, Tokyo 113, Japan. (Received November 9, 1977; )     

Pyruvate Uptake by Mesophyll and Bundle Sheath Chloroplasts of a C4 Plant, Panicum miliaceum L.

Intact chloroplasts were isolated from mesophyll and bundlesheath protoplasts of a C₄ plant, Panicum miliaceum L., to measurethe uptake of [1-¹⁴C]pyruvate into their sorbitol-impermeablespaces at 4?C by the silicone oil filtering centrifugation method.When incubated in the dark, both chloroplasts showed similarslow kinetics of pyruvate uptake, and the equilibrium internalconcentrations were almost equal to the external levels. Whenincubated in the light, only mesophyll chloroplasts showed remarkableenhancement of the uptake, the internal concentration reaching10–30 times of the external level after 5 min incubation.The initial uptake rate of the mesophyll chloroplasts was enhancedabout ten fold by light and was saturated with increasing pyruvateconcentration; K_m and V_max were 0.2–0.4 mM and 20–40µmol(mg Chl)^–1 h^–1, respectively. The lightenhancement was abolished by DCMU and uncoupling reagents suchas carbonylcyanide-m-chlorophenylhydrazone and nigericin. Theseresults indicate the existence of a light-dependent pyruvatetransport system in the envelope of mesophyll chloroplasts ofP. miliaceum. The uptake activity of mesophyll chloroplastsboth in the light and the dark was inhibited by sulfhydryl reagentssuch as mersalyl and p-chloromercuriphenylsulfonate, but thebundle sheath activity was insensitive to the reagents. Thesefindings are further evidence for the differentiation of mesophylland bundle sheath chloroplasts of a C₄ plant with respect tometabolite transport. (Received July 3, 1986; Accepted October 8, 1986)     

Studies of pesticides effects on Chlorella metabolism II. Effect of DCMU on galactolipid metabolism

DCMU (N'-(3,4-dichlorophenyl)-N, N-dimethylurea) was testedfor effects on the metabolism of galactolipids in Chlorellaand chloroplasts isolated from higher plants. In Chlorella,DCMU affected galactolipid synthesis in the light more thanthat of other lipids, but it showed no effect on lipid synthesisin the dark. DCMU did not affect the turnover of galactolipidsin the light. In vitro studies using ¹⁴C-acetate or ¹⁴C-UDP-galactoseas a precursor showed that DCMU had no effect on the synthesisof gross lipid or galactolipids in chloroplasts isolated fromhigher plants. The significance of these observations are discussed. (Received September 21, 1974; )     

Light Activation of NADP-Malate Dehydrogenase in Guard Cell Protoplasts from Vicia faba L

Light-induced swelling of guard cell protoplasts (GCP) from Vicia faba was accompanied by increases in content of K⁺ and malate. DCMU inhibited the increase of K⁺ and malate, and consequently swelling.

Effect of light on the activity of selected enzymes that take part in malate formation was studied. When isolated GCP were illuminated, NADP-malate dehydrogenase (NADP-MDH) was activated, and the activity reached a maximum within 5 minutes. The enzyme activity underwent 5- to 6-fold increase in the light. Upon turning off the light, the enzyme was inactivated in 5 minutes NAD-MDH and phosphoenolpyruvate carboxylase (PEPC) were not influenced by light. The rapid light activation of NADP-MDH was inhibited by DCMU, suggesting that the enzyme was activated by reductants from the linear electron transport in chloroplasts. An enzyme localization study by differential centrifugation indicates that NADP-MDH is located in the chloroplasts, NAD-MDH in the cytosol and mitochondria, and PEPC in the cytosol. After light activation, the activity of NADP-MDH in guard cells was 10 times that in mesophyll cells on a chlorophyll basis. The physiological significance of light-dependent activation of NADP-MDH in guard cells is discussed in relation to stomatal movement.

     

Characteristics of light-induced 515-nm absorbance change in spinach chloroplasts at lower temperatures II. Relationship between 515-nm absorbance change and rapid H+ uptake in chloroplasts after a short flash illumination

The absorbance change at 515 nm induced by a short (7.6 µsec)light flash in spinach chloroplasts was studied at sub-roomtemperatures in relation to rapid H⁺ uptake into chloroplasts. Lowering of temperature caused a marked decrease in the rateof recovery of 515-nm absorbance change after a flash illumination.Initial rate of rapid H⁺ uptake, measured with absorbance changeof bromcresol purple (BCP), was also reduced at lower temperatures,in a parallel fashion. Half-recovery time of the absorbancechange at 515 nm and rise-time of the pH-indicating absorbanceincrease of BCP coincided well at each temperature studied.Values of the calculated activation energy for these two processeswere almost the same. The parallelism between the 515-nm absorbance change and therapid H⁺ uptake after a single flash illumination was also observedwhen the electric field decay and/or H⁺ translocation were acceleratedby ionophorous antibiotics, carbonylcyanide m-chlorophenylhydrazoneor phenazine methosulfate. From these results, it is suggestedthat the rapid H⁺ uptake into chloroplast is chemically coupledto electron transfer and at the same time diffusion- (or transport-)controlled. Membrane potential, reflected in the 515-nm absorbancechange is dissipated with the rapid H⁺ influx. A model for theelectron-transfer-coupled H⁺ translocation involving a plastosemiquinoneloop is presented. Dissipation of the illumination-formed inside-positivemembrane potential by the influx of H⁺ is explained by the model. (Received September 17, 1976; )     

Activation of K+-Channel in Membrane Excitation of Nitella axilliformis

Two processes of the K⁺ channel activation in plasma membraneexcitation are suggested for Nitella axilliformis. One is relatedto the repolarizing process in the action potential and theother to the after-hyperpolarization (AH). Extra- and intracellulartetraethylammonium (TEA⁺) and extracellular Co²⁺ prolonged theaction potential, indicating involvement of K⁺ channel activationin the repolarizing process of the action potential. The following findings showed that AH is caused by K⁺ channelactivation. First, AH was inhibited by extracellular K⁺ andRb⁺ but not by Na⁺ and Li⁺. Second, it was not inhibited byintracellular TEA⁺ but by extracellular TEA⁺. Third, the membraneconductance increased during AH. Generation of AH was dependenton the level of the resting membrane potential [(E_m)_rest] whichis affected by the activity of the electrogenic H⁺ pump. AHwas generated, when (E_m)_rest was more positive than a criticalvalue, which was supposed to be the equilibrium potential forK⁺ across the plasma membrane. Since extracellular Ca²⁺ competed with extracellular TEA⁺ andCo²⁺ in prolonging the action potential, and sometimes in inhibitingAH, Ca²⁺ may be involved in the K⁺ channel activation. (Received June 11, 1983; Accepted September 21, 1983)     

Role of pulvinar chloroplasts in light-driven leaf movements of the trifoliate leaf of bean (Phaseolus vulgaris L.)

Laminar pulvini of bean (Phaseolus vulgaris L.) contain numerouschloroplasts in cells of their motor tissue. The quantitativerelationships of the chloroplast pigments, chlorophyll a andb, ß-carotene, lutein, neoxanthin as well as the xanthophyllcycle carotenoids (violaxanthin, antheraxanthin and zeaxanthin)were similar to those of mesophyll chloroplasts from leafletlaminae. Exposure of pulvinules to light caused deepoxidationof violaxanthin to zeaxanthin, showing that the xanthophyllcycle is functioning. Chlorophyll fluorescence analysis of pulvinulesconfirmed that their chloroplasts are capable of both photosyntheticelectron transport and non-photochemical fluorescence quenching,showing that they build up a considerable transthylakoid protongradient in the light. Application of DCMU to excised pulvinulesand laminar discs, as well as to pulvinules of intact, attachedterminal leaflets blocked electron transport and fluorescencequenching. Application of the uncoupler CCCP to intact pulvinulesalso prevented non-photochemical fluorescence quenching. Therate of movement of the low-light-adapted terminal leaflet inresponse to exposure of its pulvinule to overhead red light(500 µmol m^–2 s^–1) was reduced when the pulvinulewas pretreated with DCMU. The pulvinar response to overheadblue light (50 µmol ^–2 s^–1), which is morepronounced than to red light, was not affected by similar pretreatmentwith DCMU. Pretreatment with CCCP caused a short lag in theresponse to red light, but did not affect its subsequent rate.The results suggest that the pulvinar response to red, but notto blue light, requires non-cyclic electron transport and theresulting generation of ATP Key words: Leaf movements, light, non-cyclic electron transport, Phaseolus, pulvinar chloroplasts     

Cytochrome c activates K+ channels before inducing apoptosis

Cellshrinkage is an early prerequisite for apoptosis. Theapoptotic volume decrease is due primarily to loss ofcytoplasmic ions. Increased outward K⁺ currents have indeedbeen implicated in the early stage of apoptosis in many celltypes. We found that cytoplasmic dialysis of cytochrome c(cyt-c), a mitochondria-dependent apoptotic inducer,increases K⁺ currents before inducing nuclear condensationand breakage in pulmonary vascular smooth muscle cells. Thecyt-c-mediated increase in K⁺ currents tookplace rapidly and was not affected by treatment with a specificinhibitor of caspase-9. Cytoplasmic dialysis of recombinant (active)caspase-9 negligibly affected the K⁺ currents. Furthermore,treatment of the cells with staurosporine (ST), an apoptosisinducer that mediates translocation of cyt-c frommitochondria to the cytosol, also increased K⁺ currents,caused cell shrinkage, and induced apoptosis (determined byapoptotic nuclear morphology and TdT-UTP nick end labeling assay).The staurosporine-induced increase in K⁺ currents concurredto the volume decrease but preceded the activation of apoptosis(nuclear condensation and breakage). These results suggest that thecyt-c-induced activation of K⁺ channels and theresultant K⁺ loss play an important role in initiating theapoptotic volume decrease when cells undergo apoptosis.

     

Ion Effluxes during Excitation of Characeae

Ion effluxes during excitation of Chara and Nitellopsis measuredby conductometry method were compared with results obtainedby two other analytical methods, Ag-AgCl method for Cl^–and ion chromatography method for K⁺. In both species, the averageefflux measured by the conductometry method was very close tothose of K⁺ and Cl^–. (Received May 12, 1986; Accepted June 25, 1986)     

Inhibition of Cl- Channel Activation in Chara corallina Membrane by Lanthanum Ion

We examined a role of Ca²⁺ in the activation of the two majorion channels, i.e., Cl^– and K⁺ channels at the excitationof the characean plasmalemma. The current-voltage relation (I-Vcurve) of the Chara membrane was compared under the ramp voltageclamp condition before and after external application of 20µMof La³⁺ (a Ca²⁺ channel blocker). The transient inward currentcomponent, which is carried mainly by the efflux of Cl^–,disappeared almost completely in about 30 min with La³⁺ treatment.On the other hand, no effect was observed on the late largeoutward current, which is mainly carried by the efflux of K⁺in a large depolarization region (less negative than –50mV). These results suggest that the Cl^– channel in theChara plasmalemma is activated by Ca²⁺ influx, while the K⁺channel is simply activated by depolarization. (Received April 7, 1986; Accepted June 6, 1986)     

Overexpression of human KCNA5 increases IK V and enhances apoptosis

Apoptotic cell shrinkage, an early hallmark of apoptosis, is regulated by K⁺ efflux and K⁺ channel activity. Inhibited apoptosis and downregulated K⁺ channels in pulmonary artery smooth muscle cells (PASMC) have been implicated in development of pulmonary vascular medial hypertrophy and pulmonary hypertension. The objective of this study was to test the hypothesis that overexpression of KCNA5, which encodes a delayed-rectifier voltage-gated K⁺ (Kv) channel, increases K⁺ currents and enhances apoptosis. Transient transfection of KCNA5 caused 25- to 34-fold increase in KCNA5 channel protein level and 24- to 29-fold increase in Kv channel current (I_K(V)) at +60 mV in COS-7 and rat PASMC, respectively. In KCNA5-transfected COS-7 cells, staurosporine (ST)-mediated increases in caspase-3 activity and the percentage of cells undergoing apoptosis were both enhanced, whereas basal apoptosis (without ST stimulation) was unchanged compared with cells transfected with an empty vector. In rat PASMC, however, transfection of KCNA5 alone caused marked increase in basal apoptosis, in addition to enhancing ST-mediated apoptosis. Furthermore, ST-induced apoptotic cell shrinkage was significantly accelerated in COS-7 cells and rat PASMC transfected with KCNA5, and blockade of KCNA5 channels with 4-aminopyridine (4-AP) reduced K⁺ currents through KCNA5 channels and inhibited ST-induced apoptosis in KCNA5-transfected COS-7 cells. Overexpression of the human KCNA5 gene increases K⁺ currents (i.e., K⁺ efflux or loss), accelerates apoptotic volume decrease (AVD), increases caspase-3 activity, and induces apoptosis. Induction of apoptosis in PASMC by KCNA5 gene transfer may serve as an important strategy for preventing the progression of pulmonary vascular wall thickening and for treating patients with idiopathic pulmonary arterial hypertension (IPAH). potassium ion channel; pulmonary hypertension     

Plasma Membrane H+-ATPase in Guard-Cell Protoplasts from Vicia faba L.

The activity of plasma membrane H⁺-ATPase was measured withmembrane fragments of guard-cell protoplasts isolated from Viciafaba L. ATP hydrolytic activity was slightly inhibited by oligomycinand ammonium molybdate, and markedly inhibited by NO₃^–and vanadate. In the presence of oligomycin, ammonium molybdateand NO₃^–, the ATP-hydrolyzing activity was strongly inhibitedby vanadate. It was also inhibited by diethylstilbestrol (DES),p-chloromercuribenzoic acid (PCMB) and Ca²⁺, but slightly stimulatedby carbonyl cyanide m-chlorophenylhydrazone (CCCP). The acitivityhad higher specificity for ATP as a substrate than other phosphoricesters such as ADP, AMP, GTP and p-nitrophenylphosphate; theK_m was 0.5 mM for ATP. The activity required Mg²⁺ but was notaffected by K⁺, and it was maximal around pH 6.8. When guard-cellprotoplasts were used instead of membrane fragments, the ATPaseactivity reached up to 800µmol Pi.(mg Chl)^–1.h^–1in the presence of lysolecithin. These results indicate thatthe guard cell has a high plasma membrane H⁺-ATPase activity. (Received December 23, 1986; Accepted April 28, 1987)     

Studies on frost hardiness in Chlorella ellipsoidea III. Changes in O2 uptake and evolution during hardening and after freeze-thawing

Chlorella ellipsoidea cells at an intermediate stage in theripening phase of the cell cycle were hardened at 3?C. Oligomycin(OGM) and 3-(3,4-dichiorophenyl)-1,1-dimethylurea (DCMU) addedduring hardening in the light inhibited the development of frosthardiness, suggesting that mitochondria and chloroplasts wereinvolved in the hardening process. The O₂-uptake activity in unhardened cells increased duringhardening in the light while the O²-evolution activity decreased,when these activities were measured at 25?C. The increase inO₂ uptake was suppressed by OGM and DCMU and the decrease inO₂ evolution was stimulated by OGM. While the algal hardinessin the dark was very limited, the addition of glucose duringhardening in the dark caused a remarkable development of frosthardiness. These results suggest that mitochondria and chloroplastsclosely interact at low temperature, and the former plays aprincipal role in the hardening process and the latter servesas substrate-donor in the light. The O₂ evolution in cells which survived freezing was remarkablydecreased by freeze-thawing while the O₂ uptake was hardly affected.The freeze-injured chloroplasts were repaired during the followingincubation. OGM inhibited the repair of freeze-injured chloroplasts.From the results, mitochondria seem to change their membranesinto a structure hardier than chloroplasts, and ATP synthesizedby mitochondria seems to be essential for the repair of freeze-injuredchloroplasts. ¹ Present address: Department of Public Health, Faculty of Medicine,Kyushu University, Maidashi 3-1-1, Higashiku, Fukuoka 812, Japan. (Received November 9, 1977; )     

Measurement of Changes in Cytosolic Ca2+ in Arabidopsis Guard Cells and Mesophyll Cells in Response to Blue Light

Phototropins (phot1 and phot2) are blue light (BL) receptorsthat mediate responses including phototropism, chloroplast movementand stomatal opening, and increased cytosolic Ca²⁺. BL absorbedby phototropins activates plasma membrane H⁺-ATPase in guardcells, resulting in membrane hyperpolarization, and drives K⁺uptake and stomatal opening. However, it is unclear whetherthe phototropin-mediated Ca²⁺ increase activates the H⁺-ATPase.Here, we determined cytosolic Ca²⁺ concentrations in guard cellprotoplasts (GCPs) from Arabidopsis transformed with aequorin.Cytosolic Ca²⁺ increased rapidly in response to BL in GCPs fromboth the wild type and phot1 phot2 double mutants, but was mostlysuppressed by an inhibitor of photosynthetic electron flow (DCMU).With depleted external K⁺, we observed another slower Ca²⁺ increase,which was phototropin- dependent. Fusicoccin, a H⁺-ATPase activator,mimicked the effect of BL. The slow Ca²⁺ increase thus appearsto result from membrane hyperpolarization. The slow Ca²⁺ increasewas suppressed by external K⁺ and was restored by blockers ofinward-rectifying K⁺ channels, CsCl and tetraethylammonium,suggesting the preferential uptake of K⁺ over Ca²⁺. Such efficientK⁺ uptake in response to BL was not found in mesophyll cells.Both the fast and the slow Ca²⁺ increases were inhibited byCa²⁺ channel blockers (CoCl₂ and LaCl₃) and a chelating agent(EGTA). These results indicate that the phototropin-mediatedCa²⁺ increase was not observed prior to H⁺-ATPase activationin guard cells and that Ca²⁺ entered guard cells via Ca²⁺ channelsthrough photosynthesis and phototropin-mediated membrane hyperpolarization.     

Significance of Ca2+ and K+ in Micrasterias Growth and Morphogenesis

Significance of Ca²⁺ and K⁺ for the complex morphogenesis ofMicrasterias, which takes place through multipolar tip growth,was investigated. Studies with different external Ca²⁺ concentrationsand Ca²⁺ channel inhibitors LaCl₃ and verapamil indicate thatCa²⁺ and Ca²⁺ channels are essential in the development, whiletreatments with different K⁺ concentrations and K⁺ channel inhibitorTEA demonstrate that potassium or K⁺ channels are not neededin the process, albeit the existence of K⁺ channels. K⁺ is notneeded even for the regulation of turgor pressure, which wasfound to decrease clearly during cell development. The plasmamembrane ATPase inhibitors diethylstilbesterol (DES) and Na-orthovanadatestop morphogenesis and indicate the importance of ion pumpsin the developmental process. Both supraoptimal, external K⁺and Ca²⁺ cause abundant Ca²⁺ precipitate formation in chloroplasts,which shows that chloroplasts are important in regulation ofcytoplasmic Ca²⁺ metabolism and that K⁺ activates the uptakeof Ca²⁺ through Ca²⁺ channels. (Received June 13, 1995; Accepted September 13, 1996)     

Activation of K+ channels induces apoptosis in vascular smooth muscle cells

Intracellular K⁺ playsan important role in controlling the cytoplasmic ion homeostasis formaintaining cell volume and inhibiting apoptotic enzymes in thecytosol and nucleus. Cytoplasmic K⁺ concentration is mainlyregulated by K⁺ uptake viaNa⁺-K⁺-ATPase and K⁺ efflux throughK⁺ channels in the plasma membrane. Carbonyl cyanidep-trifluoromethoxyphenylhydrazone (FCCP), a protonophorethat dissipates the H⁺ gradient across the inner membraneof mitochondria, induces apoptosis in many cell types. In ratand human pulmonary artery smooth muscle cells (PASMC), FCCP opened thelarge-conductance, voltage- and Ca²⁺-sensitiveK⁺ (maxi-K) channels, increased K⁺ currentsthrough maxi-K channels [I_K(Ca)], and inducedapoptosis. Tetraethylammonia (1 mM) and iberiotoxin (100 nM)decreased I_K(Ca) by blocking the sarcolemmalmaxi-K channels and inhibited the FCCP-induced apoptosis inPASMC cultured in media containing serum and growth factors.Furthermore, inhibition of K⁺ efflux by raisingextracellular K⁺ concentration from 5 to 40 mM alsoattenuated PASMC apoptosis induced by FCCP and theK⁺ ionophore valinomycin. These results suggest thatFCCP-mediated apoptosis in PASMC is partially due to anincrease of maxi-K channel activity. The resultant K⁺ lossthrough opened maxi-K channels may serve as a trigger for cellshrinkage and caspase activation, which are major characteristics ofapoptosis in pulmonary vascular smooth muscle cells.

     

Studies on cytochromes in photosynthetic electron transport system I. photoreduction and photo-oxidation of cytochrome b559 by photosystem II in spinach chloroplasts

Light-induced redox-reactions of cytochrome b₅₅₉ in spinachchloroplasts were investigated. Illumination of chloroplastsinduced photoreduction of cytochrorne b₅₅₉ Red light (650 nm)was more effective than far-red light (725 nm), indicating thatthe photoreduction is a photosystem II-mediated reaction. Onaddition of DCMU, the photoreduction was eliminated and a photooxidationof cytochrome b₅₅₉ was observed. The rate of this photooxidationwas faster with photosystem II light than with photo-systemI light. On addition of Mn⁺⁺ the photooxidation was partly suppressed;far-red light became as effective as red light in inducing photooxidationof cytochrome b₅₉₉, in the presence of DCMU and Mn⁺⁺. Ascorbate completely suppressed photooxidation of cytochromeb⁵⁵⁹ In the presence of ascorbate, however, photooxidation wasobserved in the presence of inhibitors or after inhibitory treatmentsof chloroplasts which affected the oxidizing side of systemII. These inhibitors and inhibitory treatments, but not DCMU,decreased the redoxpotential of cytochrome b₅₅₉. Reactivationof Hill reaction in Tris-washed chloroplasts by indophenol-ascorbatetreatment was not accompanied by an abolishment of photooxidationof cytochrome b₅₅₉. A possible mechanism is proposed to account for these reactionsof cytochrome b₅₅₉ in the photosynthetic electron transportin chloroplasts. (Received April 4, 1972; )     

Movements of K+ during Shutting and Opening of the Trap-Lobes in Aldrovanda vesiculosa

K⁺ movements during the shutting and subsequent opening of trap-lobesin Aldrovanda vesiculosa were measured using ⁸⁶Rb as a tracerfor K⁺. Immediately after the shutting, a large amount of ⁸⁶Rbpre-loaded in the trap-lobes was detected in the hollow spaceinside the shut trap. This may indicate that much of the K⁺in the active motor cells leaks out during the shutting, resultingin turgor loss in the cells. ⁸⁶Rb(K⁺) uptake in the trap wasactive. During the opening process, enhanced ⁸⁶Rb uptake wasobserved. The time course of this uptake was similar to thatof the opening of the trap-lobes, and both courses were acceleratedby IAA. Enhanced K⁺ uptake may restore the turgor in activemotor cells. The quantity of K⁺ that moved during the shuttingor opening was estimated as 20% of that in the active motorcells in the open state of the trap-lobes. The K⁺ efflux acrossthe membranes of the active motor cells may be caused by a largeincrease in bulk flow triggered by an action potential, andwas estimated as 6,200 pmol.cm^–2. ¹ This paper is dedicated to the memory of Professor Joji Ashidawho established the physiology of rapid movement in Aldrovandavesiculosa. (Received July 22, 1982; Accepted November 11, 1982)     

Light-induced Activation of Electrogenic H+ Extrusion and K+ Uptake in Elodea densa Depends on Photosynthesis and is Mediated by the Plasma membrane H+ ATPase

In Elodea densa leaves light strongly stimulates electrogenic,K ⁺-dependent, vanadate- and erythrosin B-sensitive H⁺ extrusionand hyperpolarizes the transmembrane electrical potential. Theseeffects of light are suppressed by treatment with DCMU, an inhibitorof photosynthesis, which has no effect on H⁺ extrusion in thedark. Light-induced H⁺ extrusion requires the presence of K⁺in the medium and is associated with increased K⁺ uptake andalkalinization of the cell sap. Light-induced H⁺ extrusion increaseswith increased CO₂ concentration. At constant CO₂ concentration(10⁴ parts 10^–6) the rate of H⁺ extrusion is stronglyenhanced by an increased light intensity up to 30 W m^–2.Different wavelengths, between 400 and 730 nm, induce a significantstimulation of both proton secretion and transmembrane potentialhyperpolarization. The stimulating effects of light on H⁺ extrusion, K⁺ uptakeand cell sap pH are very similar to those induced in the darkby fusicoccin, a toxin known to stimulate strongly ATP-driven,vanadate- and erythrosin B-sensitive H⁺ transport. In the light,the effects of fusicoccin are only partially additive to thoseof light, thus suggesting that the two factors influence thesame system. The identification of this system with the plasmamembrane H⁺-ATPase is indicated by the observed inhibition ofthe effects of either light or fusicoccin by the H⁺-ATPase inhibitorsvanadate and erythrosin B. These data indicate that the activation of electrogenic H⁺ extrusionand of K⁺ uptake by light is mediated by some products of photosynthesis.The mechanism and the possible physiological implications ofthis phenomenon are discussed. Key words: Photosynthesis, H⁺ pump, K⁺ uptake, Elodea densa     

K+ Channel in the Chora Plasmalemma: Estimations of K+ Channel Density and Single K+ Channel Conductance

The electromotive force E and the conductance G of the Characorallina plasmalemma were measured under voltage clamp conditions.In the depolarized voltage range less negative than –60mV, E changed according to the Nerhst equation for K⁺, and Gincreased with the external K⁺ concentration [K⁺]_o and alsowith the depolarization of the membrane potential. This is attributedto the voltage-dependent opening of the K⁺ channels in the largelydepolarized voltage region. The voltage-dependent increase ofG was due to the increase of the number of open K⁺ channelsper unit area. The density of the total K⁺ channels in the C. corallina plasmalemmawas estimated to be about 6.50/(10 µm)2. The single K⁺channel conductance _K changed with the external [K⁺]_o; it was79.3, 86.1, 105.9, 119.0 pS for external [K⁺]_o of 0.2, 0.5,2.0 and 5.0 mu respectively. (Received May 22, 1986; Accepted August 22, 1986)