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Inhibitors of human two-pore channels (TPC1 and TPC2), i.e., verapamil, tetrandrine, and NED-19, are promising medicines used in treatment of serious diseases. In the present study, the impact of these substances on action potentials (APs) and vacuolar channel activity was examined in the aquatic characean algae Nitellopsis obtusa and in the terrestrial liverwort Marchantia polymorpha. In both plant species, verapamil (20–300 µM) caused reduction of AP amplitudes, indicating impaired Ca2+ transport. In N. obtusa, it depolarized the AP excitation threshold and resting potential and prolonged AP duration. In isolated vacuoles of M. polymorpha, verapamil caused a reduction of the open probability of slow vacuolar SV/TPC channels but had almost no effect on K+ channels in the tonoplast of N. obtusa. In both species, tetrandrine (20–100 µM) evoked a pleiotropic effect: reduction of resting potential and AP amplitudes and prolongation of AP repolarization phases, especially in M. polymorpha, but it did not alter vacuolar SV/TPC activity. NED-19 (75 µM) caused both specific and unspecific effects on N. obtusa APs. In M. polymorpha, NED-19 increased the duration of repolarization. However, no inhibition of SV/TPC channels was observed in Marchantia vacuoles, but an increase in open probability and channel flickering. The results indicate an effect on Ca2+ -permeable channels governing plant excitation.
Mateusz Koselski; Vilmantas Pupkis; Kenji Hashimoto; Indre Lapeikaite; Agnieszka Hanaka; Piotr Wasko; Egle Plukaite; Kazuyuki Kuchitsu; Vilma Kisnieriene; Kazimierz Trebacz. Impact of Mammalian Two-Pore Channel Inhibitors on Long-Distance Electrical Signals in the Characean Macroalga Nitellopsis obtusa and the Early Terrestrial Liverwort Marchantia polymorpha. Plants 2021, 10, 647 .
AMA StyleMateusz Koselski, Vilmantas Pupkis, Kenji Hashimoto, Indre Lapeikaite, Agnieszka Hanaka, Piotr Wasko, Egle Plukaite, Kazuyuki Kuchitsu, Vilma Kisnieriene, Kazimierz Trebacz. Impact of Mammalian Two-Pore Channel Inhibitors on Long-Distance Electrical Signals in the Characean Macroalga Nitellopsis obtusa and the Early Terrestrial Liverwort Marchantia polymorpha. Plants. 2021; 10 (4):647.
Chicago/Turabian StyleMateusz Koselski; Vilmantas Pupkis; Kenji Hashimoto; Indre Lapeikaite; Agnieszka Hanaka; Piotr Wasko; Egle Plukaite; Kazuyuki Kuchitsu; Vilma Kisnieriene; Kazimierz Trebacz. 2021. "Impact of Mammalian Two-Pore Channel Inhibitors on Long-Distance Electrical Signals in the Characean Macroalga Nitellopsis obtusa and the Early Terrestrial Liverwort Marchantia polymorpha." Plants 10, no. 4: 647.
Liverworts are pioneer plants that colonized lands. They had to cope with frequent sea water flooding causing salt stress. The role of vacuoles and in particular slow-activating (SV) channels in the salt stress tolerance was addressed in the present study. A patch-clamp method was used to study sodium fluxes through the tonoplast of the liverwort Conocephalum conicum. The whole-vacuole measurements carried out in a symmetrical Na+ concentration allowed recording of slowly activated outward currents typical for SV channels. In a Na+ gradient promoting an efflux of Na+ from the vacuole, the outward rectifying properties of SV channels were reduced and inward Na+ currents with different inactivation rates were recorded. Single channel analysis proved that a decrease in cytoplasmic Na+ concentration evoked an increase in the open probability of the channels and shifted the activation voltages towards negative values. The number of SV channels recorded at negative voltages was dependent on the vacuolar calcium and decreased at the high concentration of this ion in the vacuole. In some of the tested patches, the channels exhibited a flickering type of activity and two different conductance levels. The role of SV channels in Na+ accumulation during salt stress and its removal after periods of flooding is discussed in the present paper.
Mateusz Koselski; Kazimierz Trebacz; Halina Dziubinska. The role of vacuolar ion channels in salt stress tolerance in the liverwort Conocephalum conicum. Acta Physiologiae Plantarum 2019, 41, 110 .
AMA StyleMateusz Koselski, Kazimierz Trebacz, Halina Dziubinska. The role of vacuolar ion channels in salt stress tolerance in the liverwort Conocephalum conicum. Acta Physiologiae Plantarum. 2019; 41 (7):110.
Chicago/Turabian StyleMateusz Koselski; Kazimierz Trebacz; Halina Dziubinska. 2019. "The role of vacuolar ion channels in salt stress tolerance in the liverwort Conocephalum conicum." Acta Physiologiae Plantarum 41, no. 7: 110.
Mycotoxin stress, one of the currently intensively studied, was stimulated in wheat hypocotyl cells through the application of zearalenone, produced by Fusarium fungi. The differences of reactions in the activation of the SV ion channels between stress-tolerant (Parabola) and sensitive (Raweta) wheat cultivars have been studied by the patch-clamp technique. No impact of zearalenone on SV channel activity was observed in Parabola. However, in the sensitive cultivar, mycotoxin (30 µmol L−1) evoked a significant decrease in the unitary conductance of the channels—from 22 ± 0.4 pS (n = 5) in the control to 18 ± 0.5 pS (n = 7) in zearalenone-treated vacuoles (potential clamped at 100 mV). A combined treatment of zearalenone and selenium (5 µmol L−1 of Na2SeO4) abolished this effect totally, indicating the importance of this element in the defense mechanism of cells under the stress of mycotoxins. The role of differences in the membrane structure between tolerant and sensitive cultivars induced in stress conditions, as one of the main factors in mechanical stability of channels, has been discussed.
Mateusz Koselski; Halina Dziubinska; Kazimierz Trebacz; Apolonia Sieprawska; Maria Filek. The Role of SV Ion Channels Under the Stress of Mycotoxins Induced in Wheat Cells—Protective Action of Selenium Ions. Journal of Plant Growth Regulation 2019, 38, 1255 -1259.
AMA StyleMateusz Koselski, Halina Dziubinska, Kazimierz Trebacz, Apolonia Sieprawska, Maria Filek. The Role of SV Ion Channels Under the Stress of Mycotoxins Induced in Wheat Cells—Protective Action of Selenium Ions. Journal of Plant Growth Regulation. 2019; 38 (4):1255-1259.
Chicago/Turabian StyleMateusz Koselski; Halina Dziubinska; Kazimierz Trebacz; Apolonia Sieprawska; Maria Filek. 2019. "The Role of SV Ion Channels Under the Stress of Mycotoxins Induced in Wheat Cells—Protective Action of Selenium Ions." Journal of Plant Growth Regulation 38, no. 4: 1255-1259.
Microelectrode measurements carried out on leaf cells from Physcomitrella patens revealed that a sudden temperature drop and application of menthol evoked two types of different‐shaped membrane potential changes. Cold stimulation evoked spike‐type responses. Menthol depolarized the cell membrane with different rates. When it reached above 1 mV s‐1, the full response was recorded. Characteristic for the full responses was also a few‐minute plateau of the membrane potential recorded after depolarization. The influence of inhibitors of calcium channels (5 mM Gd3+), potassium channels (5 mM Ba2+), chloride channels (200 μM Zn2+, 50 μM niflumic acid), and proton pumps (10 μM DES), an activator of calcium release from intracellular stores (Sr2+), calcium chelation (by 400 μM EGTA), and phytohormones (50 μM auxin, 50 μM abscisic acid, 500 μM salicylic acid) on cold‐ and menthol‐evoked responses was tested. Both responses are different in respect to the ion mechanism: cold‐evoked depolarizations were influenced by Ba2+ and DES; in turn, menthol‐evoked potential changes were most effectively blocked by Zn2+. Moreover, the effectiveness of menthol in generation of full responses was reduced after administration of auxin or abscisic acid, i.e. phytohormones known for their participation in responses to cold and regulation of proton pumps. The effects of DES indicated that one of the main conditions for generation of menthol‐evoked responses is inhibition of the proton pump activity. Our results indicate that perception of cold and menthol by plants proceeds in different ways due to the differences in ionic mechanism and hormone dependence of cold‐ and menthol‐evoked responses. This article is protected by copyright. All rights reserved.
Mateusz Koselski; Piotr Wasko; Kamila Kupisz; Kazimierz Trebacz. Cold‐ and menthol‐evoked membrane potential changes in the moss Physcomitrella patens : influence of ion channel inhibitors and phytohormones. Physiologia Plantarum 2019, 167, 433 -446.
AMA StyleMateusz Koselski, Piotr Wasko, Kamila Kupisz, Kazimierz Trebacz. Cold‐ and menthol‐evoked membrane potential changes in the moss Physcomitrella patens : influence of ion channel inhibitors and phytohormones. Physiologia Plantarum. 2019; 167 (3):433-446.
Chicago/Turabian StyleMateusz Koselski; Piotr Wasko; Kamila Kupisz; Kazimierz Trebacz. 2019. "Cold‐ and menthol‐evoked membrane potential changes in the moss Physcomitrella patens : influence of ion channel inhibitors and phytohormones." Physiologia Plantarum 167, no. 3: 433-446.
Editorial on the Research Topic Inter-cellular Electrical Signals in Plant Adaptation and Communication Molecular studies suggest that the transition into multi-cellularity took place ~1 billion years ago, after the fungi, animal, and plant lineages had separated (Sanderson, 2003; Peterson and Butterfield, 2005). Although the selective pressures to evolve multi-cellularity are far from understood, comparative studies of two volvocine algae, the colonial Volvox and the unicellular Chlamydomonas suggest that it was a gradual, multi-step process that involved genetic innovations. Among the first multi-cellularity genes were those that encode extracellular matrix proteins that bind cells together, since Volvox has them, but Chlamydomonas does not (Prochnik et al., 2010). The increase in cell number and cell types would not have been possible without additional genetic innovations that made it possible for the thousands, millions, and even billions of cells that compose these multicellular organisms to coordinate their activities and cooperate to produce responses to environmental stimuli. One of the most successful mechanisms to coordinate the activities of distally located cells, tissues, and organs is the electrical impulse, which was first studied in frogs in the 1790s by Galvani and Volta (Galvani, 1791; Pera, 1992). Similar, yet slower electrical impulses were also described in plants a century later by (Burdon Sanderson, 1873), although their relevance was not recognized immediately, being regarded as an anecdotal feature limited to a few exotic species. It is now accepted that most, if not all, plants routinely use electrical and chemical signals that quickly bring into communication distally located cells, yet most aspects of these signals' underlying mechanisms still remain mysterious. With the purpose of promoting this under-researched area of biology and fostering a dialogue between the different perspectives from which it is studied, we invited a diversity of scientists who work on molecular, cellular, and systemic aspects of inter-cellular signals, to contribute their work to this Research Topic. The result is this volume, which contains a diversity of review and original research articles that bring new ideas and elements for reflection, and expand the state-of-the-art of inter-cellular electrical and chemical phenomena, from the ion channel (Ghosh et al.), to the cellular scale (Huang et al.; Niu et al.), and systemic perspectives (Stolarz and Dziubinska; Paulmann et al.). The review by Szechynska-Hebda et al. set forth an interesting hypothesis to explain the propagation of light-induced systemic electrical signals that involves cellular structures in chloroplasts named “stromules.” Stromules are membrane extensions that connect different chloroplasts, and are proposed to operate as electrical platforms that allow the propagation of these light-related signals. Also in this review, the authors discuss a crosstalk between electrical signaling, as well as calcium and ROS waves, phytohormones, gene expression, and pressure changes in the xylem. In addition, they discuss the genetic basis of the electrophysiological responses to plant illumination, focusing on genes that encode ion channels and membrane transporters that are behind these responses. A hypothesis and theory article by Brauchi and collaborators also emphasize the genetic bases plant electrophysiological phenomena, in this case of the inter-cellular signals between different cell types, specifically between vascular and non-vascular cells (Canales et al.). Interestingly, in-silico analyses of tissue-specific expression of various ion channels have led them to suggest that long- and short-distance systemic electrical signals use different cellular routes. Further, they propose a working model for understanding the cellular pathways and mechanisms by which electrical signals travel from the stimulated peripheral areas to the input region of the phloem, and from the phloem to peripheral cells in effector tissues, one that requires the participation of both the apoplast and plasmodesmata. Niu et al. demonstrate the involvement of nitric oxide (NO), Ca2+ and calmodulin (CaM) in adventitious root formation, which is an important aspect of plant strategy to alleviate osmotic stress. They propose a complex molecular mechanism that involves sequenced activation of NO, Ca2+/CaM signaling pathways leading to protection of the photosynthetic apparatus and stimulation of the antioxidant defense system. In another study that also focuses on Ca2+ signaling pathways, Huang et al. ask how subcellular compartmentation of the Ca2+ signal may play a role in imposing specificity on signaling response systems. By targeting a heterologously expressed Ca2+ binding protein to buffer Ca2+ changes in specific subcellular locales, they show that nuclear and cytosolic Ca2+ changes are independently regulated. However, when assaying downstream events such as alterations in gene expression, changes in both compartments are shown to be required for the complex coordinated responses to osmotic and salt stresses. Here, Stolarz and Dziubinska bring new light into the classical problem of the sunflower's circumnutation movement, a phenomenon driven by ion and water fluxes, with a study that clarifies the connections between action and spontaneous potentials and glutamate-induced potentials. A different study by Paulmann et al. shows an indirect yet relevant contribution of the phloem's role as an electrical signaling network to determine the susceptibility of barley to viral infection. By altering the phloem's anatomy, the virus blocks the electrical message that would alert of its presence. Two research articles show new findings that underscore an unsuspected degree of sophistication of the plant electrical responses to environmental stimuli. On one hand, in their report of a new, protective role of local electrophysiological...
Simon Gilroy; Kazimierz Trebacz; Vicenta Salvador-Recatalà. Editorial: Inter-cellular Electrical Signals in Plant Adaptation and Communication. Frontiers in Plant Science 2018, 9, 643 .
AMA StyleSimon Gilroy, Kazimierz Trebacz, Vicenta Salvador-Recatalà. Editorial: Inter-cellular Electrical Signals in Plant Adaptation and Communication. Frontiers in Plant Science. 2018; 9 ():643.
Chicago/Turabian StyleSimon Gilroy; Kazimierz Trebacz; Vicenta Salvador-Recatalà. 2018. "Editorial: Inter-cellular Electrical Signals in Plant Adaptation and Communication." Frontiers in Plant Science 9, no. : 643.
The aim of the present study was to characterise bioelectrical changes in the membrane potential of Marchantia polymorpha gametophyte cells after light/dark transitions and to determine the role of the proton pump and energy status of the M. polymorpha cells in generation of these changes. Darkening caused persistent depolarisation of the resting potential (RP) and generation of short-lasting potential changes that were not uniform among different thalli. In some plants (18%), the changes evoked by darkening were typical action potentials (APsdark), whereas in 69% of the plants, the changes had a form of action potential-like responses (APsdark-like) consisting of a transient depolarisation followed by a plateau phase, whose magnitude and duration were inconstant. The illumination of the M. polymorpha always evoked action potentials (APslight) if the thallus was illuminated with light intensity of at least 120 µmol photons m−2 s−1 after 30-min darkening. To analyse the involvement of H+-ATPase in formation of the illumination/darkening-induced electrical responses in M. polymorpha, the proton pump regulators were used. The proton pump inhibitor (20 µM FCCP) significantly diminished the RP and inhibited dark-induced APdark and/or APdark-like responses and illumination-induced APslight. After application of DCMU (20 µM), the RP was strongly depolarised and no response to light/dark was observed. Fusicoccin (20 µM), i.e., an activator of the proton pump, strongly hyperpolarised the membrane potential and blocked dark-induced APdark/APdark-like responses and illumination-induced APslight.
Kamila Kupisz; Halina Dziubinska; Kazimierz Trebacz. Generation of action potential-type changes in response to darkening and illumination as indication of the plasma membrane proton pump status in Marchantia polymorpha. Acta Physiologiae Plantarum 2017, 39, 82 .
AMA StyleKamila Kupisz, Halina Dziubinska, Kazimierz Trebacz. Generation of action potential-type changes in response to darkening and illumination as indication of the plasma membrane proton pump status in Marchantia polymorpha. Acta Physiologiae Plantarum. 2017; 39 (3):82.
Chicago/Turabian StyleKamila Kupisz; Halina Dziubinska; Kazimierz Trebacz. 2017. "Generation of action potential-type changes in response to darkening and illumination as indication of the plasma membrane proton pump status in Marchantia polymorpha." Acta Physiologiae Plantarum 39, no. 3: 82.
Patch clamp recordings carried out in the inside-out configuration revealed activity of three kinds of channels: nonselective cation channels, small-conductance K+ channels, and large-conductance anion channels. The nonselective cation channels did not distinguish between Na+ and K+. The unitary conductance of these channels reached 28 pS in a symmetrical concentration of 200 mM NaCl. A lower value of this parameter was recorded for the small-conductance K+ channels and in a 50-fold gradient of K+ (200 mM/4 mM) it reached 8 pS. The high selectivity of these channels to potassium was confirmed by the reversal potential (-97 mV), whose value was close to the equilibrium potential for potassium (-100 mV). One of the features of the large-conductance anion channels was high conductance amounting to 493 pS in a symmetrical concentration of 200 mM NaCl. The channels exhibited three subconductance levels. Moreover, an increase in the open probability of the channels at voltages close to zero was observed. The anion selectivity of the channels was low, because the channels were permeable to both Cl- and gluconate – a large anion. Research on the calcium dependence revealed that internal calcium activates nonselective cation channels and small-conductance K+ channels, but not large-conductance anion channels.
M. Koselski; Anna Olszewska; A. Hordyjewska; T. Małecka-Massalska; Kazimierz Trebacz. Three Types of Ion Channels in the Cell Membrane of Mouse Fibroblasts. Physiological Research 2017, 63 -73.
AMA StyleM. Koselski, Anna Olszewska, A. Hordyjewska, T. Małecka-Massalska, Kazimierz Trebacz. Three Types of Ion Channels in the Cell Membrane of Mouse Fibroblasts. Physiological Research. 2017; ():63-73.
Chicago/Turabian StyleM. Koselski; Anna Olszewska; A. Hordyjewska; T. Małecka-Massalska; Kazimierz Trebacz. 2017. "Three Types of Ion Channels in the Cell Membrane of Mouse Fibroblasts." Physiological Research , no. : 63-73.
Potassium-permeable slow activating vacuolar channels (SV) and chloride-permeable channels in the vacuole of the liverwort Marchantia polymorpha were characterized in respect to calcium dependence, selectivity, and pharmacology. The patch-clamp method was used in the study of ion channel activity in the vacuoles from the liverwort Marchantia polymorpha. The whole-vacuole recordings allowed simultaneous observation of two types of currents-predominant slow activated currents recorded at positive voltages and fast activated currents recorded at negative voltages. Single-channel recordings carried out in the gradient of KCl indicated that slow activated currents were carried by potassium-permeable slowly activating vacuolar channels (SV) and fast activated currents-by chloride-permeable channels. Both types of the channels were dependent in an opposite way on calcium, since elimination of this ion from the cytoplasmic side caused inhibition of SV channels, but the open probability of chloride-permeable channels even increased. The dependence of the activity of both channels on different types of ion channel inhibitors was studied. SV channels exhibited different sensitivity to potassium channel inhibitors. These channels were insensitive to 3 mM Ba2+, but were blocked by 3 mM tetraethyl ammonium (TEA). Moreover, the activity of the channels was modified in a different way by calcium channel inhibitors. 200 µM Gd3+ was an effective blocker, but 50 µM ruthenium red evoked bursts of the channel activity resulting in an increase in the open probability. Different effectiveness of anion channel inhibitors was observed in chloride-permeable channels. After the application of 100 µM Zn2+, a decrease in the open probability was recorded but the channels were still active. 50 µM DIDS was more effective, as it completely blocked the channels.
Mateusz Koselski; Kazimierz Trebacz; Halina Dziubinska. Vacuolar ion channels in the liverwort Marchantia polymorpha: influence of ion channel inhibitors. Planta 2017, 245, 1049 -1060.
AMA StyleMateusz Koselski, Kazimierz Trebacz, Halina Dziubinska. Vacuolar ion channels in the liverwort Marchantia polymorpha: influence of ion channel inhibitors. Planta. 2017; 245 (5):1049-1060.
Chicago/Turabian StyleMateusz Koselski; Kazimierz Trebacz; Halina Dziubinska. 2017. "Vacuolar ion channels in the liverwort Marchantia polymorpha: influence of ion channel inhibitors." Planta 245, no. 5: 1049-1060.
Action potentials generated spontaneously (SAPs) and evoked by electrical stimulation (APs) in tomato plants (Solanum lycopersicum L.) cv. Micro-Tom ABA-deficient mutants (sitiens—MTsit) and its wild type (MTwt) were characterized by continuous monitoring of electrical activity for 66 h and by application of an electrical stimulation supplied extracellularly. MTsit generated SAPs which spread along the stem, including petioles and roots with an amplitude of 44.6 ± 4.4 mV, half-time (t½) of 33.1 ± 2.9 s and velocity of 5.4 ± 1.0 cm min−1. Amplitude and velocity were 43 and 108 % higher in MTsit than in MTwt, respectively. The largest number of SAPs was registered in the early morning in both genotypes. MTsit was less responsive to electrical stimuli. The excitation threshold and the refractory period were greater in MTsit than in MTwt. After current application, APs were generated in the MTwt with 21.2 ± 2.4 mV amplitude and propagated with 5.6 ± 0.5 cm min−1 velocity. Lower intensity stimuli did not trigger APs in these plants. In MTsit APs were measured with amplitude of 26.8 ± 4.8 mV and propagated with velocity of 8.5 ± 0.1 cm min−1.
F. C. O. Macedo; Halina Dziubinska; Kazimierz Trebacz; R. F. Oliveira; R. A. Moral. Action potentials in abscisic acid-deficient tomato mutant generated spontaneously and evoked by electrical stimulation. Acta Physiologiae Plantarum 2015, 37, 207 .
AMA StyleF. C. O. Macedo, Halina Dziubinska, Kazimierz Trebacz, R. F. Oliveira, R. A. Moral. Action potentials in abscisic acid-deficient tomato mutant generated spontaneously and evoked by electrical stimulation. Acta Physiologiae Plantarum. 2015; 37 (10):207.
Chicago/Turabian StyleF. C. O. Macedo; Halina Dziubinska; Kazimierz Trebacz; R. F. Oliveira; R. A. Moral. 2015. "Action potentials in abscisic acid-deficient tomato mutant generated spontaneously and evoked by electrical stimulation." Acta Physiologiae Plantarum 37, no. 10: 207.
Carotenoid pigments play numerous important physiological functions in human organism. Very special is a role of lutein and zeaxanthin in the retina of an eye and in particular in its central part, the macula lutea. In the retina, carotenoids can be directly present in the lipid phase of the membranes or remain bound to the protein–pigment complexes. In this work we address a problem of binding of carotenoids to proteins and possible role of such structures in pigment transport to lipid membranes. Interaction of three carotenoids, beta-carotene, lutein and zeaxanthin with two proteins: bovine serum albumin and glutathione S-transferase (GST) was investigated with application of molecular spectroscopy techniques: UV–Vis absorption, circular dichroism and Fourier transform infrared spectroscopy (FTIR). Interaction of pigment–protein complexes with model lipid bilayers formed with egg yolk phosphatidylcholine was investigated with application of FTIR, Raman imaging of liposomes and electrophysiological technique, in the planar lipid bilayer models. The results show that in all the cases of protein and pigment studied, carotenoids bind to protein and that the complexes formed can interact with membranes. This means that protein–carotenoid complexes are capable of playing physiological role in pigment transport to biomembranes.
Emilia Reszczynska; Renata Welc; Wojciech Grudzinski; Kazimierz Trebacz; Wieslaw I. Gruszecki. Carotenoid binding to proteins: Modeling pigment transport to lipid membranes. Archives of Biochemistry and Biophysics 2015, 584, 125 -133.
AMA StyleEmilia Reszczynska, Renata Welc, Wojciech Grudzinski, Kazimierz Trebacz, Wieslaw I. Gruszecki. Carotenoid binding to proteins: Modeling pigment transport to lipid membranes. Archives of Biochemistry and Biophysics. 2015; 584 ():125-133.
Chicago/Turabian StyleEmilia Reszczynska; Renata Welc; Wojciech Grudzinski; Kazimierz Trebacz; Wieslaw I. Gruszecki. 2015. "Carotenoid binding to proteins: Modeling pigment transport to lipid membranes." Archives of Biochemistry and Biophysics 584, no. : 125-133.
In this work, for the first time the activity of nitrate-permeable channels in the tonoplast of the moss Physcomitrella patens was recorded. The channels allowed nitrate flow in one direction-from the cytoplasm to the vacuole. Selectivity of nitrate over chloride of the channels was proved. The activity of the channels was dependent on cytoplasmic calcium, magnesium, and pH. A patch-clamp study carried out on the vacuolar membrane of the moss Physcomitrella patens has revealed that inhibition of cation-selective channels leads to disclosure of channels permeable to NO3 (-). These channels were inwardly rectifying and allowed anions to flow from the cytoplasm to the vacuole. After a decrease in the cytoplasmic NO3 (-) concentration, the current density recorded in the whole-vacuole configuration and amplitude of the currents flowing through single channels were reduced. Application of the NO3 (-) gradient caused a shift in the reversal potential (Erev) towards ENO3-, indicating NO3 (-) permeability. Research of the selectivity of the channels to Cl(-) and NO3 (-) was also done; it indicated that Cl(-) is less permeable than NO3 (-) (PNO3/PCl = 3.08). Measurements with different concentrations of cytoplasmic Ca(2+) and Mg(2+) revealed that the channel was activated by different concentrations of these ions-100 µM Ca(2+) and 10 mM Mg(2+). Calcium dependence of the channels was also modulated by a redox agent-DTT (dithiothreitol), which added on the cytoplasmic side, caused a reduction in the threshold of channel activation with cytoplasmic Ca(2+). The NO3 (-) permeable channel was also pH dependent. A decrease in the cytoplasmic pH reduced the open probability of the channel; in turn, an increase in the vacuolar pH did not decrease ion channel activity but lowered its conductance.
Mateusz Koselski; Halina Dziubinska; Aleksandra Seta-Koselska; Kazimierz Trebacz. A nitrate-permeable ion channel in the tonoplast of the moss Physcomitrella patens. Planta 2015, 241, 1207 -19.
AMA StyleMateusz Koselski, Halina Dziubinska, Aleksandra Seta-Koselska, Kazimierz Trebacz. A nitrate-permeable ion channel in the tonoplast of the moss Physcomitrella patens. Planta. 2015; 241 (5):1207-19.
Chicago/Turabian StyleMateusz Koselski; Halina Dziubinska; Aleksandra Seta-Koselska; Kazimierz Trebacz. 2015. "A nitrate-permeable ion channel in the tonoplast of the moss Physcomitrella patens." Planta 241, no. 5: 1207-19.
Amphotericin B (AmB) is an antifungal polyene for which the most accepted mode of action is formation of protein-like ion channels in the cell membrane. Patch-clamp research on Candida albicans protoplasts carried out in the outside-out configuration showed that application of 0.05 and 0.1 μM AmB caused a decrease in seal resistance. Such a phenomenon can be correlated with a decrease in membrane tightness. AmB applied at a 0.05 μM concentration also caused a decrease in the number of active TOK1 (two-pore outward rectifiers) potassium channels, but did not significantly change their open probability. The results indicate that in C. albicans protoplast AmB causes a decrease in cell membrane integrity by interaction with its lipid phase but not with ion channels. Fluorescence microscopy techniques showed that AmB treatment, in clinical concentrations, had no effect on the percentage of PI-positive protoplasts. AmB treatment in the concentrations tested did not cause a rapid reduction of the number of C. albicans protoplasts. However, there was a significant loss of replication competency and numerous morphological and physiological disorders, including cytoplasm shrinking, abnormal morphology of the nucleus and mitochondria, a sudden decrease in the MTT reduction level and oxidative stress. Our results show that the induction of yeast cell death by AmB, at therapeutic doses, is a multistage and long-term process involving multiple intracellular pathways.
Barbara Chudzik; Mateusz Koselski; Aleksandra Czuryło; Kazimierz Trebacz; Mariusz Gagoś. A new look at the antibiotic amphotericin B effect on Candida albicans plasma membrane permeability and cell viability functions. European Biophysics Journal 2015, 44, 77 -90.
AMA StyleBarbara Chudzik, Mateusz Koselski, Aleksandra Czuryło, Kazimierz Trebacz, Mariusz Gagoś. A new look at the antibiotic amphotericin B effect on Candida albicans plasma membrane permeability and cell viability functions. European Biophysics Journal. 2015; 44 (1):77-90.
Chicago/Turabian StyleBarbara Chudzik; Mateusz Koselski; Aleksandra Czuryło; Kazimierz Trebacz; Mariusz Gagoś. 2015. "A new look at the antibiotic amphotericin B effect on Candida albicans plasma membrane permeability and cell viability functions." European Biophysics Journal 44, no. 1: 77-90.
Our previous study has shown that the liverwort Conocephalum conicum generates action potentials (APs) in response to both temperature drop and menthol, which are also activators of the TRPM8 (transient receptor potential melastatin 8) receptor in animals. Not only similarities but also differences between electrical reactions to menthol and cooling observed in the liverwort aroused our interest in the action of menthol at the molecular level. Patch‐clamp investigations have shown that menthol causes a reduction of current flowing through slow vacuolar (SV) channels to 29 ± 10% of the initial value (n = 9); simultaneously, it does not influence magnitudes of currents passing through a single SV channel. This may point to an unspecific interaction between menthol and the lipid phase of the membrane. An influence of menthol on lipid organization in membranes was investigated in two‐component monomolecular layers formed with menthol and dipalmitoylphosphatidylcholine (DPPC) at the argon–water interface. Analyses of the mean molecular area parameters vs the molar fraction of the menthol component have shown over‐additivity (approximately 20 Å2) in the region of high molar fractions of menthol. Infrared absorption spectroscopy studies have shown that menthol, most probably, induces breaking of a hydrogen bond network formed by ester carbonyl groups and water bridges in the lipid membrane and binds to the polar head group region of DPPC. We conclude that the disruption in the lipid phase of the membrane influences ion channels and/or pumps and subsequently causes generation of APs in excitable plants such as C. conicum.
Kamila Kupisz; Kazimierz Trebacz; Wieslaw Gruszecki. Menthol-induced action potentials in Conocephalum conicum as a result of unspecific interactions between menthol and the lipid phase of the plasma membrane. Physiologia Plantarum 2014, 154, 349 -357.
AMA StyleKamila Kupisz, Kazimierz Trebacz, Wieslaw Gruszecki. Menthol-induced action potentials in Conocephalum conicum as a result of unspecific interactions between menthol and the lipid phase of the plasma membrane. Physiologia Plantarum. 2014; 154 (3):349-357.
Chicago/Turabian StyleKamila Kupisz; Kazimierz Trebacz; Wieslaw Gruszecki. 2014. "Menthol-induced action potentials in Conocephalum conicum as a result of unspecific interactions between menthol and the lipid phase of the plasma membrane." Physiologia Plantarum 154, no. 3: 349-357.
The effect of temperature upon the bioelectric potential across the plasma membrane in cells of tepals of Eranthis hyemalis (L.) Salisb. (Winter aconite) is described. Rapid warming of an intact tepal resulted in a transient small increase in the magnitude of transmembrane potential difference followed by a substantial long-lasting depolarization which is considered as an "anomalous" response. Upon rapid cooling the reverse response occurred: a small transient depolarization was followed by a substantial hyperpolarization (also an anomalous response). The anomalous responses were more pronounced in the epidermis on the abaxial side of the tepal than in that on the adaxial side, indicating an electrophysiological dorsiventrality of the tepals. The anomalous responses were much less apparent in cells of isolated tissues than in cells of intact tepals. This difference does not appear to result from wounding or bringing a tissue into direct contact with the external solution because in segments of tepals devoid of the abaxial epidermis only, the PD of the parenchyma behaved in a way similar to that of the intact tepals. It is suggested that the occurrence of the anomalous responses is modulated by the tissue stresses. The functional importance of the responses for thermonastic movements is discussed.
Zygmunt Hejnowicz; Kazimierz Trebacz; Andreas Sievers. Temperature-dependent changes of membrane potentials in cells of thermonastic tepals of Eranthis hyemalis (L.) Salisb. Acta Societatis Botanicorum Poloniae 2014, 63, 219 -227.
AMA StyleZygmunt Hejnowicz, Kazimierz Trebacz, Andreas Sievers. Temperature-dependent changes of membrane potentials in cells of thermonastic tepals of Eranthis hyemalis (L.) Salisb. Acta Societatis Botanicorum Poloniae. 2014; 63 (3-4):219-227.
Chicago/Turabian StyleZygmunt Hejnowicz; Kazimierz Trebacz; Andreas Sievers. 2014. "Temperature-dependent changes of membrane potentials in cells of thermonastic tepals of Eranthis hyemalis (L.) Salisb." Acta Societatis Botanicorum Poloniae 63, no. 3-4: 219-227.
Quercetin (3,3′,4′,5,7-pentahydroxyflavone) is claimed to exert many beneficial health effects. With application of 1H NMR (nuclear magnetic resonance) and FTIR (Fourier-transform infrared) techniques, quercetin interaction with liposomes formed with dipalmitoylphosphatidylcholine (DPPC) was analyzed. Patch-clamp technique was employed to study quercetin effects at single channel level of vacuolar membranes in the liverwort Conocephalum conicum. Light and electron microscopy were applied to study quercetin effects on human negroid cervix carcinoma cells (HeLa). Enzymatic measurements along with DPPH (1,1-diphenyl-2-picrylhydrazyl) bioassay were performed to investigate the influence of quercetin on antioxidant enzymes and reactive oxygen species (ROS) production. The inclusion of quercetin to the membrane exerted pronounced ordering effect on the motional freedom of lipids in the head group region as manifested by broadening of the 1H NMR spectral line representing the choline groups. FTIR analysis revealed quercetin incorporation into DPPC liposomes via hydrogen bonding between its own hydroxyl groups and lipid polar head groups in the C–O–P–O–C segment. Both, FTIR and NMR techniques indicated also quercetin spectral effects in the region corresponding to alkyl chains. Patch-clamp experiments showed that quercetin stabilizes tonoplast and promotes a close state of SV channels. Microscopic observations of HeLa cells revealed characteristic changes in ultrastructure and morphology of the examined cells in comparison to control cells. Pretreatment of HeLa cells with quercetin alleviated H2O2-induced cell injury by improving redox balance as indicated by the increase in glutathione content and SOD (superoxide dismutase) levels as well as by the decrease in ROS level. \In conclusion, the incorporation, distribution and the changes of biophysical properties of the membranes are very important for the effectiveness of phenolic compounds as antioxidant and anticancer factors
Bożena Pawlikowska-Pawlęga; Halina Dziubinska; Elżbieta Król; Kazimierz Trebacz; Anna Jarosz-Wilkolazka; Roman Paduch; Antoni Gawron; Wieslaw Gruszecki. Characteristics of quercetin interactions with liposomal and vacuolar membranes. Biochimica et Biophysica Acta (BBA) - Biomembranes 2014, 1838, 254 -265.
AMA StyleBożena Pawlikowska-Pawlęga, Halina Dziubinska, Elżbieta Król, Kazimierz Trebacz, Anna Jarosz-Wilkolazka, Roman Paduch, Antoni Gawron, Wieslaw Gruszecki. Characteristics of quercetin interactions with liposomal and vacuolar membranes. Biochimica et Biophysica Acta (BBA) - Biomembranes. 2014; 1838 (1):254-265.
Chicago/Turabian StyleBożena Pawlikowska-Pawlęga; Halina Dziubinska; Elżbieta Król; Kazimierz Trebacz; Anna Jarosz-Wilkolazka; Roman Paduch; Antoni Gawron; Wieslaw Gruszecki. 2014. "Characteristics of quercetin interactions with liposomal and vacuolar membranes." Biochimica et Biophysica Acta (BBA) - Biomembranes 1838, no. 1: 254-265.
Patch-clamp studies carried out on the tonoplast of the moss Physcomitrella patens point to existence of two types of cation-selective ion channels: slowly activated (SV channels), and fast-activated potassium-selective channels. Slowly and instantaneously saturating currents were observed in the whole-vacuole recordings made in the symmetrical KCl concentration and in the presence of Ca2+ on both sides of the tonoplast. The reversal potential obtained at the KCl gradient (10 mM on the cytoplasmic side and 100 mM in the vacuole lumen) was close to the reversal potential for K+ (E K), indicating K+ selectivity. Recordings in cytoplasm-out patches revealed two distinct channel populations differing in conductance: 91.6 ± 0.9 pS (n = 14) at −80 mV and 44.7 ± 0.7 pS (n = 14) at +80 mV. When NaCl was used instead of KCl, clear slow vacuolar SV channel activity was observed both in whole-vacuole and cytoplasm-out membrane patches. There were no instantaneously saturating currents, which points to impermeability of fast-activated potassium channels to Na+ and K+ selectivity. In the symmetrical concentration of NaCl on both sides of the tonoplast, currents have been measured exclusively at positive voltages indicating Na+ influx to the vacuole. Recordings with different concentrations of cytoplasmic and vacuolar Ca2+ revealed that SV channel activity was regulated by both cytoplasmic and vacuolar calcium. While cytoplasmic Ca2+ activated SV channels, vacuolar Ca2+ inhibited their activity. Dependence of fast-activated potassium channels on the cytoplasmic Ca2+ was also determined. These channels were active even without Ca2+ (2 mM EGTA in the cytosol and the vacuole lumen), although their open probability significantly increased at 0.1 μM Ca2+ on the cytoplasmic side. Apart from monovalent cations (K+ and Na+), SV channels were permeable to divalent cations (Ca2+ and Mg2+). Both monovalent and divalent cations passed through the channels in the same direction—from the cytoplasm to the vacuole. The identity of the vacuolar ion channels in Physcomitrella and ion channels already characterised in different plants is discussed.
Mateusz Koselski; Kazimierz Trebacz; Halina Dziubinska. Cation-permeable vacuolar ion channels in the moss Physcomitrella patens: a patch-clamp study. Planta 2013, 238, 357 -367.
AMA StyleMateusz Koselski, Kazimierz Trebacz, Halina Dziubinska. Cation-permeable vacuolar ion channels in the moss Physcomitrella patens: a patch-clamp study. Planta. 2013; 238 (2):357-367.
Chicago/Turabian StyleMateusz Koselski; Kazimierz Trebacz; Halina Dziubinska. 2013. "Cation-permeable vacuolar ion channels in the moss Physcomitrella patens: a patch-clamp study." Planta 238, no. 2: 357-367.
A plant is considered carnivorous if it receives any noticeable benefit from catching small animals. The morphological and physiological adaptations to carnivorous existence is most complex in plants, thanks to which carnivorous plants have been cited by Darwin as 'the most wonderful plants in the world'. When considering the range of these adaptations, one realizes that the carnivory is a result of a multitude of different features. This review discusses a selection of relevant articles, culled from a wide array of research topics on plant carnivory, and focuses in particular on physiological processes associated with active trapping and digestion of prey. Carnivory offers the plants special advantages in habitats where nutrient supply is scarce. Counterbalancing costs are the investments in synthesis and the maintenance of trapping organs and hydrolysing enzymes. With the progress in genetic, molecular and microscopic techniques, we are well on the way to a full appreciation of various aspects of plant carnivory. Sufficiently complex to be of scientific interest and finite enough to allow conclusive appraisal, carnivorous plants can be viewed as unique models for the examination of rapid organ movements, plant excitability, enzyme secretion, nutrient absorption, food-web relationships, phylogenetic and intergeneric relationships or structural and mineral investment in carnivory.
Elżbieta Król; Bartosz J. Płachno; Lubomír Adamec; Maria Stolarz; Halina Dziubinska; Kazimierz Trebacz. Quite a few reasons for calling carnivores ‘the most wonderful plants in the world’. Annals of Botany 2011, 109, 47 -64.
AMA StyleElżbieta Król, Bartosz J. Płachno, Lubomír Adamec, Maria Stolarz, Halina Dziubinska, Kazimierz Trebacz. Quite a few reasons for calling carnivores ‘the most wonderful plants in the world’. Annals of Botany. 2011; 109 (1):47-64.
Chicago/Turabian StyleElżbieta Król; Bartosz J. Płachno; Lubomír Adamec; Maria Stolarz; Halina Dziubinska; Kazimierz Trebacz. 2011. "Quite a few reasons for calling carnivores ‘the most wonderful plants in the world’." Annals of Botany 109, no. 1: 47-64.
The mechanism of cold perception by plants is still poorly understood. It was found that temperature drop evokes changes in the activity of ion pumps and channels, which leads to plasma membrane depolarization.1,2 The nature of the primary step of its action (alteration in membrane composition,3 transient influx of Ca2+ etc.,2) has not been elicited yet. Our electrophysiological experiments conducted on the liverwort Conocephalum conicum showed that its cells respond not only to sudden cooling4 but also to menthol, generating depolarization of the plasma membrane and action potentials (APs). Similar results are well documented in mammals; cold or “cooling compounds” including menthol cause activation of thermosenstitive channel TRPM8 permeable to Ca2+ and generation of AP series.5 TRP receptors are detected, among others, in green and brown algae. Possible existence of TRPM8-like channel-receptor in Conocephalum conicum is discussed here.
Kamila Kupisz; Kazimierz Trebacz. Electrophysiological approach to examine a putative cold- and menthol-sensitive channel in the liverwortConocephalum conicum. Plant Signaling & Behavior 2011, 6, 1002 -1003.
AMA StyleKamila Kupisz, Kazimierz Trebacz. Electrophysiological approach to examine a putative cold- and menthol-sensitive channel in the liverwortConocephalum conicum. Plant Signaling & Behavior. 2011; 6 (7):1002-1003.
Chicago/Turabian StyleKamila Kupisz; Kazimierz Trebacz. 2011. "Electrophysiological approach to examine a putative cold- and menthol-sensitive channel in the liverwortConocephalum conicum." Plant Signaling & Behavior 6, no. 7: 1002-1003.
In animals, cooling substances such as menthol are perceived as cold sensation because they bind to the same receptor TRPM8 (transient receptor potential melastatin) that activates upon temperature drops. We investigated the effect of menthol on the plant membrane potential to search for analogies between animal and plant perception systems. The study was conducted on the liverwort Conocephalum conicum- a non-vascular plant generating action potentials (APs) in response to different stimuli including cold. (+)Menthol, (-)menthol and (+/-)menthol induced one or more APs, depending on the concentration. In contrast to animal reactions to menthol, threshold concentrations of these isomers were the same (1 mM). The presence of menthol in medium shortened cold-induced APs, whereas low temperature prolonged the repolarization phase of AP evoked by menthol. Cells of C. conicum with anion and potassium channels blocked by anthracene-9-carboxylic acid (A9C) and tetraethylammonium chloride (TEACl) generate short spike-like voltage transients (VTs) in response to cold and light stimulation. Membrane potential changes evoked by menthol in A9C- and TEACl-treated plants differed significantly from VTs - lasted much longer and frequently occurred in series. 5 mM LaCl(3) , 1 mM EGTA (ethylene glycol-bis(2-aminoethyl ether)-N,N,N',N'-tetraacetic acid) (0 Ca(2+) ) but not 0.2 mM verapamil blocked the putative calcium component of AP induced by menthol. Similar inhibitory effect was observed after the application of proton pump inhibitors: 0.05 mM N,N-dicyclohexylcarbodiimide (DCCD), 0.05 mM diethylstilbestrol (DES) or 0.01 mM carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone (FCCP). Our results indicate that cold and menthol act independently, activating different membrane transporters in C. conicum cells.
Kamila Kupisz; Kazimierz Trebacz. Effect of cold and menthol on membrane potential in plants. Physiologia Plantarum 2011, 141, 352 -360.
AMA StyleKamila Kupisz, Kazimierz Trebacz. Effect of cold and menthol on membrane potential in plants. Physiologia Plantarum. 2011; 141 (4):352-360.
Chicago/Turabian StyleKamila Kupisz; Kazimierz Trebacz. 2011. "Effect of cold and menthol on membrane potential in plants." Physiologia Plantarum 141, no. 4: 352-360.
Apigenin, quercetin and genistein are members of the family of plant flavonoids suspected to prevent a number of human diseases, for instance cancer development. They display a number of activities, and part of their beneficial effects may be due to their affinity to cellular membranes. In this study, we used Conocephalum conicum, a well-elaborated model of liverworts. Intracellular microelectrode measurements were carried out to examine the effects of flavonoids in combination with neomycin on the resting and action potentials. Neomycin triggered gradual decline of action potential amplitudes through a membrane potential decrease (membrane potential became less negative) and a decrease of the action potential peak value. Additionally, duration of action potential amplitudes measured at half of the amplitude increased in neomycin-treated plants. However, the simultaneous use of quercetin or genistein (but not apigenin) with neomycin hindered neomycin-specific actions. Hence, the membrane resting potential and action potential amplitudes regained neomycin-free values. It may be concluded that application of at least some flavonoids (namely quercetin and genistein) exerts strong influence on electrical membrane responses in C. conicum.
Bożena Pawlikowska-Pawlęga; Elżbieta Król; Kazimierz Trebacz; Antoni Gawron. Quercetin and genistein hindering effect of neomycin action in the liverwort Conocephalum conicum. Acta Physiologiae Plantarum 2010, 33, 1335 -1344.
AMA StyleBożena Pawlikowska-Pawlęga, Elżbieta Król, Kazimierz Trebacz, Antoni Gawron. Quercetin and genistein hindering effect of neomycin action in the liverwort Conocephalum conicum. Acta Physiologiae Plantarum. 2010; 33 (4):1335-1344.
Chicago/Turabian StyleBożena Pawlikowska-Pawlęga; Elżbieta Król; Kazimierz Trebacz; Antoni Gawron. 2010. "Quercetin and genistein hindering effect of neomycin action in the liverwort Conocephalum conicum." Acta Physiologiae Plantarum 33, no. 4: 1335-1344.