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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.
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.
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.
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.
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.
Intracellular microelectrode measurements revealed that a resting potential (RP), an action potential (AP) and a calcium component of AP (named voltage transient, VT) can be influenced by glutamic acid (Glu) and aminoacetic acid (glycine, Gly) in the liverwort Conocephalum conicum. In the continuous presence of 5 mM Glu or 5 mM Gly, the RP hyperpolarized constantly and the plants became desensitized to the excitatory amino acids (Glu or Gly). Under such circumstances, the amplitudes of APs evoked by stimuli other than Glu or Gly grew, as did their calcium components (VTs). The sudden application of 1–15 mM Glu or Gly to a thallus not yet desensitized resulted in an excitation, i.e. a single AP or AP series. Aspartate (Asp) could not substitute for Glu in any way. Simultaneous action of both amino acids acted synergically to trigger APs. The same phenomenon was observed when glycine solution was enriched with N-methyl-d-aspartic acid (NMDA). Gly-induced APs were totally hindered by 1 mM d-amino-5-phosphonopentanoic acid (AP5) – an inhibitor of ionotropic glutamate receptors of the NMDA kind. Glu-induced APs could be totally suppressed by 1 mM AP5 as well as by 1 mM 6,7-dinitroquinoxaline-2,3-dione (DNQX) – an inhibitor of AMPA/KA receptors. DNQX also completely blocked the calcium component of Glu-evoked APs. After DNQX treatment, the only response to Glu was a membrane potential hyperpolarization (like the Glu response in a desensitized plant). It was concluded that the Glu-induced depolarization and hyperpolarization are separate phenomena. The stimulatory effects of both Glu and Gly on liverwort excitability may be the consequences of an activation of a variety of ionotropic Glu receptor subtypes.
Elzbieta Krol; Halina Dziubinska; Kazimierz Trebacz; Mateusz Koselski; Maria Stolarz. The influence of glutamic and aminoacetic acids on the excitability of the liverwort Conocephalum conicum. Journal of Plant Physiology 2007, 164, 773 -784.
AMA StyleElzbieta Krol, Halina Dziubinska, Kazimierz Trebacz, Mateusz Koselski, Maria Stolarz. The influence of glutamic and aminoacetic acids on the excitability of the liverwort Conocephalum conicum. Journal of Plant Physiology. 2007; 164 (6):773-784.
Chicago/Turabian StyleElzbieta Krol; Halina Dziubinska; Kazimierz Trebacz; Mateusz Koselski; Maria Stolarz. 2007. "The influence of glutamic and aminoacetic acids on the excitability of the liverwort Conocephalum conicum." Journal of Plant Physiology 164, no. 6: 773-784.