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At least 19 recordsLinked to original sources

Different cytoplasmic calcium contents among three species of Characeae.

Internodal cells of three species of Characeae, Nitella flexilis, Nitella axilliformis and Chara corallina, were analyzed for the contents of Ca(2+ )and Mg(2+) in the cytoplasm. To avoid contamination of Ca(2+) from the cell wall and vacuole, the vacuolar sap was replaced with a sorbitol solution containing Sr(2+) by the vacuolar perfusion method after the cell had been treated with Sr(2+). No significant difference in the cytoplasmic content of Mg(2+) was found among three species of Characeae, but significant differences in the cytoplasmic content of Ca(2+) were observed among them. The cytoplasmic Ca(2+) content of N. flexilis was 2.0 times that of N. axilliformis and 3.3 times that of C. corallina. The cytoplasmic drop was furthermore separated into two fractions: a chloroplast-free fraction and a chloroplast fraction. In the chloroplast-free fraction the Ca(2+) content of N. flexilis was 2.3 times that of C. corallina and 2.0 times that of N. axilliformis, while the Mg(2+) content was the same among the three species. In the chloroplast fraction N. flexilis contained about seven times more Ca(2+) and about two times more Mg(2+) than C. corallina. The difference in the cytoplasmic Ca(2+ )content was discussed in relation to the difference in the capacity for the hydration-induced Ca(2+) release existing among the three species.

Calcium↗

Active movement of cardiac myosin on Characeae actin cables.

The active sliding of cardiac myosin on actin cables was studied using an in vitro movement assay. Cardiac myosin prepared from either adult rabbit or rat hearts was mixed with small latex beads to coat them. Actin cables were obtained from the internodal cells of green algae, Characeae. When the myosin-coated beads suspended in physiological buffer were introduced into the internodal cells, the myosin started to interact with the actin causing the beads to move. The sliding movement of the beads was observed under microscopy and the sliding velocity measured. The observed movement was smooth and the velocity was constant over a long distance. The movement was physiological in nature: a) it was ATP-dependent, but above a certain level of ATP, the velocity was constant; b) the velocity was maximum at pH 7.0, and decreased in both acidic and alkaline conditions. The average sliding velocity of cardiac myosin obtained from rabbit ventricles (0.31 +/- 0.11 micron/s) was slower than that from rat ventricles (1.04 +/- 0.26 micron/s) reflecting the lower ATPase activity of rabbit cardiac myosin. This assay system is considered to be a useful tool linking biochemistry and physiology at the molecular level.

Actins↗

Effective diameter and structural organization of reconstituted calcium channels from the Characeae algae Nitellopsis.

Using hydrophilic non-electrolytes, we determined the effective diameter of calcium channels from the Characeae alga Nitellopsis, reconstituted in a planar lipid membrane. It is suggested that a "single" reconstituted channel is a cluster consisting of several protochannels, in which the channel-forming molecules are stabilized by calcium ions. It is shown that the channels assembled in a cluster with the common conductivity of 350 pS (in 0.1 M KCl) have a funnel-like structure. The diameter of the smaller section is 0.96 +/- 0.20 nm, the diameter of the wider vestibule is 1.78 +/- 0.30 nm. Similarity of the structural organization of the channels investigated to the channels formed by annexin proteins is discussed.

Action Potentials↗

Studies on mechano-perception in the Characeae: transduction of pressure signals into electrical signals.

Mechano-perception by Chara cells was studied with an emphasis on the role of the nodal complex in transducing pressure signals into electrical signals. Three types of experimental material were used: (1) tandem internodal cells connected by a single layer of nodal cells; (2) single internodal cells, from which either apical or basal nodes were removed by ligation and cutting; (3) single internodes from which both nodes had been removed. Exposure to a hypertonic solution (sorbitol or sucrose) induced a depolarization at the node in 1 and 2. Depolarization did not occur at the ligated end of the cell in 2, or at all in 3. Addition of K+ increased the magnitude of the response, whilst it was significantly decreased by the divalent cations, Ca2+ and Mg2+. Electrical resistance decreased at the node during the depolarization, showing that a passive diffusion potential was responsible. I suggest that the change in the trans-nodal hydraulic pressure difference mechanically stretches the plasma membrane, and this induces the electrical depolarization.

Characeae↗

Ionic mechanism of mechano-perception in Characeae.

Characean internodal cells generate receptor potential in response to mechanical stimuli. We studied responses of internodal cells to a long-lasting stimulus and the results were as follows. (i) The cell generated receptor potential at the moment of both compression and decompression. (ii) The receptor potential (DeltaE (m)) was significantly larger at the moment of decompression than at compression. (iii) The longer the duration of the stimulus, the larger was the magnitude of DeltaE (m) at the moment of decompression. (iv) Aequorin studies revealed that the increase in [Ca(2+)](c) (Delta[Ca(2+)](c)) took place at the moment of both compression and decompression. (v) The amplitude of Delta[Ca(2+)](c) was larger at the moment of decompression than at compression, as was the case for DeltaE (m). It was suggested that the amplitude of the receptor potential had a tight correlation with the degree of membrane deformation. We discussed the ionic mechanism of mechano-perception under a long-lasting stimulus in relation to mechanosensitive activation of Ca(2+) channels at the plasma membrane.

Calcium↗

Kinetics studies of ultraweak luminescence induced by ascorbic acid in Characeae cells and their structures.

Native non-stressed Nitellopsis obtusa cells emit low-intensity (10-10(4) photons s(-1) cm(-2)) electromagnetic radiation within the visible range. The intensity of emission by these cells is several times higher than the intensity of the environmental medium. This luminescence appears as a long-lived (observable not for hours but for a few days) spontaneous light emission, sensitized by chlorophyll. Cells exposed to ascorbic acid action emit ultraweak radiation, dependent on reagent concentrations. Ultraweak luminescence (UL) intensity increases with rise in concentration, reaching maximum for higher concentrations faster and also changing the kinetics of UL decay. Studies of UL kinetics for different substructures and fractions, as well as for intact cells, allow us to consider single- and double-exponential functions as decay curves, which could suggest two ways of ascorbic acid action in the cells. In the spectral distribution, the bands of chlorophyll, dimoles of singlet oxygen and NADH, FMN and Q(10), which are electron carriers of the respiration chain, can be taken into consideration. Ascorbic acid action shows an increase in the red part of spectrum, except 627 nm, and in these bands of emission, which can be associated with the electron carriers of the respiration chain and lipid peroxidation.

Ascorbic Acid↗

D2O-induced ion channel activation in Characeae at low ionic strength.

Effects of D2O were studied on internodal cells of the freshwater alga Nitellopsis obtusa under plasmalemma perfusion (tonoplast-free cells) with voltage clamp, and on Ca2+ channels isolated from the alga and reconstituted in bilayer lipid membranes (BLM). External application of artificial pond water (APW) with D2O as the solvent to the perfused plasmalemma preparation led to an abrupt drop of membrane resistance (Rm = 0.12 +/- 0.03 k omega.cm2), thus preventing further voltage clamping. APW with 25% D2O caused a two-step reduction of Rm: first, down to 2.0 +/- 0.8 k omega.cm2, and then further to 200 omega.cm2, in 2 min. It was shown that in the first stage, Ca2+ channels are activated, and then, Ca2+ ions entering through them activate the Cl- channels. The Ca2+ channels are activated irreversibly. If 100 mM CsCl was substituted for 200 mM sucrose (introduced for iso-osmoticity), no effect of D2O on Rm was observed. Intracellular H2O/D2O substitution also did not change Rm. In experiments on single Ca2+ channels in BLM H2O/D2O substitution in a solution containing 100 mM KCl (trans side) produced no effect on channel activity, while in 10 mM KCl, at negative voltage, the open channel probability sharply increased. This effect was irreversible. The single channel conductance was not altered after the H2O/D2O substitution. The discussion of the possible mechanism of D2O action on Ca2+ and Cl- channels was based on an osmotic-like stress effect and the phenomenon of higher D-bond energy compared to the H-bond.

Calcium Channels↗

Blockers of Ca2+ channels in the plasmalemma of perfused Characeae cells.

Ionic currents in the plasmalemma of perfused Nitella syncarpa cells identified as currents through Ca2+ channels were registered for the first time. The effect of 1,4-dihydropyridine derivatives (nifedipine, nitrendipine, riodipine) and phenylalkylamines (verapamil, D600) as well as the agonist CGP-28392 on the Ca2+ channels in the plasmalemma of perfused cells of Nitellopsis obtusa and Nitella syncarpa have been studied. A blocking effect of 1,4-dihydropyridine derivatives and phenylalkylamines on the plasmalemma Ca2+ channels has been detected. Phenylalkylamines have been found to block both inward and outward Ca2+ currents. The activating effect of the agonist CGP-28392 on the Ca2+ channels of plasmalemma has been shown.

Calcium Channel Agonists↗

Accelerated sliding of pollen tube organelles along Characeae actin bundles regulated by Ca2+.

Pollen tubes show active cytoplasmic streaming. We isolated organelles from pollen tubes and tested their ability to slide along actin bundles in characean cell models. Here, we show that sliding of organelles was ATP-dependent and that motility was lost after N-ethylmaleimide or heat treatment of organelles. On the other hand, cytoplasmic streaming in pollen tube was inhibited by either N-ethylmaleimide or heat treatment. These results strongly indicate that cytoplasmic streaming in pollen tubes is supported by the "actomyosin"-ATP system. The velocity of organelle movement along characean actin bundles was much higher than that of the native streaming in pollen tubes. We suggested that pollen tube "myosin" has a capacity to move at a velocity of the same order of magnitude as that of characean myosin. Moreover, the motility was high at Ca2+ concentrations lower than 0.18 microM (pCa 6.8) but was inhibited at concentration higher than 4.5 microM (pCa 5.4). In conclusion, cytoplasmic streaming in pollen tubes is suggested to be regulated by Ca2+ through "myosin" inactivation.

Actins↗

ATP-Regulated Ion Channels in the Plasma Membrane of a Characeae Alga, Nitellopsis obtusa.

Using the patch-clamp technique, we recorded single-channel currents across the excised patch of the plasma membrane of Nitellopsis. Both K(+) and Na(+) can pass this channel, but currents were not carried by Cl(-). Upon the addition of ATP or AMP to the cytoplasmic side, the frequency of channel opening decreased. This is the first report on an ATP-regulated channel in plant cells.

Journal Article↗

The effect of the external medium on the gravity-induced polarity of cytoplasmic streaming in Chara corallina (Characeae).

Gravity induces a polarity of cytoplasmic streaming in vertical internodal cells of Chara such that the downwardly directed stream moves faster than the upwardly directed stream. In order to determine whether the statolith theory (in which intracellular sedimenting particles are responsible for gravity sensing) or the gravitational pressure theory (in which the entire protoplast acts as the gravity sensor) best explain the gravity response in Chara internodal cells, we controlled the physical properties of the external medium, including density and osmolarity, with impermeant solutes and examined the effect on the polarity of cytoplasmic streaming. As the density of the external medium is increased, the polarity of cytoplasmic streaming decreases and finally disappears when the density of the external medium is equal to that of the cell (1015 kg/m3). A further increase in the density of the external medium causes a reversal of the gravity response. These results are consistent with the gravitational pressure theory of gravity sensing since the buoyancy of the protoplast is dependent on the difference between the density of the protoplast and the external medium, and are inconsistent with the statolith theory since the buoyancy of intracellular particles are unaffected by changes in the external medium.

Biophysical Phenomena↗