Single Ion Channel Current Data from the Plasmalemma of Cytoplasmic Fragments of the Green Alga, Hydrodictyon africanum

1995 ◽  
Vol 22 (4) ◽  
pp. 571
Author(s):  
N Findlay ◽  
GP Findlay
Keyword(s):  
Ion Channels ◽  
Higher Plants ◽  
Membrane Surface ◽  
Plant Cells ◽  
Outward Current ◽  
Current Data ◽  
Patch Clamping ◽  
Ion Specificity ◽  
Channel Current ◽  
Gain Access

To measure, by patch clamping, electric current through ion channels in the plasmalemma of plant cells, access to the membrane surface is required. In higher plants, this access is gained by the preparation of protoplasts by enzymic methods. In plants such as the freshwater alga Hydrodictyon africanum, which have large cells, the preparation of protoplasts by enzymic methods is not possible. In this paper we describe a non-enzymic method for gaining access to the plasmalemma of Hydrodictyon. Our initial attempts to gain access to the plasmalemma of these cells by first plasmolysing the cells, and then cutting a window in the wall, so exposing the plasmalemma, were not successful. It was possible, however, after drying the cells in air, to cut them open and obtain fragments of the cytoplasm which maintain their original curvature. Standard patch clamping methods were then used to measure currents through single ion channels in the membrane, presumed to be the plasmalemma, bounding the outer surface of the fragments, although the success rate was low. Two types of channels were observed: (a) a multistate channel whose ion specificity was not established, and which in attached patches behaved as an outward current rectifier, and in a detached patch, as both inward and outward rectifier; and (b) a Cl- channel.

scholarly journals Stochastic Spatial Heterogeneity in Activities of H+-ATP-ases in Electrically Connected Plant Cells Decreases Threshold for Cooling-Induced Electrical Responses

2021 ◽  
Vol 22 (15) ◽  
pp. 8254
Author(s):  
Ekaterina Sukhova ◽  
Daria Ratnitsyna ◽  
Vladimir Sukhov
Keyword(s):  
Spatial Heterogeneity ◽  
Action Potentials ◽  
Higher Plants ◽  
Electrical Response ◽  
Plant Cells ◽  
Excitable Cells ◽  
Molecular Transporters ◽  
Positive Shift ◽  
Stochastic Heterogeneity ◽  
Electrical Responses

H+-ATP-ases, which support proton efflux through the plasma membrane, are key molecular transporters for electrogenesis in cells of higher plants. Initial activities of the transporters can influence the thresholds of generation of electrical responses induced by stressors and modify other parameters of these responses. Previously, it was theoretically shown that the stochastic heterogeneity of individual cell thresholds for electrical responses in a system of electrically connected neuronal cells can decrease the total threshold of the system (“diversity-induced resonance”, DIR). In the current work, we tested a hypothesis about decreasing the thresholds of generation of cooling-induced electrical responses in a system of electrically connected plant cells with increasing stochastic spatial heterogeny in the initial activities of H+-ATP-ases in these cells. A two-dimensional model of the system of electrically connected excitable cells (simple imitation of plant leaf), which was based on a model previously developed in our works, was used for the present investigation. Simulation showed that increasing dispersion in the distribution of initial activities of H+-ATP-ases between cells decreased the thresholds of generation of cooling-induced electrical responses. In addition, the increasing weakly influenced the amplitudes of electrical responses. Additional analysis showed two different mechanisms of the revealed effect. The increasing spatial heterogeneity in activities of H+-ATP-ases induced a weak positive shift of the membrane potential at rest. The shift decreased the threshold of electrical response generation. However, the decreased threshold induced by increasing the H+-ATP-ase activity heterogeneity was also observed after the elimination of the positive shift. The result showed that the “DIR-like” mechanism also participated in the revealed effect. Finally, we showed that the standard deviation of the membrane potentials before the induction of action potentials could be used for the estimation of thresholds of cooling-induced plant electrical responses. Thus, spatial heterogeneity in the initial activities of H+-ATP-ases can be a new regulatory mechanism influencing the generation of electrical responses in plants under actions of stressors.

Calcium/Calmodulin Affects Microtubule Stability in Lysed Protoplasts

1991 ◽  
Vol 100 (2) ◽  
pp. 311-317
Author(s):  
RICHARD J. CYR
Keyword(s):  
Cell Cycle ◽  
Dynamic Behavior ◽  
Growth And Development ◽  
Higher Plants ◽  
Plant Cells ◽  
Microtubule Stability ◽  
A Cell ◽  
Cycle Dependent ◽  
Carrot Protoplasts

Microtubules (Mts) are found in four distinct arrays appearing sequentially in a cell-cycle-dependent fashion within the cells of higher plants. Additionally, the cortical Mts of non-cycling cells are spatially altered in a variety of differentiated states. Information regarding the molecular details underlying these Mt-reorientation events in plant cells is scarce. Moreover, it is unclear how cytoskeletal behavior integrates with the myriad of other cellular activities that are altered concomitantly in both differentiating and cycling cells. Data are presented herein to indicate that calcium, in the form of a Ca2+/calmodulin complex, can alter the behavior of Mts in lysed carrot protoplasts. Mechanistically, we show that Ca2+/calmodulin most likely interacts with Mts via associations with microtubule associated pro- teins (MAPS). These results are discussed with reference to how Ca2+ may alter the dynamic behavior of Mts during growth and development.

Ion Channels in the Plasma Membrane of Plant Cells

1992 ◽  
pp. 225-236
Author(s):  
B. R. Terry ◽  
S. D. Tyerman ◽  
G. P. Findlay
Keyword(s):  
Plasma Membrane ◽  
Ion Channels ◽  
Plant Cells

scholarly journals Cytokinesis in tobacco BY-2 and root tip cells: a new model of cell plate formation in higher plants.

1995 ◽  
Vol 130 (6) ◽  
pp. 1345-1357 ◽  
Author(s):  
A L Samuels ◽  
T H Giddings ◽  
L A Staehelin
Keyword(s):  
Higher Plants ◽  
Membrane Surface ◽  
Cell Plate ◽  
Root Tip ◽  
Cellulose Synthesis ◽  
Parent Cell ◽  
Cell Periphery ◽  
New Model ◽  
Cell Plate Formation ◽  
Plate Formation

Cell plate formation in tobacco root tips and synchronized dividing suspension cultured tobacco BY-2 cells was examined using cryofixation and immunocytochemical methods. Due to the much improved preservation of the cells, many new structural intermediates have been resolved, which has led to a new model of cell plate formation in higher plants. Our electron micrographs demonstrate that cell plate formation consists of the following stages: (1) the arrival of Golgi-derived vesicles in the equatorial plane, (2) the formation of thin (20 +/- 6 nm) tubes that grow out of individual vesicles and fuse with others giving rise to a continuous, interwoven, tubulo-vesicular network, (3) the consolidation of the tubulo-vesicular network into an interwoven smooth tubular network rich in callose and then into a fenestrated plate-like structure, (4) the formation of hundreds of finger-like projections at the margins of the cell plate that fuse with the parent cell membrane, and (5) cell plate maturation that includes closing of the plate fenestrae and cellulose synthesis. Although this is a temporal chain of events, a developing cell plate may be simultaneously involved in all of these stages because cell plate formation starts in the cell center and then progresses centrifugally towards the cell periphery. The "leading edge" of the expanding cell plate is associated with the phragmoplast microtubule domain that becomes concentrically displaced during this process. Thus, cell plate formation can be summarized into two phases: first the formation of a membrane network in association with the phragmoplast microtubule domain; second, cell wall assembly within this network after displacement of the microtubules. The phragmoplast microtubules end in a filamentous matrix that encompasses the delicate tubulo-vesicular networks but not the tubular networks and fenestrated plates. Clathrin-coated buds/vesicles and multivesicular bodies are also typical features of the network stages of cell plate formation, suggesting that excess membrane material may be recycled in a selective manner. Immunolabeling data indicate that callose is the predominant lumenal component of forming cell plates and that it forms a coat-like structure on the membrane surface. We postulate that callose both helps to mechanically stabilize the early delicate membrane networks of forming cell plates, and to create a spreading force that widens the tubules and converts them into plate-like structures. Cellulose is first detected in the late smooth tubular network stage and its appearance seems to coincide with the flattening and stiffening of the cell plate.

scholarly journals Peripheral Mechanobiology of Touch—Studies on Vertebrate Cutaneous Sensory Corpuscles

2020 ◽  
Vol 21 (17) ◽  
pp. 6221 ◽  
Author(s):  
Ramón Cobo ◽  
Jorge García-Piqueras ◽  
Yolanda García-Mesa ◽  
Jorge Feito ◽  
Olivia García-Suárez ◽  
...  
Keyword(s):  
Ion Channels ◽  
Transient Receptor Potential ◽  
Receptor Potential ◽  
Transient Receptor Potential Channel ◽  
Mechanical Stimulus ◽  
Nerve Fibers ◽  
Current Data ◽  
Acid Sensing Ion Channels ◽  
Low Threshold ◽  
Transient Receptor

The vertebrate skin contains sensory corpuscles that are receptors for different qualities of mechanosensitivity like light brush, touch, pressure, stretch or vibration. These specialized sensory organs are linked anatomically and functionally to mechanosensory neurons, which function as low-threshold mechanoreceptors connected to peripheral skin through Aβ nerve fibers. Furthermore, low-threshold mechanoreceptors associated with Aδ and C nerve fibers have been identified in hairy skin. The process of mechanotransduction requires the conversion of a mechanical stimulus into electrical signals (action potentials) through the activation of mechanosensible ion channels present both in the axon and the periaxonal cells of sensory corpuscles (i.e., Schwann-, endoneurial- and perineurial-related cells). Most of those putative ion channels belong to the degenerin/epithelial sodium channel (especially the family of acid-sensing ion channels), the transient receptor potential channel superfamilies, and the Piezo family. This review updates the current data about the occurrence and distribution of putative mechanosensitive ion channels in cutaneous mechanoreceptors including primary sensory neurons and sensory corpuscles.

Voltage- and transmitter-gated currents in isolated rod bipolar cells of rat retina

1990 ◽  
Vol 63 (4) ◽  
pp. 860-876 ◽  
Author(s):  
A. Karschin ◽  
H. Wassle
Keyword(s):  
Ion Channels ◽  
Outward Current ◽  
Amacrine Cells ◽  
Bipolar Cells ◽  
Similar Distribution ◽  
Isolated Cells ◽  
Membrane Hyperpolarization ◽  
Rat Retina ◽  
Inward Current ◽  
Rod Bipolar Cells

1. Bipolar cells were isolated from adult rat retinas after enzymatic and mechanical treatment. The cells could be unequivocally identified from their morphology because of high retention of their axon and dendritic processes after isolation. 2. Protein kinase C (PKC) immunoreactivity performed on sections of the rat retina labeled rod bipolar cells and a few amacrine cells. Virtually all bipolar cells in the dissociates expressed PKC immunoreactivity and were, therefore, rod bipolar cells. 3. Rod bipolar cells were examined with the tight-seal whole-cell and excised-patch recording techniques. Resting potentials of the isolated cells recorded under current-clamp conditions showed a broad unimodal distribution around -37 mV. 4. Membrane depolarization from a holding potential of -90 mV resulted in an outward current. A fast sodium inward current was not observed. Membrane hyperpolarization from a holding potential of -40 mV activated an inwardly rectifying current. 5. gamma-Aminobutyric acid (GABA) and glycine, the putative retinal neurotransmitters that mediate the bipolar cells' receptive field surround in vivo, activated chloride conductances in almost all isolated bipolar cells. GABA- and glycine-evoked currents were both desensitizing and could be antagonized by the classical blockers bicuculline, picrotoxin, and strychnine, respectively. 6. Pressure application of the drugs from fine microcapillaries to various parts of the isolated cells suggests a high GABA sensitivity at the axonal endings compared with either the somatic or dendritic region. A similar distribution was not found for glycine. On the contrary, glycine-induced single-channel events with main conductances of 52 and 34 pS were recorded from membrane patches excised from the cells' somata. 7. Conductances induced by glutamate and several excitatory amino acid agonists were observed in a number of the cells. Application of the glutamate agonist 2-amino-4-phosphonobutyric acid (APB) induced an inward current at negative holding potentials associated with the opening of ion channels. In only 5 of 93 cells, APB closed ion channels, leading to a decrease in membrane conductance.

Calreticulin, a multifunctional Ca2+ binding chaperone of the endoplasmic reticulum

1998 ◽  
Vol 76 (5) ◽  
pp. 779-785 ◽  
Author(s):  
Marek Michalak ◽  
Paola Mariani ◽  
Michal Opas
Keyword(s):  
Gene Expression ◽  
Endoplasmic Reticulum ◽  
Posttranslational Modification ◽  
Storage Capacity ◽  
Higher Plants ◽  
Plant Cells ◽  
Steroid Sensitive ◽  
Endoplasmic Reticulum Chaperone ◽  
Precise Role ◽  
Cellular Ca

Calreticulin is a ubiquitous endoplasmic reticulum Ca2+ binding chaperone. The protein has been implicated in a variety of diverse functions. Calreticulin is a lectin-like chaperone and, together with calnexin, it plays an important role in quality control during protein synthesis, folding, and posttranslational modification. Calreticulin binds Ca2+ and affects cellular Ca2+ homeostasis. The protein increases the Ca2+ storage capacity of the endoplasmic reticulum and modulates the function of endoplasmic reticulum Ca2+-ATPase. Calreticulin also plays a role in the control of cell adhesion and steroid-sensitive gene expression. Recently, the protein has been identified and characterized in higher plants but its precise role in plant cells awaits further investigation.Key words: calreticulin, endoplasmic reticulum, chaperone, Ca2+ binding protein.

scholarly journals The thermal limits to life on Earth

2014 ◽  
Vol 13 (2) ◽  
pp. 141-154 ◽  
Author(s):  
Andrew Clarke
Keyword(s):  
Life Cycle ◽  
Higher Plants ◽  
Temperature Limit ◽  
Current Data ◽  
High Temperature Limit ◽  
Thermal Limits ◽  
Resource Limited ◽  
Almost Everywhere ◽  
Domains Of Life ◽  
Living Organisms

AbstractLiving organisms on Earth are characterized by three necessary features: a set of internal instructions encoded in DNA (software), a suite of proteins and associated macromolecules providing a boundary and internal structure (hardware), and a flux of energy. In addition, they replicate themselves through reproduction, a process that renders evolutionary change inevitable in a resource-limited world. Temperature has a profound effect on all of these features, and yet life is sufficiently adaptable to be found almost everywhere water is liquid. The thermal limits to survival are well documented for many types of organisms, but the thermal limits to completion of the life cycle are much more difficult to establish, especially for organisms that inhabit thermally variable environments. Current data suggest that the thermal limits to completion of the life cycle differ between the three major domains of life, bacteria, archaea and eukaryotes. At the very highest temperatures only archaea are found with the current high-temperature limit for growth being 122 °C. Bacteria can grow up to 100 °C, but no eukaryote appears to be able to complete its life cycle above ∼60 °C and most not above 40 °C. The lower thermal limit for growth in bacteria, archaea, unicellular eukaryotes where ice is present appears to be set by vitrification of the cell interior, and lies at ∼−20 °C. Lichens appear to be able to grow down to ∼−10 °C. Higher plants and invertebrates living at high latitudes can survive down to ∼−70 °C, but the lower limit for completion of the life cycle in multicellular organisms appears to be ∼−2 °C.

scholarly journals Development of an automated system to measure ion channel currents using a surface-modified gold probe

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Minako Hirano ◽  
Masahisa Tomita ◽  
Chikako Takahashi ◽  
Nobuyuki Kawashima ◽  
Toru Ide
Keyword(s):  
Ion Channels ◽  
Ion Channel ◽  
Lipid Bilayer ◽  
Measurement System ◽  
Automated System ◽  
Channel Activity ◽  
Channel Current ◽  
Gold Probe ◽  
Measurement Efficiency ◽  
Channel Currents

AbstractArtificial lipid bilayer single-channel recording technique has been employed to determine the biophysical and pharmacological properties of various ion channels. However, its measurement efficiency is very low, as it requires two time-consuming processes: preparation of lipid bilayer membranes and incorporation of ion channels into the membranes. In order to address these problems, we previously developed a technique based on hydrophilically modified gold probes on which are immobilized ion channels that can be promptly incorporated into the bilayer membrane at the same time as the membrane is formed on the probes’ hydrophilic area. Here, we improved further this technique by optimizing the gold probe and developed an automated channel current measurement system. We found that use of probes with rounded tips enhanced the efficiency of channel current measurements, and introducing a hydrophobic area on the probe surface, beside the hydrophilic one, further increased measurement efficiency by boosting membrane stability. Moreover, we developed an automated measurement system using the optimized probes; it enabled us to automatically measure channel currents and analyze the effects of a blocker on channel activity. Our study will contribute to the development of high-throughput devices to identify drug candidates affecting ion channel activity.

Sign in / Sign up

Export Citation Format

Share Document