Depolarization of cell membranes in leaves of Lycopersicon by extract containing Ricca's factor

1977 ◽  
Vol 55 (5) ◽  
pp. 511-519 ◽  
Author(s):  
John M. Cheeseman ◽  
Barbara G. Pickard
Keyword(s):  
Membrane Potential ◽  
Maximum Rate ◽  
Electrical Coupling ◽  
Leaf Tissue ◽  
Saturation Level ◽  
Free Medium ◽  
Fresh Leaf ◽  
Stable Level ◽  
Shoot Tissue ◽  
Electrophysiological Studies

Extract containing Ricca's factor depolarizes the membrane potential of at least three types of cells in Lycopersicon leaves : mesophyll, midrib parenchyma, and midrib epidermis. The depolarization has been studied in some detail for the epidermal cells, in which depolarization appears to begin without a lag and is completed within 60–90 s. The maximum rate of depolarization is typically about 3 mV s−1. No changes in resistivity, capacivity, or intercellular electrical coupling have been detected during the depolarization.Extract from 0.5 mg fresh leaf tissue in 1 ml of water at pH 6.6 causes threshold depolarization in many experiments, and a concentration only 40 times greater is usually saturating. Raising the pH increases the concentration of factor required for saturation.With subsaturating concentrations of factor, the potential recovers somewhat after depolarization, and when factor-free medium is washed over the tissue the potential depolarizes briefly before returning to its baseline value. With saturating concentrations of factor, the potential depolarizes to an essentially stable level and no transient depolarization occurs when the factor is washed out.The potential remaining after application of a saturating concentration of factor is independent of the initial baseline potential but depends on the concentration of K+ in the equilibration medium and in the extract. The saturation level of depolarization is in the range of the Nernstian potential for K+, but whether it is precisely equal to the Nernstian potential for K+ has not been established.Evidently, the occurrence and influence of Ricca's factor should be taken into account in all electrophysiological studies of shoot tissue since the factor appears to be released whenever cells are wounded and may be released during other kinds of stress as well.

scholarly journals Ion Absorption by Shoot Tissue: Technique and First Findings with Excised Leaf Tissue of Corn

1964 ◽  
Vol 39 (3) ◽  
pp. 338-341 ◽  
Author(s):  
Richard C. Smith ◽  
Emanuel Epstein
Keyword(s):  
Leaf Tissue ◽  
Shoot Tissue ◽  
Ion Absorption

Phosphorus compounds measured in a rapid and simple leaf test for the assessment of the phosphorus status of subterranean clover

1984 ◽  
Vol 24 (125) ◽  
pp. 213 ◽  
Author(s):  
GCJ Irving ◽  
D Bouma
Keyword(s):  
Leaf Tissue ◽  
Rapid Test ◽  
Subterranean Clover ◽  
Exchange Chromatography ◽  
Phosphorus Compounds ◽  
Fresh Leaf ◽  
Organic Phosphates ◽  
Phosphorus Status ◽  
Colour Development ◽  
Hydrolysis Of

Experiments were done to determine what proportion of the phosphate extracted from fresh leaf tissue by five drops of 10 N H2SO4 represents inorganic tissue phosphate, and to what extent hydrolysis of organic phosphates during and after the extraction, and during the development of the blue phosphomolybdate complex, could contribute to the values obtained. The extraction is the basis of a simple and rapid test for the assessment of the phosphorus status of subterranean clover (Bouma and Dowling 1982). Extraction of leaf tissue of subterranean clover and sunflower with 0.2 M HClO4 at O�C, which was shown to extract inorganic leaf phosphorus without causing significant hydrolysis of organic phosphates, gave values not significantly different from those in H2SO4 extracts. The rate of hydrolysis of endogenous organic phosphates in tissue, extracted and left at room temperature for periods of up to 40 min. after adding H2SO4, did not differ significantly from zero. Errors due to hydrolysis during the 30 min. previously recommended for colour development are reduced to negligible proportions by reducing the time for colour development to 10 min. and by adding citric acid at this point. Anion-exchange chromatography of 10 N H2SO4 and 0.2 M HClO4 extracts confirmed the similarity of their composition and provided estimates of the various phosphate compounds present. The extraction of fresh leaf tissue with 10 N H2SO4 provides a satisfactory estimate of the endogenous inorganic phosphorus content.

Iodoacetate depression in Xenopus sciatic single nerve fibers

1965 ◽  
Vol 208 (4) ◽  
pp. 720-723 ◽  
Author(s):  
Gordon M. Schoepfle ◽  
Eliska Atkins ◽  
Larry A. Schafer
Keyword(s):  
Membrane Potential ◽  
Maximum Rate ◽  
Nerve Fibers ◽  
Rate Of Change ◽  
Resting Membrane Potential ◽  
Delayed Effect ◽  
Fluid Exchange ◽  
Sodium Conductance ◽  
Single Nerve ◽  
A Chain

Under conditions of continuous fluid exchange at a pH 7.55, a 10-min exposure of Xenopus sciatic single nerve fibers to iodoacetate results in eventual decline in the maximum rate of change of membrane potential, even after a delay of an hour or more during which no changes are apparent. This delayed effect is obtained over an iodoacetate concentration range of 0.1–20.0 mm sodium iodoacetate. Neither the resting membrane potential nor the maximal limiting response obtained during hyperpolarization are affected at a time when iodoacetate has appreciably depressed the spike in the nonpolarized fiber. These findings are taken to indicate that iodoacetate blocks a chain of reactions at a link remote from the process directly concerned with maintenance of the resting level of the sodium conductance. Neither lactate nor pyruvate can be relied on to bring about recovery from the iodoacetate depression.

Increased intracellular Na+ and depolarization favor angiotension tachyphylaxis in rabbit aorta

1991 ◽  
Vol 260 (2) ◽  
pp. H373-H378 ◽  
Author(s):  
E. Frediani-Neto ◽  
E. G. Silva ◽  
T. B. Paiva ◽  
A. C. Paiva
Keyword(s):  
Smooth Muscle ◽  
Membrane Potential ◽  
Angiotensin Ii ◽  
Rabbit Aorta ◽  
Ang Ii ◽  
Free Medium ◽  
Intracellular Na Activity ◽  
Activity Measurements ◽  
Site Of Action ◽  
Rate Limiting

Tachyphylaxis to both angiotensin II (ANG II) and Sar1-ANG II is observed in normal rabbit aorta rings, but helical strips show tachyphylaxis only to Sar1-ANG II, whereas everted rings are not tachyphylactic to either analogue. In normal rings, a good correlation was observed between intraluminal pH and degree of tachyphylaxis to both analogues, suggesting that rate-limiting access of the agonists to their site of action may enhance tachyphylaxis in this preparation. Membrane potential and intracellular Na+ activity measurements, as well as the relaxation by K+ of norepinephrine-contracted preparations in K(+)-free medium, indicated that helical strips are more depolarized than everted rings due to Na+ leakage into the smooth muscle cells. These results suggest that the differences in the degree of tachyphylaxis induced by angiotensin in different rabbit aorta preparations are due to a less accessible site of action in normal rings and to the higher intracellular Na+ and more depolarized state of helical strips relative to everted rings.

scholarly journals Ion Channel Activities in Neural Stem Cells of the Neuroepithelium

2012 ◽  
Vol 2012 ◽  
pp. 1-6 ◽  
Author(s):  
Masayuki Yamashita
Keyword(s):  
Stem Cells ◽  
Membrane Potential ◽  
Neural Stem Cells ◽  
Nuclear Envelope ◽  
Electrical Coupling ◽  
Receptor Activation ◽  
S Phase ◽  
Basal Region ◽  
Neuroepithelial Cell ◽  
Neuroepithelial Cells

During the embryonic development of the central nervous system, neuroepithelial cells act as neural stem cells. They undergo interkinetic nuclear movements along their apico-basal axis during the cell cycle. The neuroepithelial cell shows robust increases in the nucleoplasmic [Ca2+] in response to G protein-coupled receptor activation in S-phase, during which the nucleus is located in the basal region of the neuroepithelial cell. This response is caused by Ca2+release from intracellular Ca2+stores, which are comprised of the endoplasmic reticulum and the nuclear envelope. The Ca2+release leads to the activation of Ca2+entry from the extracellular space, which is called capacitative, or store-operated Ca2+entry. These movements of Ca2+are essential for DNA synthesis during S-phase. Spontaneous Ca2+oscillations also occur synchronously across the cells. This synchronization is mediated by voltage fluctuations in the membrane potential of the nuclear envelope due to Ca2+release and the counter movement of K+ions; the voltage fluctuation induces alternating current (AC), which is transmitted via capacitative electrical coupling to the neighboring cells. The membrane potential across the plasma membrane is stabilized through gap junction coupling by lowering the input resistance. Thus, stored Ca2+ions are a key player in the maintenance of the cellular activity of neuroepithelial cells.

scholarly journals Origin of the membrane potential in plasmodial droplets of Physarum polycephalum. Evidence for an electrogenic pump.

1981 ◽  
Vol 78 (6) ◽  
pp. 637-655 ◽  
Author(s):  
H Kuroda ◽  
R Kuroda
Keyword(s):  
Membrane Potential ◽  
Potassium Cyanide ◽  
Equilibrium Potential ◽  
Ion Pump ◽  
Electrophysiological Studies ◽  
Concentration Changes ◽  
Electrogenic Ion Pump ◽  
Ph Range ◽  
Physarum Plasmodia ◽  
Spherical Droplets

Spherical droplets, derived from Physarum plasmodia by incubation in 10 mM caffeine, seemed to be an excellent system for electrophysiological studies because they were large (less than or equal to 300 micrometer in diameter) and because they tolerated intracellular electrodes filled with 3 M KCl and 10 mM EDTA for a few hours. Intact plasmodia, by contrast, gave valid records for only a few minutes. Under standard conditions ([K+]o = 1 mM, [Na+]o = 5 mM, [Ca++]0 = 0.5 mM, [Mg++]o = 2 mM, and [Cl-]o = 6 mM at pH 7.0), the potential difference across droplet membranes was -80 to -120mV, interior negative. The membrane potential was only slightly sensitive to concentration changes for the above-mentioned ions, and was far negative to the equilibrium diffusion potentials calculated from the known internal contents of K, Na, Ca, Mg, and CL (29.4, 1.6, 3.7, 6.5, and 27.8 mmol/kg, respectively). Variations of external pH did have a strong influence on the membrane potential, yielding a slope of 59 mV/pH between pH 6.5 and 5.5. In this pH range, however, the equilibrium potential for H+ (assuming 6.2 less than or equal to pHi less than or equal to 7.0) was greater than 75 mV positive to the observed membrane potential. Membrane potential was directly responsive to metabolic events, being lowered by potassium cyanide, and by cooling from 25 to 12 degrees C. This ensemble of results strongly indicates that the major component of membrane potential in plasmodial droplets of Physarum is generated by an electrogenic ion pump, probably one extruding H+ ions.

scholarly journals Effect of presynaptic membrane potential on electrical vs. chemical synaptic transmission

2011 ◽  
Vol 106 (2) ◽  
pp. 680-689 ◽  
Author(s):  
Colin G. Evans ◽  
Bjoern Ch. Ludwar ◽  
Timothy Kang ◽  
Elizabeth C. Cropper
Keyword(s):  
Membrane Potential ◽  
Synaptic Transmission ◽  
Electrical Coupling ◽  
Mammalian Brain ◽  
Resting Membrane Potential ◽  
Presynaptic Membrane ◽  
Electrical Transmission ◽  
A Cell ◽  
Peripheral Activation ◽  
Chemical Transmission

The growing realization that electrical coupling is present in the mammalian brain has sparked renewed interest in determining its functional significance and contrasting it with chemical transmission. One question of interest is whether the two types of transmission can be selectively regulated, e.g., if a cell makes both types of connections can electrical transmission occur in the absence of chemical transmission? We explore this issue in an experimentally advantageous preparation. B21, the neuron we study, is an Aplysia sensory neuron involved in feeding that makes electrical and chemical connections with other identified cells. Previously we demonstrated that chemical synaptic transmission is membrane potential dependent. It occurs when B21 is centrally depolarized prior to and during peripheral activation, but does not occur if B21 is peripherally activated at its resting membrane potential. In this article we study effects of membrane potential on electrical transmission. We demonstrate that maximal potentiation occurs in different voltage ranges for the two types of transmission, with potentiation of electrical transmission occurring at more hyperpolarized potentials (i.e., requiring less central depolarization). Furthermore, we describe a physiologically relevant type of stimulus that induces both spiking and an envelope of depolarization in the somatic region of B21. This depolarization does not induce functional chemical synaptic transmission but is comparable to the depolarization needed to maximally potentiate electrical transmission. In this study we therefore characterize a situation in which electrical and chemical transmission can be selectively controlled by membrane potential.

scholarly journals EFFICACY OF PACLOBUTRAZOL UPTAKE THROUGH PECAN LEAVES, YOUNG SHOOTS, AND BARK

HortScience ◽  
1990 ◽  
Vol 25 (8) ◽  
pp. 862c-862
Author(s):  
Gregory L. Reighard ◽  
Harvey M. Jessup
Keyword(s):  
Biological Activity ◽  
Growth Regulator ◽  
Shoot Growth ◽  
Leaf Surface ◽  
Leaf Tissue ◽  
Distilled Water ◽  
Green Wood ◽  
Shoot Tissue ◽  
Abaxial Leaf Surface ◽  
Leaf Flush

Paclobutrazol, a triazole growth regulator, effectively regulates pecan vegetative growth when applied as a soil or trunk drench. However, its absorption and subsequent biological activity in leaves and shoot tissue is not well understood. Terminal shoots from scaffolds of 8-yr-old `Chickasaw' pecan trees were treated with paclobutrazol after leaf flush in mid-May of 1988. Treatments included painting a mixture of 10 mg a.i. paclobutrazol and 1 ml distilled water onto either 1-yr-old wood, green wood, or the abaxial leaf surface. Shoot growth measurements and nut counts were taken in October of 1988 and 1989 on the treated shoots and all shoots arising from them. Paclobutrazol significantly increased the number of nuts per shoot in 1988, but did not affect shoot growth. More nuts were found on shoots from the 1-yr-old wood and leaf treatments than from the control and green wood treatments. In 1989, shoot growth was significantly less in the 2 former than the 2 latter treatments. These data indicate that paclobutrazol was absorbed through the bark of 1-yr-old wood and abaxial leaf tissue and sub-sequently translocated to areas of shoot growth.

Microelectrode measurements of K+ and pH in rabbit gastric glands: effect of histamine

1984 ◽  
Vol 246 (4) ◽  
pp. G433-G444
Author(s):  
K. Kafoglis ◽  
S. J. Hersey ◽  
J. F. White
Keyword(s):  
Membrane Potential ◽  
Ion Exchange ◽  
Basolateral Membrane ◽  
Free Medium ◽  
Na Transport ◽  
K Uptake ◽  
Gastric Glands ◽  
Microelectrode Measurements ◽  
Intracellular K Activity ◽  
K Activity

Conventional and liquid ion-exchange microelectrodes sensitive to K+ or pH were used to examine the response of isolated rabbit gastric glands to histamine. The epithelial cells were impaled across the basolateral membrane. The membrane potential averaged -6.1 +/- 0.6 mV and was unchanged after replacement of medium K+, Cl-, or Na+. The intracellular K+ activity (alpha iK) averaged 41.3 +/- 3.0 mM, indicating K+ accumulation by a factor of 6.8. Active accumulation of K+ was eliminated by ouabain. In contrast, histamine increased K+ activity to 55.3 +/- 3.9 mM. This stimulation was blocked by ouabain. In glands bathed in a Na+-free medium containing ouabain, addition of histamine elevated alpha iK from 12.5 +/- 0.7 to 17.1 +/- 1.1 mM. Isobutylmethylxanthine (10(-4) M) also elevated alpha iK. When impaled with pH-sensitive microelectrodes, glands exposed to histamine exhibited regions of acidity as low as pH 3. Acidification was also produced by histamine after medium Na+ had been replaced with choline. Picoprazole (H 149/94) blocked the effects of histamine on alpha iK and gland pH. The results are consistent with the view that histamine-induced acid secretion by gastric glands is associated with K+ uptake by a mechanism that is independent of Na+ transport but is inhibited by intracellular Na+. This is most likely the H+-K+-ATPase on the secretory surface of the gland cells. Evidence that some tissue K+ is bound or compartmentalized is also discussed.

scholarly journals High-Yield Production of Receptor Binding Domain of SARS-CoV-2 Linked to Bacterial Flagellin in Plants Using Self-Replicating Viral Vector pEff

Plants ◽  
2021 ◽  
Vol 10 (12) ◽  
pp. 2682
Author(s):  
Eugenia S. Mardanova ◽  
Roman Y. Kotlyarov ◽  
Nikolai V. Ravin
Keyword(s):  
Recombinant Protein ◽  
Fusion Protein ◽  
Receptor Binding ◽  
Viral Vector ◽  
Leaf Tissue ◽  
Binding Domain ◽  
Soluble Form ◽  
Leaf Biomass ◽  
Receptor Binding Domain ◽  
Fresh Leaf

The development of recombinant vaccines against SARS-CoV-2 is required to eliminate the COVID-19 pandemic. We reported the expression of a recombinant protein Flg-RBD comprising receptor binding domain of SARS-CoV-2 spike glycoprotein (RBD) fused to flagellin of Salmonella typhimurium (Flg), known as mucosal adjuvant, in Nicotiana benthamiana plants. The fusion protein, targeted to the cytosol, was transiently expressed using the self-replicating vector pEff based on potato virus X genome. The recombinant protein Flg-RBD was expressed at the level of about 110–140 μg per gram of fresh leaf tissue and was found to be insoluble. The fusion protein was purified using metal affinity chromatography under denaturing conditions. To increase the yield of Flg-RBD, the flow-through fraction obtained after loading of the protein sample on the Ni-NTA resin was re-loaded on the sorbent. The yield of Flg-RBD after purification reached about 100 μg per gram of fresh leaf tissue and the purified protein remained soluble after dialysis. The control flagellin was expressed in a soluble form and its yield after purification was about 300 μg per gram of fresh leaf biomass. Plant-produced Flg-RBD protein could be further used for the development of intranasal recombinant mucosal vaccines against COVID-19.

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