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1.
N. S. Kavok M. Y. Malyukina I. A. Borovoy E. N. Obukchova S. A. Klimov 《Cell biochemistry and biophysics》2013,67(2):763-771
Early events in individual hepatocytes of rat activated by adrenaline (10?6M) and phenylephrine (10?5M) have been investigated by quantitative image microfluorometry and microspectrofluorometry. Cationic DiOC2 and anionic SqSC4 probes have been used for image analysis and transmembrane potential (ΔΨ p) estimation in real-time studies. Fluorescence spectra resulting from the accumulation of dyes in single cells were recorded. Based on the mean fluorescence intensity, the magnitude of ΔΨ p was calculated by Nernst equation adapted for lipophilic cationic probes. DiOC2 has revealed that both hormones induce biphasic hyperpolarization of hepatocytes membrane with α-agonist phenylephrine causing ΔΨ p changes at higher amplitude. The first increase of ΔΨ p within 2 and 5 min (ΔΔΨ p = ?8.6 ± 4.2 mV) apparently related to Na+/K+-ATPase activation by the Ca2+-mobilizing hormone. The second peak of hyperpolarization (ΔΔΨ p = ?13.2 ± 3.2 mV) between 25 and 30 min, after a transient decrease of ΔΨ p (ΔΔΨ p = 10.9 ± 4.3 mV) over 15 min experiment, probably is mediated by phenylephrine stimulating action on K+-channels. K+ channel blocker (Ba2+ or 4-aminopyridine) as well as elevating of extracellular K+ prevented the hyperpolarization. Modulation of PLD-dependent signal transduction pathway by 0.4 % butanol had a weak influence on the first increase of ΔΨ p but it abolished the second phase of hyperpolarization. That points to PLD involvement in the ΔΨ p fluctuations mediated by K+-channels in response to phenylephrine. Based on SqSC4, fluorescent parameters estimation of relative changes of ΔΨ p revealed similar character of time dependence with two phases of hyperpolarization. Synchronic fluctuation of ΔΨ p determined by oppositely charged probes demonstrate that the quantitative microfluorometry allows to evaluate slight ΔΨ p changes separately from ΔΨ m in non-excitable individual cells at the short-term hormone action. 相似文献
2.
Leon Pape 《生物化学与生物物理学报:生物膜》1982,686(2):225-232
Changes in fluorescence intensity of thiodicarbocyanine, DiS-C3(5), were correlated with direct microelectrode potential measurements in red blood cells from Amphiuma means and applied qualitatively to evaluate the effects of extracellular Ca2+, K+ and pH on the membrane potential of human red cells. Increasing extracellular [Ca2+] from 1.8 to 15 mM causes a K+-dependent hyperpolarization and decrease in fluorescence intensity in Amphiuma red cells. Both the hyperpolarization and fluorescence change disappear when the temperature is raised from 17 to 37°C. No change in fluorescence intensity is observed in human red cells with comparable increase in extracellular Ca2+ in the temperature range 5–37°C. Increasing the extracellular pH, however, causes human red cells to respond to an increase in extracellular Ca2+ with a significant but temporary loss in fluorescence intensity. This effect is blocked by EGTA, quinine or by increasing extracellular [K+], indicating that at elevated extracellular pH, human erythrocytes respond to an increase in extracellular Ca2+ with an opening of K+ channels and associated hyperpolarization of the plasma membrane. 相似文献
3.
Effects of Light on the Membrane Potential of Protoplasts from Samanea saman Pulvini : Involvement of K Channels and the H -ATPase 总被引:1,自引:0,他引:1
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Rhythmic light-sensitive movements of the leaflets of Samanea saman depend upon ion fluxes across the plasma membrane of extensor and flexor cells in opposing regions of the leaf-movement organ (pulvinus). We have isolated protoplasts from the extensor and flexor regions of S. saman pulvini and have examined the effects of brief 30-second exposures to white, blue, or red light on the relative membrane potential using the fluorescent dye, 3,3′-dipropylthiadicarbocyanine iodide. White and blue light induced transient membrane hyperpolarization of both extensor and flexor protoplasts; red light had no effect. Following white or blue light-induced hyperpolarization, the addition of 200 millimolar K+ resulted in a rapid depolarization of extensor, but not of flexor protoplasts. In contrast, addition of K+ following red light or in darkness resulted in a rapid depolarization of flexor, but not of extensor protoplasts. In both flexor and extensor protoplasts, depolarization was completely inhibited by tetraethylammonium, implicating channel-mediated movement of K+ ions. These results suggest that K+ channels are closed in extensor plasma membranes and open in flexor plasma membranes in darkness and that white and blue light, but not red light, close the channels in flexor plasma membranes and open them in extensor plasma membranes. Vanadate treatment inhibited hyperpolarization in response to blue or white light, but did not affect K+ -induced depolarization. This suggests that white or blue light-induced hyperpolarization results from activation of the H+ -ATPase, but this hyperpolarization is not the sole factor controlling the opening of K+ channels. 相似文献
4.
Jose Luis De La Vega-Beltran Claudia Sánchez-Cárdenas Darío Krapf Enrique O. Hernandez-González Eva Wertheimer Claudia L. Trevi?o Pablo E. Visconti Alberto Darszon 《The Journal of biological chemistry》2012,287(53):44384-44393
Mammalian sperm are unable to fertilize the egg immediately after ejaculation; they acquire this capacity during migration in the female reproductive tract. This maturational process is called capacitation and in mouse sperm it involves a plasma membrane reorganization, extensive changes in the state of protein phosphorylation, increases in intracellular pH (pHi) and Ca2+ ([Ca2+]i), and the appearance of hyperactivated motility. In addition, mouse sperm capacitation is associated with the hyperpolarization of the cell membrane potential. However, the functional role of this process is not known. In this work, to dissect the role of this membrane potential change, hyperpolarization was induced in noncapacitated sperm using either the ENaC inhibitor amiloride, the CFTR agonist genistein or the K+ ionophore valinomycin. In this experimental setting, other capacitation-associated processes such as activation of a cAMP-dependent pathway and the consequent increase in protein tyrosine phosphorylation were not observed. However, hyperpolarization was sufficient to prepare sperm for the acrosome reaction induced either by depolarization with high K+ or by addition of solubilized zona pellucida (sZP). Moreover, K+ and sZP were also able to increase [Ca2+]i in non-capacitated sperm treated with these hyperpolarizing agents but not in untreated cells. On the other hand, in conditions that support capacitation-associated processes blocking hyperpolarization by adding valinomycin and increasing K+ concentrations inhibited the agonist-induced acrosome reaction as well as the increase in [Ca2+]i. Altogether, these results suggest that sperm hyperpolarization by itself is key to enabling mice sperm to undergo the acrosome reaction. 相似文献
5.
Yuan Gao Ying Yang Qigang Guan Xuefeng Pang Haishan Zhang Dingyin Zeng 《Molecular and cellular biochemistry》2010,338(1-2):59-68
The large conductance Ca2+-activated K+ (BK) channel, abundantly expressed in vascular smooth muscle cells, plays a critical role in controlling vascular tone. Activation of BK channels leads to membrane hyperpolarization and promotes vasorelaxation. BK channels are activated either by elevation of the intracellular Ca2+ concentration or by membrane depolarization. It is also regulated by a diversity of vasodilators and vasoconstrictors. Interleukin-1β (IL-1β) is one of the cytokines that play important roles in the development and progression of a variety of cardiovascular diseases. The effects of IL-1β on vascular reactivity are controversial, and little is known about the modulation of BK channel function by IL-1β. In this study, we investigated how IL-1β modulates BK channel function in cultured arterial smooth muscle cells (ASMCs), and examined the role of H2O2 in the process. We demonstrated that IL-1β had biphasic effects on BK channel function and membrane potential of ASMCs, that is both concentration and time dependent. IL-1β increased BK channel-dependent K+ current and hyperpolarized ASMCs when applied for 30 min. While long-term (24–48 h) treatment of IL-1β resulted in decreased expression of α-subunit of BK channel, suppressed BK channel activity, decreased BK channel-dependent K+ current and depolarization of the cells. H2O2 scavenger catalase completely abolished the early effect of IL-1β, while it only partly diminished the long-term effect of IL-1β. These results may provide important molecular mechanisms for therapeutic strategies targeting BK channel in inflammation-related diseases. 相似文献
6.
Illumination of an Aplysia giant neuron evokes a membrane hyperpolarization which is associated with a membrane conductance increase of 15%. The light response is best elicited at 490 nM: the neuron also has an absorption peak at this wavelength. At the resting potential (-50 to -60 mV) illumination evokes an outward current in a voltage-clamped cell. This current reverses sign very close to EK calculated from direct measurements of internal and external K+ activity. Increases in external K+ concentration shift the reversal potential of the light-evoked response by the same amount as the change in EK. Decreases in external Na+ or Cl- do not affect the response. Therefore, the response is attributed to an increase in K+ conductance. Pressure injection of Ca2+ into this neuron also hyperpolarizes the cell membrane. This effect is also due largely to an increase in K+ conductance. The light response after Ca2+ injection does not appear to be altered. Pressure injection of EGTA abolished or greatly reduced the light response. The effect was reversible. We suggest that light acts upon a single pigment in this neuron, releasing Ca2+ which in turn increases K+ conductance, thereby hyperpolarizing the neuronal membrane. 相似文献
7.
Mice lacking the gene for suppressor of cytokine signaling 1 (SOCS1) show defective homeostasis of T lymphocytes due to accumulation of CD8+ T cells, resulting at least partly from dysregulated IL-15 signaling. IL-15 alone does not stimulate proliferation of naïve CD8 T cells, but can synergize with IL-21 to induce proliferation, suggesting a potential role for IL-21 in the defective homeostasis of CD8+ T lymphocytes in SOCS1−/− mice. Since IL-21 strongly induced SOCS1 mRNA in CD8+ T cells, we investigated whether SOCS1 regulates their response to IL-21. CD8+ T cells isolated from SOCS1-deficient mice proliferated vigorously in response to IL-21 + IL-15. In CD8+ T lymphocytes expressing transgenic TCR, IL-21 + IL-7 provided a stronger stimulus to naïve cells whereas IL-15 + IL-21 potently stimulated memory cells. Compared to truly naïve or memory cells, SOCS1−/− H-Y TCR+ CD8+ T cells displayed CD44loLy6ChiCD122intCD127lo partial memory phenotype and exhibited stronger response to IL-15 + IL-21 than truly naïve cells. In SOCS1−/− CD8+ T cells, IL-21 caused greater reduction in IL-15 threshold for activation in a dose-dependent manner. SOCS1 deficiency did not modulate IL-21Rα expression or sensitivity to IL-21, but delayed the loss of IL-21-induced phospho-STAT3 signal. These results show that SOCS1 is a critical regulator of IL-21 signaling in CD8+ T cells, and support the notion that sustained IL-21 signaling might also contribute to the aberrant T cell homeostasis in SOCS1-deficient mice. 相似文献
8.
The effect of adrenaline on the K+-activated hyperpolarization of frog skeletal muscle fibres was studied. The amplitude of K+-activated hyperpolarization, which was produced when the external K+ concentration was changed from 0 to 2 mM, was markedly increased in the presence of adrenaline. In the presence of ouabain (1 × 10?5 M), which completely and reversibly eliminated the K+-activated hyperpolarization, adrenaline caused no significant changes in both the membrane potential and conductance under the condition where the K+-activated hyperpolarization was supposed to be produced. These results suggested that adrenaline accelerated the electrogenic Na+ pump which produced the K+-activated hyperpolarization. 相似文献
9.
On the Theory of Ion Transport across the Nerve Membrane: V. Two Models for the Cole-Moore K+ Hyperpolarization Delay 总被引:2,自引:1,他引:1
Two illustrative molecular models, designed to explain the Cole-Moore K+ hyperpolarization delay, are proposed and analyzed. Both introduce a process supplementary to the usual Hodgkin-Huxley (HH) one for a K+ channel. In both cases the new process becomes involved as a consequence of the conditioning hyperpolarization of the membrane and would account for the observed delay time in the K+ current after depolarization to near ENa. The first model uses adsorption or desorption of phospholipid molecules on the surface of the assumed protein K+ channel or gate. The second model involves the translocation of the charged subunits of the channel in the hyperpolarizing electric field. 相似文献
10.
Ying Cui Lihui Zheng Ming Jiang Ru Jia Xiao Zhang Qishan Quan Guiqin Du Dongjin Shen Xiaodan Zhao Wenying Sun Hongwei Xu Lijuan Huang 《Molecular biology reports》2013,40(11):6437-6442
Microparticles (MPs) are vesicles released from activated or apoptotic cells. MP derive from various cells, most notably platelets, but also leucocytes, lymphocytes, erythrocytes, and endothelial cells. The aim of this study was to investigate endothelial MP (EMP), platelet MP (PMP), lymphocyte MP and monocyte MP and TF-positive MPs (TF+ MPs) in patients with coronary heart disease (CHD), and to evaluate the correlation of these MPs with Interleukin-6 (IL-6) and C-reactive protein (CRP). Different cell-derived MPs and TF+ MPs were analyzed by flow cytometry in 40 patients with myocardial infarction (MI), 30 unstable angina (UA), 20 stable angina (SA) and 20 healthy individuals, and IL-6 and CRP were determined by ELISA and special protein analyzer, respectively. Compared with SA and control, EMP and PMP was significantly elevated in MI and UA (P < 0.001), and TF+ MPs was significantly elevated in MI and UA (P < 0.001). EMP and PMP correlated with IL-6 (r = 0.822, P < 0.001 and r = 0.567, P < 0.001; respectively) or CRP level (r = 0.597, P < 0.001 and r = 0.66, P < 0.001; respectively). Different cell-derived MPs in CHD may indicate the different pathophysiological changes in vessels, and MPs may both participate in the development of thrombosis and enhance the vascular inflammation. 相似文献
11.
G.A. Volkov 《BBA》1973,314(1):83-92
Transient hyperpolarization of the external cytoplasmatic membrane may be observed on rapid illumination of the Nitella flexilis cell. Several important properties of that response make the latter similar to a considerable degree to the excitation response.The condition for transient hyperpolarization is the normal functioning of the electron transport chain conjugated with non-cyclic photophosphorylation.The value of the membrane potential at the moment of hyperpolarization of the external cytoplasmic membrane, is determined by the difference in the electrochemical potential of HCO3? or H+. This state of the plasmalemma supplements the two other known states: normal and depolarized (excited), when the main ions determining membrane potential are K+ and Cl?. 相似文献
12.
Summary The fluorescent anionic dye, bisoxonol, and flow cytometry have been used to monitor changes in the membrane potential of rat thymocytes exposed to the B subunit of cholera toxin. The B subunit induced a rapid hyperpolarization, which was due to activation of a Ca2+-sensitive K+ channel. Reduction of extracellular Ca2+ to <1 m by the addition of [ethylenebis(oxyethylenenitrilo)]tetraacetic acid immediately abolished the hyperpolarization caused by the B subunit. Cells treated with quinine and tetraethylammonium lost their ability to respond to the B subunit, whereas 4-aminopyridine did not have any effect. Thus, calcium-sensitive and not voltage-gated K+ channels appeared to be responsible for the hyperpolarization. The results of ion substitution experiments indicated that extracellular Na+ was not essential for changes in membrane potential. Further studies with ouabain, amiloride and furosemide demonstrated that electrogenic Na+/K+ ATPase, Na+/H+ antiporter and Na+/K+/Cl– cotransporter, respectively, were not involved in the hyperpolarization process induced by the B subunit. Thus, crosslinking of several molecules of ganglioside GM1 on the cell surface of rat thymocytes by the pentavalent B subunit of cholera toxin modulated plasma membrane permeability to K+ by triggering the opening of Ca2+-sensitive K+ channels. A role for gangliosides in regulating ion permeability would have important implications for the function of gangliosides in various cellular phenomena. 相似文献
13.
Ho S. Oh Beom K. Choi Young H. Kim Don G. Lee Sunhee Hwang Myoung J. Lee Sang H. Park Yong-Soo Bae Byoung S. Kwon 《PloS one》2015,10(5)
4-1BB (CD137), a member of the tumor necrosis factor receptor superfamily (TNFRSF), is primarily expressed on activated T cells and is known to enhance proliferation of T cells, prevent activation-induced cell death, and promote memory formation of CD8+ T cells. In particular, it is well acknowledged that 4-1BB triggering preferentially enhances the expansion of CD8+ T cells rather than CD4+ T cells, but the underlying mechanism remains unclear. Here we found that 4-1BB triggering markedly increased IL-2Rα (CD25) and IL-2 expressions of CD8+ T cells but minimally for CD4+ T cells. Proliferation of CD8+ T cells was moderately enhanced by direct 4-1BB triggering in the absence of signaling through IL-2Rα/IL-2 interactions, but further promoted in the presence of IL-2Rα/IL-2 interactions. Among the TNFRSF members including OX40, GITR, CD30, and CD27, 4-1BB was superior in the ability to induce IL-2Rα expression on CD8+ T cells. When the primary and secondary expansions of CD8+ T cells in vivo were examined by adoptively transferring OVA-specific CD8+ T cells along with the treatment with agonistic anti-4-1BB and/or antagonistic anti-CD25 F(ab’)2 mAb, 4-1BB triggering enhanced both primary and secondary expansion of CD8+ T cells in vivo, and the 4-1BB effects were moderately suppressed in primary expansion while completely abolished in secondary expansion of OVA-specific CD8+ T cells by blocking IL-2Rα. These results suggest that 4-1BB-mediated increases of IL-2Rα and IL-2 prolong the effects of transient TCR- and 4-1BB-mediated signaling in CD8+ T cells, and that 4-1BB triggering preferentially enhances the expansion of CD8+ T cells through the amplification of autocrine IL-2/IL-2R signaling loop. 相似文献
14.
Carmen Beltrán Esmeralda Rodríguez-Miranda Gisela Granados-González Lucia García de De la Torre Takuya Nishigaki Alberto Darszon 《Developmental biology》2014
Zinc (Zn2+) has been recently recognized as a crucial element for male gamete function in many species although its detailed mechanism of action is poorly understood. In sea urchin spermatozoa, Zn2+ was reported as an essential trace ion for efficient sperm motility initiation and the acrosome reaction by modulating intracellular pH (pHi). In this study we found that submicromolar concentrations of free Zn2+ change membrane potential (Em) and increase the concentration of intracellular Ca2+ ([Ca2+]i) and cAMP in Lytechinus pictus sperm. Our results indicate that the Zn2+ response in sperm of this species mainly involves an Em hyperpolarization caused by K+ channel activation. The pharmacological profile of the Zn2+-induced hyperpolarization indicates that the cGMP-gated K+ selective channel (tetraKCNG/CNGK), which is crucial for speract signaling, is likely a main target for Zn2+. Considering that Zn2+ also induces [Ca2+]i fluctuations, our observations suggest that Zn2+ activates the signaling cascade of speract, except for an increase in cGMP, and facilitates sperm motility initiation upon spawning. These findings provide new insights about the role of Zn2+ in male gamete function. 相似文献
15.
Melanie J. McCoy Anna K. Nowak Robbert G. van der Most Ian M. Dick Richard A. Lake 《Cancer immunology, immunotherapy : CII》2013,62(3):529-539
There is a complex interplay between the immune system and a developing tumor that is manifest in the way that the balance of T cell subsets in the local tumor environment reflects clinical outcome. Tumor infiltration by CD8+ T cells and regulatory T cells (Treg) is associated with improved and reduced survival, respectively, in many cancer types. However, little is known of the prognostic value of immunological parameters measured in peripheral blood. In this study, peripheral CD8+ T cells and Treg from 43 patients with malignant mesothelioma or advanced non-small-cell lung cancer scheduled to commence palliative chemotherapy were assessed by flow cytometry and evaluated for association with patient survival. Patients had a higher proportion of peripheral Treg, proliferating CD8+ T cells and CD8+ T cells with an activated effector phenotype compared with age-matched healthy controls. Higher proportions of Treg and proliferating CD8+ T cells were both associated with poor survival in univariate analyses (hazard ratio [HR] 3.81, 95 % CI 1.69–8.57; p < 0.01 and HR 2.86, 95 % CI 1.26–6.50; p < 0.05, respectively). CD8+ T cell proliferation was independently predictive of reduced survival in multivariate analysis (HR 2.58, 95 % CI 1.01–6.61; p < 0.05). These findings suggest that peripheral CD8+ T cell proliferation can be a useful prognostic marker in patients with thoracic malignancies planned for palliative chemotherapy. 相似文献
16.
Julio C. Chávez José L. de la Vega-Beltrán Jessica Escoffier Pablo E. Visconti Claudia L. Trevi?o Alberto Darszon Lawrence Salkoff Celia M. Santi 《PloS one》2013,8(4)
Unlike most cells of the body which function in an ionic environment controlled within narrow limits, spermatozoa must function in a less controlled external environment. In order to better understand how sperm control their membrane potential in different ionic conditions, we measured mouse sperm membrane potentials under a variety of conditions and at different external K+ concentrations, both before and after capacitation. Experiments were undertaken using both wild-type, and mutant mouse sperm from the knock-out strain of the sperm-specific, pH-sensitive, SLO3 K+ channel. Membrane voltage data were fit to the Goldman-Hodgkin-Katz equation. Our study revealed a significant membrane permeability to both K+ and Cl− before capacitation, as well as Na+. The permeability to both K+ and Cl− has the effect of preventing large changes in membrane potential when the extracellular concentration of either ion is changed. Such a mechanism may protect against undesired shifts in membrane potential in changing ionic environments. We found that a significant portion of resting membrane potassium permeability in wild-type sperm was contributed by SLO3 K+ channels. We also found that further activation of SLO3 channels was the essential mechanism producing membrane hyperpolarization under two separate conditions, 1) elevation of external pH prior to capacitation and 2) capacitating conditions. Both conditions produced a significant membrane hyperpolarization in wild-type which was absent in SLO3 mutant sperm. Hyperpolarization in both conditions may result from activation of SLO3 channels by raising intracellular pH; however, demonstrating that SLO3-dependent hyperpolarization is achieved by an alkaline environment alone shows that SLO3 channel activation might occur independently of other events associated with capacitation. For example sperm may undergo stages of membrane hyperpolarization when reaching alkaline regions of the female genital tract. Significantly, other events associated with sperm capacitation, occur in SLO3 mutant sperm and thus proceed independently of hyperpolarization. 相似文献
17.
Boron induces hyperpolarization of sunflower root cell membranes and increases membrane permeability to k 总被引:5,自引:1,他引:4
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Although many studies have alluded to a role for boron (B) in membrane function, there is little evidence for a direct effect of B on the plasmalemma of higher plant cells. These studies were conducted to demonstrate, by electrophysiological techniques, a direct effect of B on the membrane potential (Em) of sunflower (Helianthus annuus [L.], cv Mammoth Grey Stripe) root tip cells and to determine if the response to B occurs rapidly enough to account for the previously observed effects of B on ion uptake. By inserting a glass microelectrode into an individual cell in the root tip, the Em of the cell was determined in basal salt medium (BSM), pH 6.0. The perfusion solution surrounding the root tissue was then changed to BSM + 50 micromolar H3BO3, pH 6.0. The exposure to B induced a significant plasmalemma hyperpolarization in sunflower root cells within 20 minutes. After just 3 minutes of exposure to B, the change in Em was already significantly different from the negligible change in Em observed over time in root cells never exposed to B. Membrane hyperpolarization could be caused by a stimulation of the proton pump or by a change in the conductance of one or more permeable ions. Since B has been shown to affect K+ uptake by plants, the electrophysiological techniques described above were used to determine if B has an effect on membrane permeability to K+, and could thereby lead to an increased diffusion potential. When sunflower root tips were pretreated in 50 micromolar B for 2 hours, cell membranes exhibited a significantly greater depolarization with each 10-fold increase in external [K+] than minus-B cells. Subsequent studies demonstrated that the depolarization due to increased external [K+] was also significantly greater when tissue was exposed to B at the same time as the 10-fold increase in [K+], indicating that the effect of B on K+ permeability was immediate. Analysis of sunflower root tips demonstrated that treatment in 50 micromolar B caused a significantly greater accumulation of K+ after 48 hours. The B-induced increase in K+ uptake may cause a subsequent stimulation of the H+-ATPase (proton pump) and lead to the observed hyperpolarization of root cell membranes. Alternatively, B may stimulate the proton pump, with the subsequent hyperpolarization resulting in an increased driving force for K+ influx. 相似文献
18.
The aim of this work was to examine the effects of changes in external K+ concentration (K
o
) around its physiological value, of various K+ channels blockers, including internal Cs+, of vacuolar H+-ATPase inhibitors and of the protonophore CCCP on the resting potential and the voltage-dependent K+ current of differentiated neuroblastoma x glioma hybrid NG108-15 cells using the whole-cell patch-clamp technique. The results are as follows: (i) under standard conditions (K
o
=5 mm) the membrane potential was –60±1 mV. It was unchanged when K
o
was decreased to 1 mm and was depolarized by 4±1 mV when Ko was increased to 10 mm. (ii) Internal Cs+ depolarized the membrane by 21±3 mV. (iii) The internal application of the vacuolar H+-ATPase inhibitors N-ethylmaleimide (NEM), NO
3
–
and bafilomycin A1 (BFA) depolarized the membrane by 15±2, 18±2 and 16±2 mV, respectively, (iv) When NEM or BFA were added to the internal medium containing Cs+, the membrane was depolarized by 45±1 and 42±2 mV, respectively. (v) The external application of CCCP induced a transient depolarization followed by a prolonged hyperpolarization. This hyperpolarization was absent in BFA-treated cells. The voltage-dependent K+ current was increased at negative voltages and decreased at positive voltages by NEM, BFA and CCCP. Taken together, these results suggest that under physiological conditions, the resting potential of NG108-15 neuroblastoma cells is maintained at negative values by both voltage-dependent K+ channels and an electrogenic vacuolar type H+-ATPase.This work was supported by a grant from INSERM (CRE 91 0906). 相似文献
19.
Tom L. Van Belle Hans Dooms Tom Boonefaes Xiao-Qing Wei Georges Leclercq Johan Grooten 《PloS one》2012,7(9)
Due to its critical role in NK cell differentiation and CD8+ T cell homeostasis, the importance of IL-15 is more firmly established for cytolytic effectors of the immune system than for CD4+ T cells. The increased levels of IL-15 found in several CD4+ T cell-driven (auto-) immune diseases prompted us to examine how IL-15 influences murine CD4+ T cell responses to low dose TCR-stimulation in vitro. We show that IL-15 exerts growth factor activity on both CD4+ and CD8+ T cells in a TCR-dependent and Cyclosporin A-sensitive manner. In CD4+ T cells, IL-15 augmented initial IL-2-dependent expansion and once IL-15Rα was upregulated, IL-15 sustained the TCR-induced expression of IL-2/15Rβ, supporting proliferation independently of secreted IL-2. Moreover, IL-15 counteracts CD4+ T cell suppression by a gradually expanding CD25HighCD4+ T cell subset that expresses Foxp3 and originates from CD4+CD25+ Tregs. These in vitro data suggest that IL-15 may dramatically strengthen the T cell response to suboptimal TCR-triggering by overcoming an activation threshold set by Treg that might create a risk for autoimmune pathology. 相似文献
20.
45Ca2+ fluxes across the plasma membrane of zygotes of the fucoid alga, Pelvetia fastagiata (J. Ag.) De Toni, were studied in artificial sea waters of various potassium concentrations. Except for two cases, hyperpolarization of the cell membrane (with low [K+]) increases, and depolarization (with high [K+]) decreases the influx of Ca2+ over the range of [K+] studied (1–100 mM). The fractional increases of influx during hyperpolarization are close to the fractional increases in membrane potential but the decreases during depolarization are much smaller than those in membrane potential. In two anomalous cases, the influxes of 45Ca2+ at a potassium concentration of 30 mM were about 20% higher than the control value instead of being 10% lower.The effluxes of 45Ca2+ are increased by both hyperpolarization and by depolarization. On balance (and excepting the two anomalous cases) the net result of hyperpolarization should be to increase and that of depolarization to decrease intracellular [Ca2+]. 相似文献