Inward-rectifier potassium channels in basolateral membranes of frog skin epithelium

Inward-rectifier K channel: using macroscopic voltage clamp and single- channel patch clamp techniques we have identified the K+ channel responsible for potassium recycling across basolateral membranes (BLM) of principal cells in intact epithelia isolated from frog skin. The spontaneously active K+ channel is an inward rectifier (Kir) and is the major component of macroscopic conductance of intact cells. The current- voltage relationship of BLM in intact cells of isolated epithelia, mounted in miniature Ussing chambers (bathed on apical and basolateral sides in normal amphibian Ringer solution), showed pronounced inward rectification which was K(+)-dependent and inhibited by Ba2+, H+, and quinidine. A 15-pS Kir channel was the only type of K(+)-selective channel found in BLM in cell-attached membrane patches bathed in physiological solutions. Although the channel behaves as an inward rectifier, it conducts outward current (K+ exit from the cell) with a very high open probability (Po = 0.74-1.0) at membrane potentials less negative than the Nernst potential for K+. The Kir channel was transformed to a pure inward rectifier (no outward current) in cell- attached membranes when the patch pipette contained 120 mM KCl Ringer solution (normal NaCl Ringer in bath). Inward rectification is caused by Mg2+ block of outward current and the single-channel current-voltage relation was linear when Mg2+ was removed from the cytosolic side. Whole-cell current-voltage relations of isolated principal cells were also inwardly rectified. Power density spectra of ensemble current noise could be fit by a single Lorentzian function, which displayed a K dependence indicative of spontaneously fluctuating Kir channels. Conclusions: under physiological ionic gradients, a 15-pS inward- rectifier K+ channel generates the resting BLM conductance in principal cells and recycles potassium in parallel with the Na+/K+ ATPase pump.     

氮添加对毛竹林土壤酸性磷酸单酯酶动力学参数的影响

土壤磷酸酶在有机磷矿化和磷循环过程中发挥着重要作用,然而,土壤磷酸酶响应氮(N)沉降的动力学机制仍不清楚。本研究在亚热带毛竹林中设置对照(0)、20(低氮)、40(中氮)和80 g N·hm^-2·a^-1(高氮)4种不同氮添加处理,在氮添加满3年、5年和7年时采集0～15 cm土层土壤样本,测定了土壤化学性质、微生物生物量,并分析了酸性磷酸单酯酶(ACP)的最大反应速率(V_m)、半饱和常数(K_m)和催化效率(K_a)。结果表明: 氮添加显著降低了土壤可溶性有机碳、有效磷和有机磷含量,显著增加了土壤铵态氮、硝态氮含量和V_m,且V_m与有效磷、有机磷和可溶性有机碳含量存在显著相关关系;总体上,氮添加显著提高了K_a;除了在氮添加满5年时高氮处理下K_m显著高于对照外,氮添加对K_m无显著影响,且K_m与有效磷和有机磷含量有显著负相关关系。中、高氮处理对ACP动力学参数的影响大于低氮处理。方差分解分析表明,土壤化学性质的变化而非微生物学性质的变化主导了V_m(47%)和K_m(33%)的变化。总之,氮添加显著影响了毛竹林土壤的基质有效性,通过调控ACP动力学参数(尤其是V_m)进而影响了土壤磷循环。本研究有助于了解氮素富集下土壤微生物调节土壤磷循环的潜在机制,并为全球变化下土壤磷循环模型优化提供重要参数。     

Activation of inwardly rectifying potassium (Kir) channels by phosphatidylinosital-4,5-bisphosphate (PIP2): interaction with other regulatory ligands

All members of the inwardly rectifying potassium channels (Kir1-7) are regulated by the membrane phospholipid, phosphatidylinosital-4,5-bisphosphate (PIP₂). Some are also modulated by other regulatory factors or ligands such as ATP and G-proteins, which give them their common names, such as the ATP sensitive potassium (K_ATP) channel and the G-protein gated potassium channel. Other more non-specific regulators include polyamines, kinases, pH and Na⁺ ions. Recent studies have demonstrated that PIP₂ acts cooperatively with other regulatory factors to modulate Kir channels. Here we review how PIP₂ and co-factors modulate channel activities in each subfamily of the Kir channels.     

Characterization of cocaine-sensitive dopamine uptake in PC12 cells

Dopamine uptake in rat pheochromocytoma (PC12) cells is a carrier-mediated process which follows Michaelis Menten kinetics. Uptake was saturable with an apparent K_m of 0.71 μM for dopamine and a V_max of 3.2 pmol/2 × 10⁵ cells/min. The rank order of potency for various amines was norepinephrine copamine > epinephrine. Uptake increased with increasing temperature and showed a sharp break in the Arrhenius plot at 27.5 C. The Q₁₀ was 1.39 above and 2.95 below 27.5 C. Cocaine inhibited uptake in a dose-dependent manner with a K₁ of 0.97 μM. The presence of cocaine lowered the apparent K_m but did not affect the V_max, indicating competitive inhibition. Tunicamycin inhibited [³H]dopamine accumulation in a dose- and time-dependent fashion suggesting the dopamine uptake site in PC12 cells is an asparagine-linked glycoprotein. Kinetic analysis showed a decrease in V_max but not in the apparent K_m after tunicamycin treatment, consistent with the notion that tunicamycin treatment results in the loss of a substantial amount of active carrier molecules.     

Osmolality influences bistability of membrane potential under hypokalemic conditions in mouse skeletal muscle: an experimental and theoretical study

The membrane potential in mouse skeletal muscle depends on both extracellular osmolality and potassium concentration. These dependencies have been related to two membrane transporters, Na+/K+/2Cl- co-transporter and the inward potassium rectifier channel. To investigate the relation of the Na+/K+/2Cl- co-transporter and the inward potassium rectifier channel in a qualitative way, a combined electrophysiological and modelling approach was used. The experimental results show that the bistability of the membrane potential, which is related to the conductive state of the inward potassium rectifier channel, is shifted to higher extracellular potassium values when medium osmolality is increased. These results are confirmed by the computer simulation calculations for increased co-transporter flux. The combined results indicate that the co-transporter is capable of modulating the conductive state of the inward potassium rectifier channel.     

Micellar catalysis of polyphenol oxidase in AOT/cyclohexane

The catalytic behaviour of mushroom polyphenol oxidase has been studied in dioctylsulphosuccinate (AOT)/cyclohexane reverse micelles. The steady-state conditions were accomplished up to 20 min and 17 μg protein in the assay towards 4-methylcatechol and no loss of specific activity was observed relative to aqueous medium. The pH activity profile of the enzyme was kept in reverse micelles as in water, showing a plateau between 5 and 6.5. The stability of polyphenol oxidase to pH was also studied and about 20% inactivation was found in reverse micelles relative to aqueous medium at neutral pHs. Moreover there was a decrease of stability at acidic pHs. The optimum W_o obtained was 20 and the enzyme was nearly independent of the surfactant concentration at constant W_o.

Kinetic studies of polyphenol oxidase towards several substrates showed that the substrate inhibition by p-cresol and 4-methylcatechol observed in buffer was not kept in AOT/cyclohexane reverse micelles. Moreover, the K_m increased and the catalytic efficiency (V/K_m) of the enzyme decreased as the hydrophobicity of substrates was increased.     

Modelling action potentials and membrane currents of mammalian skeletal muscle fibres in coherence with potassium concentration changes in the T-tubular system

During prolonged activity the action potentials of skeletal muscle fibres change their shape. A model study was made as to whether potassium accumulation and removal in the tubular space is important with respect to those variations. Classical Hodgkin-Huxley type sodium and (potassium) delayed rectifier currents were used to determine the sarcolemmal and tubular action potentials. The resting membrane potential was described with a chloride conductance, a potassium conductance (inward rather than outward rectifier) and a sodium conductance (minor influence) in both sarcolemmal and tubular membranes. The two potassium conductances, the Na-K pump and the potassium diffusion between tubular compartments and to the external medium contributed to the settlement of the potassium concentration in the tubular space. This space was divided into 20 coupled concentric compartments. In the longitudinal direction the fibre was a cable series of 56 short segments. All the results are concerned with one of the middle segments. During action potentials, potassium accumulates in the tubular space by outward current through both the delayed and inward rectifier potassium conductances. In between the action potentials the potassium concentration decreases in all compartments owing to potassium removal processes. In the outer tubular compartment the diffusion-driven potassium export to the bathing solution is the main process. In the inner tubular compartment, potassium removal is mainly effected by re-uptake into the sarcoplasm by means of the inward rectifier and the Na-K pump. This inward transport of potassium strongly reduces the positive shift of the tubular resting membrane potential and the consequent decrease of the action potential amplitude caused by inactivation of the sodium channels. Therefore, both potassium removal processes maintain excitability of the tubular membrane in the centre of the fibre, promote excitation-contraction coupling and contribute to the prevention of fatigue. Received: 5 May 1998 / Revised version: 27 October 1998 / Accepted: 19 January 1999     

A negative resistance region underlies the triggering property of membrane potential in human T-lymphocytes

Steady-state current-voltage relationships (SSCVRs) of the plasma membrane of human T-lymphocytes were studied at the physiological temperature of 37°C by using the whole-cell patch-clamp technique. SSCVRs displayed a characteristic N-like shape with a negative resistance region (NRR) in a voltage range of −45 to −35 mV. The majority of cells assayed revealed SSCVR patterns crossing the V-axis at three points (in mV): V₁ = −55 to −45, V₂ = −40 to −35, V₃ = −30 to −10. SSCVRs of T-cells activated by phytohaemagglutinin (48–96 h) also displayed NRR, but crossed the V-axis at one point only (V₁ = −55 to −60 mV). It implies the possibility of two stable levels of membrane potential (V₁ and V₃) for the resting T-cells, but only one (V₁) for activated T-cells. These data thus account for the triggering property of T-cell membrane potential previously reported. The NRR can be explained on the basis of the Hodgkin-Huxley type n⁴j model of K⁺ channel kinetics. According to the model the possibility for a membrane to have on or two stable levels of membrane potential depends on the ratio of selective K⁺ conductance to non-selective leaky conductance (G_k/G_leak). The steady-state level of K⁺ conductance in resting T-lymphocytes proved to be sensitive to Ca²⁺. Buffering Ca²⁺ ions from either external or internal solution resulted in an appreciable increase in K⁺ conductance. The possibility for membrane potential have two stable levels of membrane potential in connection with the Ca²⁺ dependence of K⁺ conductance was supposed to be important for Ca²⁺-signalling during T-cell activation.     

Circadian Variation in the Uptake of Tryptophan by Cortical Synaptosomes of the Rat Brain

The kinetics of the high affinity uptake system for L-tryptophan (L-Try)have been measured over 24 hr in cortical synaptosome preparations of rat brain. Both the K_m and V_max, of the uptake process showed a statistically significant 24 hr variation. The highest K_m value, 6.71 ± 10^-5 M, was measured at the beginning of the light phase and the lowest value, 4.23 ± 10^-5 M, 6 hr into the dark phase. V_max was highest at the end of the dark phase (10.43 nmol/mg/5 min) and lowest (4.80 nmol/mg/5 min) 3 hr into the dark phase. In contrast, there was no variation over 24 hr in the V_max/K_m ratio. These results suggest that the high affinity uptake process serves to ensure a constant rate of L-tryptophan entry into the neuron in the face of circadian or ultradian variations in extracellular concentration of tryptophan.     

Kinetic characterization of sulphur-containing excitatory amino acid uptake in primary cultures of neurons and astrocytes

The uptake of the neuroactive sulphur amino acids -cysteine sulphinate, -cysteate, -homocysteine sulphinate and -homocysteate was investigated in astrocytes cultured from the prefrontal cortex; in neurons, cultured from cerebral cortex; and, in granule cells, cultured from cerebellum. It was shown that each amino acid acted as a substrate for a plasma membrane transporter in both neurons and astrocytes. Astrocytes and neurons exhibited a high-affinity uptake for -cysteine sulphinate and -cysteate with K_m values ranging from 14–100 μM, and a low-affinity uptake for -homocysteine sulphinate and -homocysteate, with K_m values ranging from 225–1210 μM. The uptake of all transmitter candidates studied was partially sodium-dependent. This sodium-dependency was most evident at low (< 100 μM) concentrations of each substrate. The apparent uptake measured in the absence of sodium was included as a component in corrections made for non-saturable influx. With the exception of -cysteine sulphinate, uptake of each sulphur amino acid was greatest in astrocytes, with V_max values ranging between 15–32 nmol min⁻¹ mg⁻¹ cell protein. Moreover, the uptake of each sulphur amino acid in cerebellar granule cells (V_max values ranging between 10–25 nmol min⁻¹ mg⁻¹ cell protein) was consistently greater than that in cerebral cortex neurons (V_max values ranging between 1.5–6 nmol min⁻¹ mg⁻¹ cell protein).     

葛根素对大鼠心室肌细胞动作电位及钾通道电流的影响

目的:观察葛根素对大鼠心室肌细胞动作电位及钾通道电流的影响。方法:用常规微电极方法记录大鼠心室肌细胞动作电位,用全细胞膜片钳技术记录游离心室肌细胞钾离子流。结果:不同浓度的葛根素均能延长大鼠心室肌细胞动作电位时程(APD)及抑制内向整流钾电流,具有明显的浓度依赖关系。结论:葛根素延长APD,抑制内向整流钾电流,可能是其抗心律失常的机制。     

P, P-bis(5′-adenosyl)triphosphate (Ap3A) as a substrate and a product of mammalian tryptophanyl-tRNA synthetase

Bovine tryptophanyl-tRNA synthetase (TrpRS, E.C. 6.1.1.2) is unable to catalyze in vitro formation of Ap₄A in contrast to some other aminoacyl-tRNA synthetases. However, in the presence of -tryptophan, ATP-Mg²⁺ and ADP the enzyme catalyzes the Ap₃A synthesis via adenylate intermediate. Ap₃A (not Ap₄A) may serve as a substrate for TrpRS in the reaction of E·(Trp AMP) formation and in the tRNA^Trp charging. The K_m value for Ap₃A was higher than the K_m for ATP (approx. 1.00 vs. 0.22 mM) and V_max was 3 times lower than for ATP. The Zn²⁺-deficient enzyme catalyzes Ap₃A synthesis in the absence of exogenous ADP due to ATPase activity of Zn²⁺-deprived TrpRS. The inability of mammalian TrpRS to synthesize Ap₄A, might be considered as a molecular tool preventing the removal of Zn²⁺ due to chelation by Ap₄A and therefore preserving the enzyme activity.     

G protein control of potassium channel activity in a mast cell line

Using the patch-clamp technique, we studied regulation of potassium channels by G protein activators in the histamine-secreting rat basophilic leukemia (RBL-2H3) cell line. These cells normally express inward rectifier K+ channels, with a macroscopic whole-cell conductance in normal Ringer ranging from 1 to 16 nS/cell. This conductance is stabilized by including ATP or GTP in the pipette solution. Intracellular dialysis with any of three different activators of G proteins (GTP gamma S, GppNHp, or AlF-4) completely inhibited the inward rectifier K+ conductance with a half-time for decline averaging approximately 300 s after "break-in" to achieve whole-cell recording. In addition, with a half-time averaging approximately 200 s, G protein activators induced the appearance of a novel time-independent outwardly rectifying K+ conductance, which reached a maximum of 1-14 nS. The induced K+ channels are distinct from inward rectifier channels, having a smaller single-channel conductance of approximately 8 pS in symmetrical 160 mM K+, and being more sensitive to block by quinidine, but less sensitive to block by Ba2+. The induced K+ channels were also highly permeable to Rb+ but not to Na+ or Cs+. The current was not activated by the second messengers Ca2+, inositol 1,4,5-trisphosphate, inositol 1,3,4,5-tetrakisphosphate, or by cyclic AMP-dependent phosphorylation. Pretreatment of cells with pertussis toxin (0.1 microgram/ml for 12-13 h) prevented this current's induction both by guanine nucleotides and aluminum fluoride, but had no effect on the decrease in inward rectifier conductance. Since GTP gamma S is known to stimulate secretion from patch-clamped rat peritoneal mast cells, it is conceivable that K+ channels become inserted into the plasma membrane from secretory granules. However, total membrane capacitance remained nearly constant during appearance of the K+ channels, suggesting that secretion induced by GTP gamma S was minimal. Furthermore, pertussis toxin had no effect on secretion triggered by antigen, and triggering of secretion before electrical recording failed to induce the outward K+ current. Finally, GTP gamma S activated the K+ channel in excised inside-out patches of membrane. We conclude that two different GTP-binding proteins differentially regulate two subsets of K+ channels, causing the inward rectifier to close and a novel K+ channel to open when activated.     

Bioconversion of phospholipids by immobilized phospholipase A2

Phospholipase A₂ selectively hydrolyses the ester linkage at the sn-2 position of phospholipids forming lysocompounds. This bioconversion has importance in biotechnology since lysophospholipids are strong bioemulsifiers. The aim of the present work was to study the kinetic behaviour and properties of immobilized phospholipase A₂ from bee venom adsorbed into an ion exchange support. The enzyme had high affinity for CM-Sephadex^® support and the non-covalent interaction was optimum at pH 8. The activity of immobilized phospholipase A₂ was comparatively evaluated with the soluble enzyme using a phospholipid/Triton X-100 mixed micelle as assay system. The immobilized enzyme showed high retention activity and excellent stability under storage. The activity of the immobilized system remained almost constant after several cycles of hydrolysis. Immobilized phospholipase A₂ was less sensitive to pH changes compared to soluble form. The kinetic parameters obtained (V_max 883.4 μmol mg⁻¹ min⁻¹ and a K_m 12.9 mM for soluble form and V_max = 306 μmol mg⁻¹ min⁻¹ and a K_m = 3.9 for immobilized phospholipase A₂) were in agreement with the immobilization effect. The results obtained with CM-Sephadex^®-phospholipase A₂ system give a good framework for the development of a continuous phospholipid bioconversion process.     

Internal Na+ and Mg2+ blockade of DRK1 (Kv2.1) potassium channels expressed in Xenopus oocytes. Inward rectification of a delayed rectifier

Delayed rectifier potassium channels were expressed in the membrane of Xenopus oocytes by injection of rat brain DRK1 (Kv2.1) cRNA, and currents were measured in cell-attached and inside-out patch configurations. In intact cells the current-voltage relationship displayed inward going rectification at potentials > +100 mV. Rectification was abolished by excision of membrane patches into solutions containing no Mg2+ or Na+ ions, but was restored by introducing Mg2+ or Na+ ions into the bath solution. At +50 mV, half- maximum blocking concentrations for Mg2+ and Na+ were 4.8 +/- 2.5 mM (n = 6) and 26 +/- 4 mM (n = 3) respectively. Increasing extracellular potassium concentration reduced the degree of rectification of intact cells. It is concluded that inward going rectification resulting from voltage-dependent block by internal cations can be observed with normally outwardly rectifying DRK1 channels.     

A reversible NO complex of Fe(TIM): an S = 1/2FeNO nitrosyl

[Fe(TIM)(CH₃CN)₂](PF₆)₂ (1) (TIM = 2,3,9,10-tetramethyl-1,4,8,11-tetraazacyclodeca-1,3,8,10-tetraene) forms a complex with NO reversibly in CH₃CN (53±1% converted to the NO complex) or 60% CH₃OH/40% CH₃CN (81±1% conversion). Quantitative NO complexation occurs in H₂O or CH₃OH solvents. The EPR spectrum of [Fe(TIM)(solvent)NO]²⁺ in frozen 60/40 CH₃OH/CH₃CN at 77 K shows a three line feature at g=2.01, 1.99 and 1.97 of an S=1/2FeNO⁷ ground state. The middle line exhibits a three-line N-shf coupling of 24 G indicating a six-coordinate complex with either CH₃OH or CH₃CN as a ligand trans to NO. In H₂O [Fe(TIM)(H₂O)₂]²⁺ undergoes a slow decomposition, liberating 2,3-butanedione, as detected by ¹H NMR in D₂O, unless a π-acceptor axial ligand, L=CO, CH₃CN or NO is present. An equilibrium of 1 in water containing CH₃CN forms [Fe(TIM)(CH₃CN)(H₂O)]²⁺ which has a formation constant K_CH3CN=320 M⁻¹. In water K_NOK_CH3CN since NO completely displaces CH₃CN. [Fe(TIM)(CH₃CN)₂]²⁺ binds either CO or NO in CH₃CN with K_NO/K_CO=0.46, sigificantly lower than the ratio for [Fe^II(hemes)] of 1100 in various media. A steric influence due to bumping of β-CH₂ protons of the TIM macrocycle with a bent S=1/2 nitrosyl as opposed to much lessened steric factors for the linear Fe---CO unit is proposed to explain the lower K_NO/K_CO ratio for the [Fe(TIM)(CH₃CN)]²⁺ adducts of NO or CO. Estimates for formation constants with [Fe(TIM)]²⁺ in CH₃CN of K_NO=80.1 M⁻¹ and K_CO=173 M⁻ are much lower than to hemoglobin (where K_NO=2.5×10¹⁰ M⁻¹ and K_CO=2.3×10⁷) due to a reversal of steric factors and stronger π-backdonation from [Fe^II(heme)] than from [Fe^II(TIM)(CH₃CN)]²⁺.     

Characterization of Dir: a putative potassium inward rectifying channel in Drosophila

Potassium channels vary in their function and regulation, yet they maintain a number of important features - they are involved in the control of potassium flow, cell volume, cell membrane resting potential, cell excitability and hormone release. The potassium (K(+)) inward rectifier (Kir) superfamily of channels are potassium selective channels, that are sensitive to the concentration of K(+) ions. They are termed inward rectifiers since they allow a much greater K(+) influx than efflux. There are at least seven subfamilies of Kir channels, grouped according to sequence and functional similarities (Curr. Opin. Neurobiol. 5 (1995) 268; Annu. Rev. Physiol. 59 (1997) 171). While numerous Kir channels have been discovered in a variety of organisms, Drosophila inward rectifier (Dir) is the first putative inward rectifier to be studied in Drosophila. In fact, there are only three genes (including Dir) encoding putative inward rectifiers in the Drosophila genome. Though there are other known potassium channels in Drosophila such as ether-a-go-go and shaker, most are voltage-gated channels. As an important first step in characterizing Kir channels in Drosophila, we initiated studies on Dir.     

芝麻与花生间作对芝麻功能叶光合荧光特性的影响

为了明确芝麻与花生间作提高芝麻产量的光合机理,于2017—2018年设芝麻与花生3∶6间作(IC 3∶6)、2∶4间作(IC 2∶4)、芝麻单作(SS)、花生单作(SP) 4个处理,研究了间作对芝麻功能叶气体交换参数、光合-光强和光合-CO₂响应曲线、快速叶绿素荧光诱导动力学曲线的影响.结果表明:间作芝麻产量的偏土地当量比大于1/3;与单作芝麻相比,芝麻与花生间作提高了芝麻功能叶的光饱和点(I_sat)、光饱和时的净光合速率(P_{n max})、最大电子传递速率(J_max)、磷酸丙糖利用率(TPU)、Rubisco最大羧化速率(V_{c max});提高了单位面积吸收(ABS/CS_o)、捕获(TR_o/CS_o)和电子传递(ET_o/CS_o)的能量、反应中心的数目(RC/CS_m)和传递到PSⅠ末端的量子产额(RE_o/CS_o);降低了可变荧光F_k占F_j-F_o振幅的比例(W_k)和可变荧光F_j占F_p-F_o振幅的比例(V_j),提高了PSⅡ反应中心捕获光能转化为电能的效率(Ψ_o)、PSⅡ将电子传递到PSⅠ受体侧末端的效率(Ψ_Ro)、电子传递链的电子传递效率(δ_R)、PSⅠ光化学活性(ΔI/I_o)和光系统间协调性(Φ_PSⅠ/PSⅡ).IC 3∶6下芝麻功能叶的净光合速率(P_n)、气孔导度(g_s)、蒸腾速率(T_r)、P_{n max}、J_max、V_{c max}、TPU、Ψ_o、Ψ_Ro和δ_R均高于IC 2∶4,其中P_n、g_s和T_r差异显著.这说明间作芝麻具有明显产量间作优势关键在于间作能促进其功能叶对光能的吸收、传递与转化,提高电子传递链性能,增强PSⅠ、PSⅡ性能和两者间协调性及CO₂羧化固定能力,从而提高净光合速率,其中IC 3∶6优于IC 2∶4.     

Paradoxical effect of ethanol on potassium channel currents and cell survival in cerebellar granule neurons

Transient exposure to ethanol (EtOH) results in a massive neurodegeneration in the developing brain leading to behavioral and cognitive deficits observed in fetal alcohol syndrome. There is now compelling evidence that K⁺ channels play an important role in the control of programmed cell death. The aim of the present work was to investigate the involvement of K⁺ channels in the EtOH-induced cerebellar granule cell death and/or survival. At low and high concentrations, EtOH evoked membrane depolarization and hyperpolarization, respectively. Bath perfusion of EtOH (10 mM) depressed the I _A (transient K⁺ current) potassium current whereas EtOH (400 mM) provoked a marked potentiation of the specific I _K (delayed rectifier K⁺ current) current. Pipette dialysis with GTPγS or GDPβS did not modify the effects of EtOH (400 mM) on both membrane potential and I _K current. In contrast, the reversible depolarization and slowly recovering inhibition of I _A induced by EtOH (10 mM) became irreversible in the presence of GTPγS. EtOH (400 mM) induced prodeath responses whereas EtOH (10 mM) and K⁺ channel blockers promoted cell survival. Altogether, these results indicate that in cerebellar granule cells, EtOH mediates a dual effect on K⁺ currents partly involved in the control of granule cell death.     

Quick and effective hyperpolarization of the membrane potential in intact smooth muscle cells of blood vessels by synchronization modulation electric field

Blood vessel dilation starts from activation of the Na/K pumps and inward rectifier K channels in the vessel smooth muscle cells, which hyperpolarizes the cell membrane potential and closes the Ca channels. As a result, the intracellular Ca concentration reduces, and the smooth muscle cells relax and the blood vessel dilates. Activation of the Na/K pumps and the membrane potential hyperpolarization plays a critical role in blood vessel functions. Previously, we developed a new technique, synchronization modulation, to control the pump functions by electrically entraining the pump molecules. We have applied the synchronization modulation electric field noninvasively to various intact cells and demonstrated the field-induced membrane potential hyperpolarization. We further applied the electric field to blood vessels and investigated the field induced functional changes of the vessels. In this paper, we report the results in a study of the membrane potential change in the smooth muscle cells of mesenteric blood vessels in response to the oscillating electric field. We found that the synchronization modulation electric field can effectively hyperpolarize the muscle membrane potential quickly in seconds under physiological conditions.