Enhancement of barium- and cesium-induced adrenal catecholamine release by lidocaine

Summary Catecholamine release evoked from isolated perfused bovine adrenals by Ba⁺² or Cs⁺ is enhanced by lidocaine or by a calcium-free medium. The action of Cs⁺ therefore differs from that of K⁺ or Rb⁺ in adrenal medulla. Divalent and monovalent metallic cations of relatively large atomic weight like Ba⁺² and Cs⁺, probably penetrate the cell more easily than small highly charged ions and act intracellularly to cause adrenal catecholamine release. Local anesthetics and calcium-free media may allow greater influx of Ba⁺² and Cs⁺ into adrenomedullary cells.Acknowledgment. This work was supported by NIH, grant No. AM16153.     

Effects of barium on separately recorded fast and slow PIII responses in bullfrog retina

Investigation of Ba2+ effects on fast and slow PIII responses in isolated bullfrog retina revealed that Ba2+ suppressed slow PIII completely with little effect on fast PIII. A light-induced [K+]0 decrease in the photoreceptor layer was observed in spite of Ba2+ perfusion, indicating the suppressive action of Ba2+ on the K+ conductance of the Müller cell membrane.     

Blockade of voltage-dependent and Ca2+-dependent K+ current components by internal Ba2+ in molluscan pacemaker neurons

Summary Internal Ba²⁺ blocks both the Ca-dependent component, as well as the voltage-dependent component of the K⁺ current in molluscan pacemaker neurons.     

The effects of triaminopyrimidine on the short circuit current and transmembrane potentials of the isolated rat visceral yolk sac in vitro

Summary Intracellular potentials in the cells from 17.5-day old rat visceral yolk sacs were measured by a glass microelectrode. When penetrated from the maternal side, the cells have potentials of about 50.2±1.9 mV (inside negative) which were reduced by increasing the external K⁺ concentration and increased by removing Na⁺ ions from the bathing fluid. Triaminopyrimidine (TAP) which inhibited Na⁺ transport caused a dose-dependent depolarization of the cell membrane. The depolarization was dependent on the presence of extracellular Ca²⁺ ions. It is proposed that TAP may inhibit Na⁺ transport by increasing the intracellular concentration of calcium ions.This work was supported by the University of Hong Kong (grant number 335. 034.5105).Acknowledgment. Triaminopyrimidine was synthesized by Dr. Barbara Roth of the Wellcome Research Laboratories.     

Ionic mechanisms in pancreatic β cell signaling

The function and survival of pancreatic β cells critically rely on complex electrical signaling systems composed of a series of ionic events, namely fluxes of K⁺, Na⁺, Ca²⁺ and Cl^? across the β cell membranes. These electrical signaling systems not only sense events occurring in the extracellular space and intracellular milieu of pancreatic islet cells, but also control different β cell activities, most notably glucose-stimulated insulin secretion. Three major ion fluxes including K⁺ efflux through ATP-sensitive K⁺ (K_ATP) channels, the voltage-gated Ca²⁺ (Ca_V) channel-mediated Ca²⁺ influx and K⁺ efflux through voltage-gated K⁺ (K_V) channels operate in the β cell. These ion fluxes set the resting membrane potential and the shape, rate and pattern of firing of action potentials under different metabolic conditions. The K_ATP channel-mediated K⁺ efflux determines the resting membrane potential and keeps the excitability of the β cell at low levels. Ca²⁺ influx through Ca_V1 channels, a major type of β cell Ca_V channels, causes the upstroke or depolarization phase of the action potential and regulates a wide range of β cell functions including the most elementary β cell function, insulin secretion. K⁺ efflux mediated by K_V2.1 delayed rectifier K⁺ channels, a predominant form of β cell K_V channels, brings about the downstroke or repolarization phase of the action potential, which acts as a brake for insulin secretion owing to shutting down the Ca_V channel-mediated Ca²⁺ entry. These three ion channel-mediated ion fluxes are the most important ionic events in β cell signaling. This review concisely discusses various ionic mechanisms in β cell signaling and highlights K_ATP channel-, Ca_V1 channel- and K_V2.1 channel-mediated ion fluxes.     

Selective blockade of components of potassium activation inMyxicola axons

Summary The K⁺ conductance inMyxicola giant axons activates in two phases which are pharmacologically separable. The fast phase of K⁺ activation is specifically inhibited by 4-aminopyridine and by the substitution of D₂O for H₂O. We suggestMyxicola giant axons, like the amphibian node of Ranvier, may possess more than one variety of K⁺ channel.     

Immobilization of chicken liver fructose 1,6-bisphosphatase on CNBr-activated Sepharose

Summary Chicken liver fructose 1,6-bisphosphatase is readily immobilized on CNBr-activated Sepharose. The immobilization alters some enzymatic properties. They include broader pH activity curve, loss of activation by K⁺ or NH ₄ ⁺ , increased resistance to inactivation by trypsin, decreased sensitivity to AMP inhibition, and loss of cooperative interaction among AMP-binding sites. The immobilized enzyme retains about 38% or 19% of the specific activity of the native enzyme when the activity is measured in the absence or presence of K⁺, resepctively.This work was supported by grant RR-8006 from the General Research Branch, Division of Research Resources, NIH (USA).     

Vanadate: Non-selective inhibition of transepithelial transport of Na+, H+ and water

Summary In the isolated urinary bladder of the toad, 10^–5–10^–4M orthovanadate produces inhibition of the active transport of Na⁺ and H⁺ ions as well as of antidiuretic hormone-mediated osmotic flow of water. Since transport of H⁺ ions and osmotic water flow are not inhibited when (Na⁺+K⁺)-ATPase is inhibited by ouabain, biological actions of vanadate are not necessarily related to inhibition of (Na⁺+K⁺)-ATPase.This research was supported by grant AM-14915 from the National Institutes of Helath.     

Differences in membrane ion transport between hepatocytes from the periportal and the pericentral areas of the liver lobule

We studied the Na⁺/K⁺ pump, Na⁺/K⁺ ATPase activity, and oxygen consumption (QO₂) in hepatocytes isolated from the periportal (PH) and pericentral (CH) regions of the liver lobule, to provide an insight into the functional properties of these cells. Na⁺/K⁺ pump activity was determined using⁸⁶Rb⁺ (a functional analog of K⁺) and ouabain, a specific inhibitor of this transport system. Our results indicate the the Na⁺/K⁺, pump and Na⁺/K⁺ ATPase activity are significantly lower in CH than in PH, although basal ouabain-sensitive (OS) QO₂ was negligible in both of these cell preparations. However, OSQO₂ was significantly lower in CH than in PH when the Na⁺/K⁺ pump was activated using the ionophore nystatin in a Na⁺-containing medium. These results indicate that the differences in membrane ion transport exist between hepatocytes from different locations of the liver lobule.     

Blockade by zinc ions of K+-induced contraction and calcium in guinea pigTaenia coli

Preincubation with 0.3 mM Zn²⁺ markedly inhibited both the tonic response and Ca²⁺ binding at low affinity sites induced by K⁺ (60 mM), with smaller effects on the phasic response and the high affinity Ca²⁺ sites, inTaenia coli. However, when the muscle was kept in Zn²⁺-containing medium following the first stimulation with the K⁺, the phasic response and the high affinity Ca²⁺ sites were more severely inhibited during the second stimulation with K⁺. This probably indicates that Zn²⁺ reduced the tonic tension response to K⁺ mainly by inhibiting Ca²⁺ influx at the cell membranes ofTaenia coli. However, when Zn²⁺ is continuously present, Ca²⁺ is not supplied at the storage sites and is not available for the phasic response to a second stimulation with K⁺.     

Effects of inhibition and stimulation of Na+−K+ active transport on the resting membrane input conductance of the guinea-pig ventricle

Summary The effects of inhibition by ouabain and stimulation by high frequency drive of the sarcolemmal Na⁺–K⁺ active transport system on the resting input conductance (g_i) of guinea-pig ventricular muscles were determined. Although both pump inhibition and stimulation were associated with changes in electrophysiological properties of the muscles, neither had a significant effect on g_i.Supported by a grant from the North Carolina Heart Association.     

Dissociation between compensatory renal growth and induction of (Na++K+-ATPase in rat kidney after uninefrectomy

Zusammenfassung Nach Uninephrektomie und bilateraler Adrenalektomie entsteht eine Dissoziation zwischen der Nierenvergrösserung und der (Na⁺+K⁺)-ATPase Aktivität. Es besteht wahrscheinlich kein Zusammenhang zwischen der Nierenvergrösserung und der Induktion von (Na⁺+K⁺)-ATPase. Auch in der vergrösserten Niere sind die Nebennierenhormone notwendig, um die (Na⁺+K⁺)-ATPase zu erhalten.

---

---

The work was supported by a grant from P. Carl Petersen's Fund. Mrs.Birthe Bagge Hansen provided excellent technical assistance.     

The effects of manganese and barium on the cardiac pacemaker current,i f,in rabbit sino-atrial node myocytes

Summary The isolation of ionic fluxes contributing to electric currents through cell membranes often requires block of other undesired components which can be achieved, among others, by divalent cations. Mn²⁺ and Ba²⁺ are often used, for example, to block Ca and K currents. Here we have investigated the effects of these two cations on the properties of the hyperpolarization-activated pacemaker current i_f, in rabbit sino-atrial node myocytes, as obtained by voltage clamp analysis. We find that 2 mM Mn²⁺ shifts the i_f activation curve by 3.2±0.3 mV towards more positive values. However, when 1 mM Ba²⁺ is also added, the positive shift is more than halved (1.3±0.2 mV). We find, too, that in the absence of blocking cations the ACh-induced i_f inhibition is slightly higher than in their presence. These results indicate that the alteration of i_f kinetic properties by Ba²⁺ plus Mn²⁺-containing solutions is minimal.     

A histochemical demonstration of the Na++K+-ATPase activity in the thyroid and the effect of cyclic adenosine monophosphate (c-AMP)

Summary With a suitable modification of the Farquhar and Palade technique the Na⁺+K⁺-ATPase activity in guineapig thyroid is demonstrated. The addition of c-AMP (5×10^–6 M or 1.5×10^–5 M) to the incubation media produced an apparent intensification of the Na⁺+K⁺-ATPase activity in the thyroid.This work was supported by a grant from ZMNU of Serbia.     

Direct inhibition of phospholipid scrambling activity in erythrocytes by potassium ions

The exposure of phosphatidylserine (PS) at the cell surface plays a critical role in blood coagulation and serves as a macrophage recognition moiety for the engulfment of apoptotic cells. Previous observations have shown that a high extracellular [K⁺] and selective K⁺ channel blockers inhibit PS exposure in platelets and erythrocytes. Here we show that the rate of PS exposure in erythrocytes decreases by ~50% when the intracellular [K⁺] increases from 0 to physiological concentrations. Using resealed erythrocyte membranes, we further show that lipid scrambling is inducible by raising the intracellular [Ca²⁺] and that K⁺ ions have a direct inhibitory effect on this process. Lipid scrambling in resealed ghosts occurs in the absence of cell shrinkage and microvesicle formation, processes that are generally attributed to Ca²⁺-induced lipid scrambling in intact erythrocytes. Thus, opening of Ca²⁺-sensitive K⁺ channels causes loss of intracellular K⁺ that results in reduced intrinsic inhibitory effect of these ions on scramblase activity. Received 11 September 2008; received after revision 17 October 2008; accepted 27 October 2008     

The effect of physostigmine on (Na++K+)-ATPase activity in different rat brain regions

Summary The activity of (Na⁺+K⁺)-ATPase and acetylcholine esterase were folloed in rat brain cerebral cortex, caudate, thalamus, hippocampus and medulla after i.v. administration of physostigmine. Both enzymes were found to be inhibited in a dose-dependent manner. The most pronounced inhibition of (Na⁺+K⁺)-ATPase was found in caudate. where the highest activity of acetylcholine esterase is found.These studies were supported by a grant from the Union of Science of Republic Serbia, No. 40404-14.     

Diffusion loading conditions determine recovery of protein synthesis in electroporated P3X63 Ag8 cells

Summary Using the suspension cell line P3X63 Ag8 we have studied the impact of the composition of the diffusion medium on cellular protein synthesis under standard electroporation conditions in TBS-Na. This buffer contains the high saline concentration usually present in electroporation-mediated DNA transfection. Electroporation in the presence of TBS-Na resulted in an immediate shut-off of protein synthesis, even though both FITC-dextran (M_r 40 kD) and Semliki Forest virus core protein (M_r 33 kD) were incorporated efficiently into the cytoplasm across the electropores at 0°C. Subsequent resealing of the pores was completed after a 5-min incubation at 37°C. When compared with control cells, overall protein synthesis of electroporated cells recovered slowly to resume a 30% activity after 1 h of incubation at 37°C. We have determined optimal conditions for diffusion loading (which necessitates the presence of ATP, GTP, amino acids, K⁺, Mg²⁺, and Ca²⁺) and resealing (in the presence of K⁺, Mg²⁺, and Ca²⁺), leading to a full and lasting recovery of protein synthesis within 5 min after pore closure.     

The (Na++K+)-ATPase activity in brain of quaking mice

Summary The (Na⁺+K⁺)- and Mg²⁺-dependent ATPase distribution in several brain areas has been investigated in Quaking mutant mice characterized by myelin deficiency. A marked decrease of (Na⁺+K⁺)-ATPase activity has been found in limbic structures, hypothalamus and cerebellum. The Mg²⁺-dependent activity did not change. A possible involvement of the impairment of the (Na⁺+K⁺)-ATPase activity in the seizure susceptibility of this mice is discussed.Chargée de Recherche au CNRS.     

Conotoxins of the O-superfamily affecting voltage-gated sodium channels

The venoms of predatory cone snails harbor a rich repertoire of peptide toxins that are valuable research tools, but recently have also proven to be useful drugs. Among the conotoxins with several disulfide bridges, the O-superfamily toxins are characterized by a conserved cysteine knot pattern: C-C-CC-C-C. While ω-conotoxins and κ-conotoxins block Ca²⁺ and K⁺ channels, respectively, the closely related δ- and μO-conotoxins affect voltage-gated Na⁺ channels (Na_v channels). δ-conotoxins mainly remove the fast inactivation of Na_v channels and, thus, functionally resemble long-chain scorpion α-toxins. μO-conotoxins are functionally similar to μ-conotoxins, since they inhibit the ion flow through Na_v channels. Recent results from functional and structural assays have gained insight into the underlying molecular mechanisms. Both types of toxins are voltage-sensor toxins interfering with the voltage-sensor elements of Na_v channels. Received 27 December 2006; received after revision 30 January 2007; accepted 19 February 2007     

Studies on the transmembrane ion currents in the smooth-muscle cells of the gastric fundus

Summary Under voltage-clamp conditions fast Ca²⁺-inward and early K⁺-outward currents were recorded from the smooth-muscle cells of the gastric fundus. It is assumed that the less electrical excitability of these cells is due to the early activation of the outward current.