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.     

Isolation of sodium-complexing polypeptides from mammalian blood and cardiac muscle

Summary Two distinct polypeptides have been isolated from rat heart and ox blood. They are both found to be effective in forming complexes with sodium ions, and it is suggested that they may have a function in stabilizing sodium ion activity.Acknowledgments. We are indebted to ProfessorG. A. Gilbert, F.R.S. and to Dr.C. J. Lote for helpful discussion, and to the Royal Society for a Grant-in-aid which in part supported the investigation.     

The Na/K pump,resting potential and selective permeability in canine Purkinje fibres at physiologic and room temperatures

All mammalian cells maintain a resting potential generated by ions moving down concentration gradients. In excitable cells, the inside potential is negative relative to outside. In order to maintain this electrochemical gradient, the sodium potassium (Na/K) pump actively transports out three sodium ions for every two potassium ions it brings in. This process generates a net outward current and thus hyperpolaizes the resting potential. I employed dihydroouabain (DHO) to inhibit the Na/K pump and thus measure its contribution to the resting potential. It contributed 9.0 mV at 34°C and 3.8 mV at 25°C. The P_K/P_Na ratios were calculated at both temperatures before and after subtracting the Na/K pump contribution. These ratios also suggested a decreased contribution of the Na/K pump under hypothermia. Taken together, these results suggest that the pump contribution to the resting potential is more significant at physiologic temperatures (34°C) than at room temperature (25°C), and that estimates of selective permeability can only be accurately obtained after assessing and eliminating the Na/K pump contribution to the resting potential.     

Décharge catalytique du proton en présence de séléno-cystine et de nickel

Summary It has been found that seleno-cystine, reduced to seleno-cysteine and bound in ligand form to nickel ion, produces catalytic hydrogen discharge in slight acid media. This discharge occurs in the region of a catalytic prewave located at more positive potentials (E_1/2=–1.14 V, S.C.E.) compared with the normal wave of H₃O+(E_1/2=–1,67 V, S.C.E.).     

Interaction of BW755C,a potent inhibitor of lipoxygenase and cyclo-oxygenase,with mitochondrial cytochrome oxidase

Summary The interaction between BW755C (3-amino-1-[m-(trifluoromethyl)phenyl]-2-pyrazoline), a potent inhibitor of both lipoxygenase and cyclo-oxygenase, and respiratory chain in mitochondria and electron transport particles (ETP) from rat livers was examined. BW755C accelerated the oxygen uptake by mitochondria without the addition of substrate for the respiratory chain. Spectrophotometric study revealed that BW755C was quickly oxidized by cytochrome oxidase in mitochondria to a compound possessing an absorption maximum at 524 nm. p-Phenylenediamine (p-diaminobenzene, PPDA), which, like BW755C, serves as an electron donor to cytoschrome oxidase, was shown to inhibit the generation of active oxygen in macrophages; the inhibition was stronger than that of BW755C. These results strongly suggest that the oxidative conversion of BW755C by mitochondrial cytochrome oxidase is associated with its potentially inhibitory action on the active oxygen-generating system in phagocytes.The authors are indebted to Dr M. Hori, Gifu College of Pharmacy and to Dr Y. Orii, Kyoto University for their kind supplies of BW755C and pure cytochrome oxidase, respectively.     

Beziehung zwischen Nebenniere und aktivem Kationentransport

Summary The active sodium transport through the membrane of coldstored red cells of adrenalectomised rats is significantly decreased, whereas the rate of active exchange of potassium is not influenced by adrenalectomy. Cardiac glycosides likewise slow down only the active sodium exchange in the blood of both the normal and adrenalectomised animals in contrast to their action on human red cells.     

Cardiac cellular electrophysiology: past and present

The time-course of the cardiac action potential can be accounted for in terms of ionic currents crossing the cell membranes. Depolarizing current is carried by Na+ or Ca2+ entering the cells, repolarizing current by K+ leaving the cells. Membrane permeability for the passive movement of these ions is thought to be voltage-dependent as well as time-dependent. Net transfer of charge may also result from active transport, 2 Na+ out against 1 K+ in; or coupled exchange, 3 or 4 Na+ in against 1 Ca2+ out. This review follows the path by which present-day knowledge has been reached. It also gives a few examples to illustrate that electrophysiology has provided concepts useful to clinical cardiology.     

Effect of prolactin on human red cell sodium transport

Summary Incubation of red cells with higher concentrations of prolactin in vitro enhanced the cellular sodium level and produced a significant reduction in erythrocyte membrane adenosine triphosphatase activity. This effect was dose and time-dependent. It is the result of an inhibition of the active sodium pump similar to that produced by ouabain, suggesting altered red cell function and electrolyte balance in hyperprolactinemic states.Acknowledgments. Ovine prolactin was generously supplied by Ferring (Malmö, Sweden) and NPA, NIH (Maryland, USA). Thanks are also due to Prof. S. Dutta, Wayne State University, Detroit, for the gift, of ouabain octahydrate and Dr M. Ramachandran and Prof. S. Ramakrishnan, Department of Biochemistry, JIPMER, Pondicherry, for their encouragement and keen interest in this study.     

Regulation of the cardiac sodium pump

In cardiac muscle, the sarcolemmal sodium/potassium ATPase is the principal quantitative means of active transport at the myocyte cell surface, and its activity is essential for maintaining the trans-sarcolemmal sodium gradient that drives ion exchange and transport processes that are critical for cardiac function. The 72-residue phosphoprotein phospholemman regulates the sodium pump in the heart: unphosphorylated phospholemman inhibits the pump, and phospholemman phosphorylation increases pump activity. Phospholemman is subject to a remarkable plethora of post-translational modifications for such a small protein: the combination of three phosphorylation sites, two palmitoylation sites, and one glutathionylation site means that phospholemman integrates multiple signaling events to control the cardiac sodium pump. Since misregulation of cytosolic sodium contributes to contractile and metabolic dysfunction during cardiac failure, a complete understanding of the mechanisms that control the cardiac sodium pump is vital. This review explores our current understanding of these mechanisms.     

The functional role of sodium taurocholate cotransporting polypeptide NTCP in the life cycle of hepatitis B,C and D viruses

Chronic hepatitis B, C and D virus (HBV, HCV and HDV) infections are a major cause of liver disease and cancer worldwide. Despite employing distinct replication strategies, the three viruses are exclusively hepatotropic, and therefore depend on hepatocyte-specific host factors. The sodium taurocholate co-transporting polypeptide (NTCP), a transmembrane protein highly expressed in human hepatocytes that mediates the transport of bile acids, plays a key role in HBV and HDV entry into hepatocytes. Recently, NTCP has been shown to modulate HCV infection of hepatocytes by regulating innate antiviral immune responses in the liver. Here, we review the current knowledge of the functional role and the molecular and cellular biology of NTCP in the life cycle of the three major hepatotropic viruses, highlight the impact of NTCP as an antiviral target and discuss future avenues of research.     

Similarities between sodium channels in excitable membranes and in epithelia.

The inhibitory effects of the pyrazine derivative, amiloride, on sodium transport in an amphibian epithelium has been studied as a function of pH. It is concluded that the charged (guanidinium) group interacts with a negatively charged acid grouping in the membrane. Similarities between sodium channels in excitable membranes and epithelia are highlighted.     

Similarities between sodium channels in excitable membranes and in epithelia

Summary The inhibitory effects of the pyrazine derivative, amiloride, on sodium transport in an amphibian epithelium has been studied as a function of pH. It is concluded that the charged (guanidinium) group interacts with a negatively charged acid grouping in the membrane. Similarities between sodium channels in excitable membranes and epithelia are highlighted.     

Die Isolierung von Ribonukleinsäuren aus menschlichen Knochenmarkzellen

Summary By use of bentonite and sodium dodecylsulfate, which bind or inhibit ribonucleases, pure RNA preparations were obtained from human bone marrow. Phenol extraction at 4 °C yielded cytoplasmic RNA, which could be separated into 3 main peaks by sucrose density gradient centrifugation. Sedimentation coefficients of these peaks were about 30S, 19S, and 5S, respectively. Re-extraction of the phenolic interphase at 65° gave a polydisperse ribonucleic acid, sedimenting between 2S and 19S. This RNA is characterized by template activity and is supposed to be biologically active m-RNA.     

Structural variations in wheat HKT1;5 underpin differences in Na<Superscript>+</Superscript> transport capacity

An important trait associated with the salt tolerance of wheat is the exclusion of sodium ions (Na⁺) from the shoot. We have previously shown that the sodium transporters TmHKT1;5-A and TaHKT1;5-D, from Triticum monoccocum (Tm) and Triticum aestivum (Ta), are encoded by genes underlying the major shoot Na⁺-exclusion loci Nax1 and Kna1, respectively. Here, using heterologous expression, we show that the affinity (K _m) for the Na⁺ transport of TmHKT1;5-A, at 2.66 mM, is higher than that of TaHKT1;5-D at 7.50 mM. Through 3D structural modelling, we identify residues D⁴⁷¹/a gap and D⁴⁷⁴/G⁴⁷³ that contribute to this property. We identify four additional mutations in amino acid residues that inhibit the transport activity of TmHKT1;5-A, which are predicted to be the result of an occlusion of the pore. We propose that the underlying transport properties of TmHKT1;5-A and TaHKT1;5-D contribute to their unique ability to improve Na⁺ exclusion in wheat that leads to an improved salinity tolerance in the field.     

Contribution of the Na+/K+-pump to the membrane potential

The inward movement of sodium ions and the outward movement of potassium ions are passive and the reverse movements against the electrochemical gradients require the activity of a metabolism-driven Na+/K+-pump. The activity of the Na+/K+-pump influences the membrane potential directly and indirectly. Thus, the maintenance of a normal electrical function requires that the Na+/K+-pump maintain normal ionic concentrations within the cell. The activity of the Na+/K+-pump also influences the membrane potential directly by generating an outward sodium current that is larger when the Na+/K+-pump activity is greater. The activity of the Na+/K+-pump is regulated by several factors including the intracellular sodium concentration and the neuromediators norepinephrine and acetylcholine. The inhibition of the Na+/K+-pump can lead indirectly to the development of inward currents that may cause repetitive activity. Therefore, the Na+/K+-pump modifies the membrane potential in different ways both under normal and abnormal conditions and influences in an essential way many cardiac functions, including automaticity, conduction and contraction. Key words. Active transport of ions; cardiac tissues; electroneutral and electrogenic Na+/K/-pump; control of Na+/K+-pump; normal and abnormal electrical events.     

GSK3 and β-catenin determines functional expression of sodium channels at the axon initial segment

Neuronal action potentials are generated through voltage-gated sodium channels, which are tethered by ankyrinG at the membrane of the axon initial segment (AIS). Despite the importance of the AIS in the control of neuronal excitability, the cellular and molecular mechanisms regulating sodium channel expression at the AIS remain elusive. Our results show that GSK3α/β and β-catenin phosphorylated by GSK3 (S33/37/T41) are localized at the AIS and are new components of this essential neuronal domain. Pharmacological inhibition of GSK3 or β-catenin knockdown with shRNAs decreased the levels of phosphorylated-β-catenin, ankyrinG, and voltage-gated sodium channels at the AIS, both “in vitro” and “in vivo”, therefore diminishing neuronal excitability as evaluated via sodium current amplitude and action potential number. Thus, our results suggest a mechanism for the modulation of neuronal excitability through the control of sodium channel density by GSK3 and β-catenin at the AIS.     

The CorA family: Structure and function revisited

The CorA family is a group of ion transporters that mediate transport of divalent metal ions across biological membranes. Metal ions are essential elements in most cellular processes and hence the concentrations of ions in cells and organelles must be kept at appropriate levels. Impairment of these systems is implied in a number of pathological conditions. CorA proteins are abundant among the prokaryotic organisms but homologues are present in both human and yeast. The activity of CorA proteins has generally been associated with the transport of magnesium ions but the members of the CorA family can also transport other ions such as cobalt and nickel. The structure of the CorA from Thermotoga maritima, which also was the first structure of a divalent cation transporter determined, has opened the possibilities for understanding the mechanisms behind the ion transport and also corrected a number of assumptions that have been made in the past.     

Anionic activation of human salivary amylase.

In all earlier studies on alpha-amylase, the influence of different ions were studied in phosphate buffer. The present report shows the effect of different ions individually with Tris and amino acid. Though it has been claimed recently that sodium ion is an activator of alpha-amylase, this study reconfirms that sodium ion does not activate human salivary amylase.     

Cardiac cellular electrophysiology: past and present

Summary The time-course of the cardiac action potential can be accounted for in terms of ionic currents crossing the cell membranes. Depolarizing current is carried by Na⁺ or Ca²⁺ entering the cells, repolarizing current by K⁺ leaving the cells. Membrane permeability for the passive movement of these ions is thought to be voltage-dependent as well as time-dependent. Net transfer of charge may also result from active transport, 2 Na⁺ out against 1 K⁺ in; or coupled exchange, 3 or 4 Na⁺ in against 1 Ca²⁺ out. This review follows the path by which present-day knowledge has been reached. It also gives a few examples to illustrate that electrophysiology has provided concepts useful to clinical cardiology.