Beta-adrenergic inhibition of cardiac sodium channels by dual G-protein pathways

The signaling pathways by which beta-adrenergic agonists modulate voltage-dependent cardiac sodium currents are unknown, although it is likely that adenosine 3'5'-monophosphate (cAMP) is involved. Single-channel and whole-cell sodium currents were measured in cardiac myocytes and the signal transducing G protein Gs was found to couple beta-adrenergic receptors to sodium channels by both cytoplasmic (indirect) and membrane-delimited (direct) pathways. Hence, Gs can act on at least three effectors in the heart: sodium channels, calcium channels, and adenylyl cyclase. The effect on sodium currents was inhibitory and was enhanced by membrane depolarization. During myocardial ischemia the sodium currents of depolarized cells may be further inhibited by the accompanying increase in catecholamine levels.     

Sodium current-induced release of calcium from cardiac sarcoplasmic reticulum

The role of sodium-calcium exchange at the sarcolemma in the release of calcium from cardiac sarcoplasmic reticulum was investigated in voltage-clamped, isolated cardiac myocytes. In the absence of calcium entry through voltage-dependent calcium channels, membrane depolarization elicited release of calcium from ryanodine-sensitive internal stores. This process was dependent on sodium entry through tetrodotoxin-sensitive sodium channels. Calcium release under these conditions was also dependent on extracellular calcium concentration, suggesting a calcium-induced trigger release mechanism that involves calcium entry into the cell by sodium-calcium exchange. This sodium current-induced calcium release mechanism may explain, in part, the positive inotropic effects of cardiac glycosides and the negative inotropic effects of a variety of antiarrhythmic drugs that interact with cardiac sodium channels. In response to a transient rise of intracellular sodium, sodium-calcium exchange may promote calcium entry into cardiac cells and trigger sarcoplasmic calcium release during physiologic action potentials.     

Functional differences between two classes of sodium channels in developing rat skeletal muscle

Excitability is generated in developing skeletal muscle by the incorporation of sodium-selective ion channels into the surface membrane. Whole-cell and patch voltage-clamp recording from myotubes and their embryologic precursors, myoblasts, indicated that voltage-activated sodium current in myoblasts was more resistant to block by tetrodotoxin (TTX) than that in myotubes. Single-channel recording from both cell types showed two classes of sodium channels. One class had a lower single-channel conductance, activated at more hyperpolarized voltages, and was more resistant to TTX than the other. The proportion of TTX-resistant to TTX-sensitive sodium channels was higher in myoblasts than in myotubes. Thus, the difference in TTX sensitivity between myoblasts and myotubes can be explained by a difference in the proportion of the two classes of sodium channels. In addition, the lower conductance of TTX-resistant channels provides insight into the relationship between the TTX binding site and the external mouth of the sodium channel.     

Calcium current and activation of contraction in ventricular myocardial fibers

In thin bundles of dog ventricular myocardium, a slow inward current (distinct from the sodium inward current) could be recorded under voltageclamp conditions. This inward current was influenced by changes in external calcium concentration, but it was not dependent on external sodium concentration. Therefore, this current which contributes an appreciable amount of charge transfer during the plateau of the action potential, is carried by calcium ions. In sodium-free solution, the flow of calcium ions into the fiber is directly related to activation of contraction. In sodium-containing solution, however, calcium inward current serves primarily to fill up some intracellular stores from which calcium can be released by moderate depolarization.     

The relationship between charge movements associated with ICa and INa-Ca in cardiac myocytes

Ventricular myocytes exhibit a nifedipine-sensitive inward calcium current (ICa) and contracture when they are voltage clamped from -40 to 0 millivolt in the presence of caffeine and in the absence of extracellular sodium. However, upon repolarization they fail to relax because neither the sarcoplasmic reticulum nor the sodium-calcium exchange can reduce intracellular calcium. Sudden application of extracellular sodium during the contracture (but after repolarization) causes immediate relaxation and activates a transient inward sodium-calcium exchange current (INa-Ca), whose peak slightly precedes mechanical relaxation. The total charge carried by the nifedipine-sensitive ICa is twice the total charge carried by the transient inward INa-Ca. Assuming an exchange stoichiometry of three sodium to one calcium, these results indicate that all the calcium entering the cell during the initial depolarization is extruded by the sodium-calcium exchange.     

Sodium current in ventricular myocardial fibers

Membrane currents were measured in thin bundles of dog ventricular myocardium under voltage-clamp conditions. A rather large initial inward current which had an equilibrium potential at about + 55 millivolts could be recorded. When the external sodium concentration was reduced, the equilibrium potential for this current was shifted by the amount predicted theoretically for a current carried solely by sodium ions. The size of the sodium inward current (I(Na)) was largely dependent on the preceding membrane potential. The I(Na) was completely inactivated if the membrane potential was as low as -45 millivolts. Sodium ions are the main carrier of charge during the rapid depolarization phase of the action potential.     

Atrionatriuretic peptide transforms cardiac sodium channels into calcium-conducting channels

The atrionatriuretic peptide (ANP) is released from atrial cells in response to increased extracellular fluid volume and reduces sodium absorption by the kidney, thus reducing the blood volume. In this report, ANP suppressed the calcium and sodium currents in rat and guinea pig ventricular myocytes. The suppression of sodium current was caused by enhanced permeability of the sodium channel to calcium without significant changes in the kinetics or the tetrodotoxin sensitivity of the channel. Thus, ANP may regulate the sodium channel by altering its cationic selectivity site to calcium, thereby repressing the sodium current. The suppression of sodium and calcium channels and the resultant depressed excitability of the atrial cells may help to regulate ANP secretion.     

Changes in sodium channel gating produced by point mutations in a cytoplasmic linker

Voltage-gated sodium channels are transmembrane proteins of approximately 2000 amino acids and consist of four homologous domains (I through IV). In current topographical models, domains III and IV are linked by a highly conserved cytoplasmic sequence of amino acids. Disruptions of the III-IV linker by cleavage or antibody binding slow inactivation, the depolarization-induced closed state characteristic of sodium channels. This linker might be the positively charged "ball" that is thought to cause inactivation by occluding the open channel. Therefore, groups of two or three contiguous lysines were neutralized or a glutamate was substituted for an arginine in the III-IV linker of type III rat brain sodium channels. In all cases, inactivation occurred more rapidly rather than more slowly, contrary to predictions. Furthermore, activation was delayed in the arginine to glutamate mutation. Hence, the III-IV linker does not simply act as a charged blocker of the channel but instead influences all aspects of sodium channel gating.     

Hyperkalemic periodic paralysis and the adult muscle sodium channel alpha-subunit gene

Hyperkalemic periodic paralysis (HYPP) is an autosomal dominant disorder characterized by episodes of muscle weakness due to depolarization of the muscle cell membrane associated with elevated serum potassium. Electrophysiological studies have implicated the adult muscle sodium channel. Here, portions of the adult muscle sodium channel alpha-subunit gene were cloned and mapped near the human growth hormone locus (GH1) on chromosome 17. In a large pedigree displaying HYPP with myotonia, these two loci showed tight linkage to the genetic defect with no recombinants detected. Thus, it is likely that the sodium channel alpha-subunit gene contains the HYPP mutation.     

Ca2+ and Ca2+-activated K+ currents in mammalian gastric smooth muscle cells

Inward movement of calcium through voltage-dependent channels in muscle is thought to initiate the action potential and trigger contraction. Calcium-activated potassium channels carry large outward potassium currents that may be responsible for membrane repolarization. Calcium and calcium-activated potassium currents were identified in enzymatically isolated mammalian gastric myocytes. These currents were blocked by cadmium and nifedipine but were not substantially affected by diltiazem or D600. No evidence for a tetrodotoxin-sensitive sodium current or an inwardly rectifying potassium current was found.     

Functional properties of rat brain sodium channels expressed in a somatic cell line

Transfection of Chinese hamster ovary cells with complementary DNA encoding the RIIA sodium channel alpha subunit from rat brain led to expression of functional sodium channels with the rapid, voltage-dependent activation and inactivation characteristic of sodium channels in brain neurons. The sodium currents mediated by these transfected channels were inhibited by tetrodotoxin, persistently activated by veratridine, and prolonged by Leiurus alpha-scorpion toxin, indicating that neurotoxin receptor sites 1 through 3 were present in functional form. The RIIA sodium channel alpha subunit cDNA alone is sufficient for stable expression of functional sodium channels with the expected kinetic and pharmacological properties in mammalian somatic cells.     

The calcium store sensor, STIM1, reciprocally controls Orai and CaV1.2 channels

Calcium signals, pivotal in controlling cell function, can be generated by calcium entry channels activated by plasma membrane depolarization or depletion of internal calcium stores. We reveal a regulatory link between these two channel subtypes mediated by the ubiquitous calcium-sensing STIM proteins. STIM1 activation by store depletion or mutational modification strongly suppresses voltage-operated calcium (Ca(V)1.2) channels while activating store-operated Orai channels. Both actions are mediated by the short STIM-Orai activating region (SOAR) of STIM1. STIM1 interacts with Ca(V)1.2 channels and localizes within discrete endoplasmic reticulum/plasma membrane junctions containing both Ca(V)1.2 and Orai1 channels. Hence, STIM1 interacts with and reciprocally controls two major calcium channels hitherto thought to operate independently. Such coordinated control of the widely expressed Ca(V)1.2 and Orai channels has major implications for Ca(2+) signal generation in excitable and nonexcitable cells.     

BKCa-Cav channel complexes mediate rapid and localized Ca2+-activated K+ signaling

Large-conductance calcium- and voltage-activated potassium channels (BKCa) are dually activated by membrane depolarization and elevation of cytosolic calcium ions (Ca2+). Under normal cellular conditions, BKCa channel activation requires Ca2+ concentrations that typically occur in close proximity to Ca2+ sources. We show that BKCa channels affinity-purified from rat brain are assembled into macromolecular complexes with the voltage-gated calcium channels Cav1.2 (L-type), Cav2.1 (P/Q-type), and Cav2.2 (N-type). Heterologously expressed BKCa-Cav complexes reconstitute a functional "Ca2+ nanodomain" where Ca2+ influx through the Cav channel activates BKCa in the physiological voltage range with submillisecond kinetics. Complex formation with distinct Cav channels enables BKCa-mediated membrane hyperpolarization that controls neuronal firing pattern and release of hormones and transmitters in the central nervous system.     

急性心肌缺血大鼠左心室流出道自发性慢反应电位去极离子流变化分析

目的探讨急性心肌缺血大鼠左室流出道自发性慢反应电位去极离子流的变化。方法常规玻璃微电极细胞内方法观测急性心肌缺血大鼠的离体心脏最大舒张电位(MDP)、0相除极幅度(APA)、0相最大除极速度(Vmax)、4相自动除极速度(VDD)、复极50%(APD50)和90%(APD90)的时间以及自发放电频率(RPF)。结果与给药前相比①1.2mmol.L-1河豚毒(TTX)使APA和Vmax有所减小(P<0.05),VDD和RPF明显减慢(P<0.01);②1.0μmol.L-1维拉帕米(VER)可使该慢电位的APA、Vmax、VDD明显减小,RPF减慢(P<0.01);APD、APD90延长(P<0.05);③2mmol.L-1 4-氨基吡啶(4-AP)使该慢电位的MDP的绝对值、APA、Vmax减小,VDD和RPF加快(P<0.01);④1.5mmol.L-1 CsCl作用6min时,VDD和RPF明显降低(P<0.05),10min时恢复。结论①左心室流出道的自发慢电位0相主要去极离子流除Ca2+内流外,还有少量Na+内流。②4相去极离子流中,除Ca2+、Na+的内流和Ik衰减外,If电流可能也起部分作用。     

Coding channels in the taste system of the rat

Basic taste qualities are thought to be perceived independently, yet discrete neural coding channels have not been demonstrated in the central nervous system. The response profiles of taste cells in the nucleus tractus solitarius (NTS) of the rat were categorized into four groups, and the effects of amiloride, a passive sodium channel blocker, on each were determined. NTS neurons that responded specifically to sodium chloride (NaCl) or to NaCl and sugars were suppressed by amiloride; those broadly sensitive to salts, acids, and bitter stimuli were unaffected. Moreover, the response profile evoked by NaCl lost its distinctiveness after treatment with amiloride, becoming similar to those evoked by acids and quinine. Receptors that respond to sodium must relay their information through independent coding channels to identifiable subgroups of NTS neurons, the activity of which is responsible for the perception of saltiness.     

Intracellular acidosis enhances the excitability of working muscle

Intracellular acidification of skeletal muscles is commonly thought to contribute to muscle fatigue. However, intracellular acidosis also acts to preserve muscle excitability when muscles become depolarized, which occurs with working muscles. Here, we show that this process may be mediated by decreased chloride permeability, which enables action potentials to still be propagated along the internal network of tubules in a muscle fiber (the T system) despite muscle depolarization. These results implicate chloride ion channels in muscle function and emphasize that intracellular acidosis of muscle has protective effects during muscle fatigue.     

Gamma-aminobutyric acid: role in primary afferent depolarization

The effects of putative transmitters on the primary afferent terminals were studied in the magnesium-treated, isolated spinal cord of the frog. Gamma-aminobutyric acid and glutamic acid reversibly depolarized primary afferent terminals and increased their excitability, whereas glycine produced weak and variable effects. Bicuculline and picrotoxin, which reduce primary afferent depolarization, reversibly antagonized the gamma-aminobutyric acid-mediated responses but had little effect on those produced by either glutamic acid or glycine. The glutamic acid- and the gamma-aminobutyric acid-induced depolarizations remained in the absence of external chloride but disappeared in the absence of external sodium. These results support the hypotheses that gamma-aminobutyric acid is the transmitter mediating the synaptic depolarization of primary afferent terminals and that sodium is the predominant ion involved.     

Membrane-Potential Alteration During K＋ Uptake of Different Salt-Tolerant Wheat Varieties

K＋ is the most abundant cation in plant cells and plays an important role in many ways.K＋ uptake of plant has respect to its salt resistant capacity.There are two categories of channel transportation for plants to uptake K＋,one is through K＋ channels and the other is through nonselective cation channels（NSCCs）.The transmembrane localization of K＋ may change membrane potential（MP）.In this paper,three wheat varieties with different salt tolerance were selected and the MP was measured by microelectrode during K＋ uptake.The results showed that the effects of K＋ uptake on MP through K＋ channels or NSCCs were distinct.K＋ influx through K＋ channels led to MP hyperpolarization,while K＋ influx through NSCCs resulted in depolarization.Diverse MP alteration of wheat varieties with different salt tolerance was mainly due to NSCCs-mediated K＋ uptake.Compared with the salt-tolerant wheat,the MP hyperpolarization during K＋ uptake of saltsensitive wheat was much more evident,probably because of the cation outflux through NSCCs during this process.     

Direct evidence for the cathodic depolarization theory of bacterial corrosion

Cathodic depolarization of mild steel by Desulfovibrio desulfuricans was demonstrated with benzyl viologen used as an electron acceptor. Direct measurement of the cathodic depolarization current indicated a maximum current density of 1 microampere per square centimeter. Aluminum alloys were also cathodically depolarized by the organism.     

Membrane currents examined under voltage clamp in cultured neuroblastoma cells

Examination of ionic membrane currents in a voltage-clamped neuronal cell line derived from the mouse C1300 neuroblastoma disclosed four kinetically different components: sodium, potassium, calcium, and leakage current. The kinetics, voltage dependence, and pharmacological properties of the sodium and potassium currents qualitatively resemble those of the corresponding currents in squid giant axon and frog myelinated nerve fiber, suggesting that the molecular structures of the sodium and potassium channels in neuroblastoma are similar to those of the non-mammalian preparations.