The Na+/Ca2+ exchanger, an ion transport protein, is expressed in the plasma membrane (PM) of virtually all animal cells. It extrudes Ca2+ in parallel with the PM ATP-driven Ca2+ pump. As a reversible transporter, it also mediates Ca2+ entry in parallel with various ion channels. The energy for net Ca2+ transport by the Na+/Ca2+ exchanger and its direction depend on the Na+, Ca2+, and K+ gradients across the PM, the membrane potential, and the transport stoichiometry. In most cells, three Na+ are exchanged for one Ca2+. In vertebrate photoreceptors, some neurons, and certain other cells, K+ is transported in the same direction as Ca2+, with a coupling ratio of four Na+ to one Ca2+ plus one K+. The exchanger kinetics are affected by nontransported Ca2+, Na+, protons, ATP, and diverse other modulators. Five genes that code for the exchangers have been identified in mammals: three in the Na+/Ca2+ exchanger family (NCX1, NCX2, and NCX3) and two in the Na+/Ca2+ plus K+ family (NCKX1 and NCKX2). Genes homologous to NCX1 have been identified in frog, squid, lobster, and Drosophila. In mammals, alternatively spliced variants of NCX1 have been identified; dominant expression of these variants is cell type specific, which suggests that the variations are involved in targeting and/or functional differences. In cardiac myocytes, and probably other cell types, the exchanger serves a housekeeping role by maintaining a low intracellular Ca2+ concentration; its possible role in cardiac excitation-contraction coupling is controversial. Cellular increases in Na+ concentration lead to increases in Ca2+ concentration mediated by the Na+/Ca2+ exchanger; this is important in the therapeutic action of cardiotonic steroids like digitalis. Similarly, alterations of Na+ and Ca2+ apparently modulate basolateral K+ conductance in some epithelia, signaling in some special sense organs (e.g., photoreceptors and olfactory receptors) and Ca2+-dependent secretion in neurons and in many secretory cells. The juxtaposition of PM and sarco(endo)plasmic reticulum membranes may permit the PM Na+/Ca2+ exchanger to regulate sarco(endo)plasmic reticulum Ca2+ stores and influence cellular Ca2+ signaling.

https://www.physiology.org/doi/pdf/10.1152/physrev.1999.79.3.763

Sodium/Calcium Exchange: Its Physiological Implications

The essential connection of the basic physiological processes of excitation and contraction with transmembrane movements of Na, K, and Ca ions probably origi­ nates from an early stage of cellular evolution. Despite innumerable modifications the fundamental processes that developed in the ocean have not undergone major changes during the course of development of higher forms of animal life. Thus in a wide variety of excitable cells the transmembrane exchange of the monovalent marine cations Na and K can be considered the substantial basis of bioelectric membrane activity, whereas Ca ions are required as mediators when, by this superfi­ cial process, intracellular reactions such as muscular contraction, glandular secre­ tion, or liberation of transmitter substances are initiated ( 1, 2). Ca ions can exert this messenger function either in a primitive way, by penetrating into the intracellu­ lar space across the depolarized cell membrane or, at a more advanced stage of evolution, by being released from intracellulariy located endoplasmic stores. As to contractile tissues, the development of large endoplasmic Ca pools is most obvious in skeletal muscle, whereas myocardial fibers and, particularly, smooth muscle cells are less specialized in this respect. The natural consequences are as follows:

Specific Pharmacology of Calcium in Myocardium, Cardiac Pacemakers, and Vascular Smooth Muscle

A computer model of atrial fibrillation.

The development of new CRISPR-Cas genome editing tools continues to drive major advances in the life sciences. Four classes of CRISPR-Cas-derived genome editing agents-nucleases, base editors, transposases/recombinases and prime editors-are currently available for modifying genomes in experimental systems. Some of these agents have also moved rapidly into the clinic. Each tool comes with its own capabilities and limitations, and major efforts have broadened their editing capabilities, expanded their targeting scope and improved editing specificity. We analyze key considerations when choosing genome editing agents and identify opportunities for future improvements and applications in basic research and therapeutics.

Genome editing with CRISPR–Cas nucleases, base editors, transposases and prime editors

The arrhythmogenic role of increased dispersion of repolarization (dispersion) was studied in 23 open-chest dogs using six simultaneously recorded monophasic action potentials (MAPs) from the ventricular surface and programmed ventricular premature stimulation (VPS). Increased dispersion was induced by generalized hypothermia (29 degrees C) and regional warm blood (38-43 degrees C) perfusion through a coronary artery branch. Hypothermia and regional warm blood perfusion increased maximum dispersion from 13 +/- 10 to 111 +/- 16 msec (p less than 0.001), predominantly because of the increased MAP duration difference (10 +/- 15 vs 97 +/- 16 msec, p less than 0.001). The maximal difference between activation times was not significantly changed, but the QRS duration increased from 47 +/- 6 to 52 +/- 7 msec (p less than 0.01). Ventricular arrhythmia did not occur spontaneously but was induced by a single VPS in all 23 dogs during hypothermia and regional warm blood perfusion when dispersion reached a critical magnitude. The critical magnitude of dispersion required to induce ventricular arrhythmia was documented in 16 dogs by stepwise increments or decrements of dispersion. In four dogs, an increase in atrial pacing rate of 24 beats/min prevented induction of ventricular arrhythmia by decreasing dispersion from a critical magnitude of 103 +/- 5 msec to a nonarrhythmogenic value of 86 +/- 9 msec (p less than 0.05). In six dogs, we compared the stimulation site-dependent effects of VPS applied in the region with short and long MAPs. In all dogs, ventricular arrhythmia was inducible only by VPS from the region with a short MAP. Premature impulses from this region propagated more slowly than those from the region with a long MAP. Our results show that the large dispersion of repolarization facilitates the development of a conduction delay necessary to induce sustained arrhythmia by an early premature stimulus applied at the site with a short MAP.

Characteristics and possible mechanism of ventricular arrhythmia dependent on the dispersion of action potential durations.

Transmembrane activity was recorded from canine false tendons bathed with Tyrode's solution at 37°C. Stimulus patterns provided a 3-second pause after every ten beats. Acetylstrophanthidin was infused at concentrations up to 2x10-7 g/ml. One or two transient depolarizations (TDs) followed the last driven response of each series. The appearance of TDs was associated with depression of normal phase-4 depolarization. The peak of the earliest TD (TD-1) occurred at an interval approximately equal to the basic cycle length. The later TD (TD-2) occurred at about twice the basic cycle length. Coupling intervals were determined primarily by the last cycle length. The amplitude of TD-1 was maximal when the basic cycle length was 600 msec, but TD-2 continued to increase as the basic cycle length diminished further. The amplitude of both TDs increased with the number of beats in the train. Either or both could reach threshold and induce single extrasystoles or trains of extrasystoles. TDs could be induced to reach th...

A Cellular Mechanism for the Generation of Ventricular Arrhythmias by Acetylstrophanthidin

The intranodal propagation of atrial and ventricular echoes in in-vitro preparations of the rabbit heart was traced from microelectrode records at many puncture sites. Composite records indicate that the upper part of the node can be functionally and spatially dissociated into two pathways (α and β), which communicate with a final common pathway (FCP) about halfway between the atrium and the His bundle. Atrial echoes, induced by premature atrial stimulation, traverse the α pathway and return to the atrium from the middle part of the node over the β route; echoes can occur without propagation of the premature response to the ventricle. Ventricular echoes, induced by premature stimulation of the His bundle, pass from the FCP to the atrium over the α route, and back to the FCP over the β route. Repetitive reciprocation, in the nature of an intranodal circus movement, was commonly observed. The results completely substantiate previous conclusions based on indirect observations in the dog heart.

Demonstration of a Dual A-V Nodal Conduction System in the Isolated Rabbit Heart

Excessive delay and configurational change of ventricular responses to atrial premature beats, previously attributed to dissociation within the A-V node, were shown to be due to functional block of the right bundle branch. It was found that the refractory period (RP) of the right bundle branch often exceeded the functional refractory period (FRP) of the A-V node at slow heart rates, permitting an early premature atrial response to reach the ventricles before recovery of the right bundle. Vagal stimulation and rapid driving frequencies, by delaying the intranodal transit of the premature response, prevented the exposure of bundle branch block. Epinephrine in low doses sometimes facilitated its occurrence.

Premature responses initiated in the His bundle in open heart preparations were commonly blocked in the right bundle under conditions in which atrial premature responses were normally propagated to the ventricles. Evidence was obtained in such preparations that the portion of the right bundle system beyond the site of the block was activated retrogradely from the left side. Under certain conditions the retrograde activation process returned to the His bundle and the atrium as an echo.

Aberrant A-V Impulse Propagation in the Dog Heart: A Study of Functional Bundle Branch Block

Interactions between Purkinje fibers and ventricular muscle were studied in canine Purkinje-papillary muscle preparations. The change in duration of action potentials across the junctions between Purkinje fibers and papillary muscle was continuously graded, not abrupt. The transmembrane action potentials were longest in the false tendons and progressively shorter in more peripheral fibers. The responses of terminal Purkinje fibers and of neighboring muscle cells differed little in duration. The shortest action potentials were found in muscle fibers located in regions devoid of specialized tissue (tip of papillary muscle). These results suggest that the intercellular connections, including those at the junctions, offer relatively low resistance to current flow. During repolarization the current flowing between neighboring elements with intrinsically different repolarization times should therefore minimize the disparity in action potential durations on the two sides of the junctional site; a continuously graded change in duration would result. Because of this continuous gradation, premature ventricular responses initiated at the tip of the papillary muscle could be blocked, depending on the degree of prematurity, at various levels in muscle fibers functionally close to terminal Purkinje fibers or within the Purkinje system.

Interaction of Transmembrane Potentials in Canine Purkinje Fibers and at Purkinje Fiber-Muscle Junctions

In canine Purkinje fiber-papillary muscle preparations it was shown directly that electrotonic spread can take place across the junctions between Purkinje fibers and ordinary muscle fibers (P-M junctions). The P-M delay recorded during orthodromic propagation varied considerably within any given preparation. Moderate increases in the external concentration of K + (up to 6 mM) consistently decreased the P-M delay; when the concentration of external K + was increased to 8 mM the P-M delay increased. With larger concentrations of K + (10 to 11 mM), total conduction block from the terminal Purkinje fibers to the muscle occurred; under the same conditions antidromic propagation from muscle to Purkinje fiber was still possible. The results can be explained satisfactorily by assuming that propagation across the P-M junction is electrical and that the geometry of the functional syncytium changes progressively from a cable-like system at the level of the terminal Purkinje fibers to a two- or three-dimensional irregular syncytium at the bulk of the myocardial mass.

Carlos Mendez

Papers

A Cellular Mechanism for the Generation of Ventricular Arrhythmias by Acetylstrophanthidin

Demonstration of a Dual A-V Nodal Conduction System in the Isolated Rabbit Heart

Aberrant A-V Impulse Propagation in the Dog Heart: A Study of Functional Bundle Branch Block

Interaction of Transmembrane Potentials in Canine Purkinje Fibers and at Purkinje Fiber-Muscle Junctions

Propagation of Impulses across the Purkinje Fiber-Muscle Junctions in the Dog Heart