Membrane Dysfunction and Abnormal Spontaneous Activity: A Study in Explanted Cardiac Cells

Author(s):  
Otto F. Schanne ◽  
Jihong Qu ◽  
George E. Haddad ◽  
Elena Ruiz-Petrich
1972 ◽  
Vol 50 (6) ◽  
pp. 523-532 ◽  
Author(s):  
O. F. Schanne

Beating cell clusters were obtained by trypsinization from hearts of newborn rats. Spontaneous activity ceased after several weeks while the cultures were still proliferating. Experiments were performed to identify the physiological determinant causing cessation of spontaneous activity. (a) Cell clusters having lost their spontaneous activity responded to extracellular stimulation. (b) Reduction of [K]o by 50% increased the number of beating cell clusters by 40%; doubling [K]o reduced the number of beating cell clusters by 49%. (c) Cell clusters which were in the process of losing their ability to contract spontaneously needed a progressively increasing temperature to induce spontaneous activity. These results suggest (1) that the pacemaker mechanism fails first when a cell cluster loses its spontaneous activity and (2) that shortly before the cluster fails to contract spontaneously, it requires more energy to maintain pacemaker activity because of possible structural membrane changes or changes in the enzyme pattern of the cells.


1997 ◽  
Vol 273 (2) ◽  
pp. H886-H892 ◽  
Author(s):  
R. A. Bassani ◽  
J. W. Bassani ◽  
S. L. Lipsius ◽  
D. M. Bers

Evidence has shown that the sarcoplasmic reticulum (SR) of cardiac cells releases Ca not only during excitation-contraction coupling but also during diastole, albeit at a much lower rate. This diastolic SR Ca release (leak) has also been implicated in the generation of spontaneous depolarization in latent atrial pacemaker cells of the cat right atrium. In the present work, we sought to measure Ca transients in pacemaker and nonpacemaker cells of the cat using the fluorescent Ca indicator indo 1. Atrial latent pacemaker cells develop a slow Ca transient when rested in the presence of both Na- and Ca-free solution and thapsigargin [used to inhibit Na/Ca exchange and SR Ca adenosinetriphosphatase (Ca-ATPase), respectively]. This increase in cytosolic Ca concentration ([Ca]i) is probably caused by the rate of SR Ca leak exceeding the capacity of the remaining Ca transport systems (e.g., sarcolemmal Ca-ATPase and mitochondrial Ca uptake). However, neither cat sinoatrial (SA) node cells nor myocytes from cat atrium or ventricle exhibited a similar increase in [Ca]i during the same protocol. This indicates that SR Ca leak in these cells occurred at a rate low enough to be within the capacity of the slow Ca transporters, as observed previously in rabbit ventricular myocytes. When atrial and ventricular myocytes were stimulated at higher frequencies, sufficient to markedly increase diastolic and systolic [Ca]i and approach Ca overload (and spontaneous activity), they responded to inhibition of SR Ca-ATPase and Na/Ca exchange with a slow Ca transient similar to that normally observed in atrial latent pacemaker cells. Furthermore, the SR Ca depletion by thapsigargin did not affect spontaneous activity of SA node cells, but it prevented or slowed pacemaker activity in the atrial latent pacemaker cells. These findings suggest that enhanced diastolic SR Ca efflux contributes significantly to the generation of spontaneous activity in atrial subsidiary pacemakers under normal conditions and in Ca-overloaded myocytes but not in SA node cells.


1975 ◽  
Vol 53 (6) ◽  
pp. 1209-1213
Author(s):  
O. F. Schanne ◽  
C. Rivard ◽  
G. Doyon

The spontaneous activity of cell clusters derived from ventricle cells of newborn rats was studied using a recording television microscope. The influence of varying concentrations of sodium, potassium, calcium, tetrodotoxin (TTX), and that of 2 mM MnCl2 was tested. The spontaneous activity of the cell clusters persisted in TTX but it was abolished by Mn. The beating rate increased when [Ca]0 and [Na]0 were changed from 0.3 mM to 3.0 mM and from 30 mM to 75 mM; it decreased with a change of [Na]0 from 75 mM to 142 mM. It is concluded that electrogenesis in these cells is determined by a slow inward current and that these cell clusters are comparable in their behavior to very young embryonic rat heart cells or cells of the rabbit sinoauricular node.


Author(s):  
W.G. Wier

A fundamentally new understanding of cardiac excitation-contraction (E-C) coupling is being developed from recent experimental work using confocal microscopy of single isolated heart cells. In particular, the transient change in intracellular free calcium ion concentration ([Ca2+]i transient) that activates muscle contraction is now viewed as resulting from the spatial and temporal summation of small (∼ 8 μm3), subcellular, stereotyped ‘local [Ca2+]i-transients' or, as they have been called, ‘calcium sparks'. This new understanding may be called ‘local control of E-C coupling'. The relevance to normal heart cell function of ‘local control, theory and the recent confocal data on spontaneous Ca2+ ‘sparks', and on electrically evoked local [Ca2+]i-transients has been unknown however, because the previous studies were all conducted on slack, internally perfused, single, enzymatically dissociated cardiac cells, at room temperature, usually with Cs+ replacing K+, and often in the presence of Ca2-channel blockers. The present work was undertaken to establish whether or not the concepts derived from these studies are in fact relevant to normal cardiac tissue under physiological conditions, by attempting to record local [Ca2+]i-transients, sparks (and Ca2+ waves) in intact, multi-cellular cardiac tissue.


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