Sarcolemmal Ca Pump Research Articles

Lüttgau & Niedergerke; the classic study of calcium–sodium antagonism half a century on

Lüttgau & Niedergerke; the classic study of calcium–sodium antagonism half a century on

Excitation-Contraction Coupling and Cardiac Contractile Force

1. Major cellular structures involved in E-C coupling. 2. Myofilaments: The end effector of E-C Coupling. 3. Sources and sinks of activator calcium. 4. Cardiac action potentials and ion channels. 5. Ca influx via sarcolemmal Ca channels. 6. Na/Ca exchange and the sarcolemmal Ca-pump. 7. Sarcoplasmic reticulum Ca uptake, content and release. 8. Excitation-contraction coupling. 9. Control of cardiac contraction by SR and sarcolemmal Ca fluxes. 10. Cardiac inotropy and Ca mismanagement. References. Index.

Integrated Ca2+ management in cardiac myocytes.

Cardiac myocyte excitation-contraction coupling is complex. There are many systems involved that interact to form varied, but well-tuned, effects that are essential to contractile regulation. Nearly all of these systems are Ca-dependent, and Ca homeostasis within the myocyte is carefully controlled. Contractile activation results from Ca entry via Ca current, and Ca release from the sarcoplasmic reticulum (SR). Ca extrusion from the cytosol is controlled by Ca transport by (1) the Na-Ca exchanger, (2) the SR Ca-pump (which is balanced by a Ca leak out of the SR), and (3) slower systems (including Ca transport by mitochondria and the sarcolemmal Ca-pump). These systems interact to regulate the amount of Ca within the cell at rest, most of which is stored within the SR. The amount of Ca released from the SR depends nonlinearly upon SR [Ca], specifically the free SR [Ca] ([Ca](SR)). The relationship is particularly steep at high [Ca](SR), where spontaneous release can take place, resulting in electrical arrhythmias. In many models of heart failure, SR [Ca] is reduced, which may cause decreased Ca release and contractile dysfunction. In summary, the varied processes responsible for Ca regulation within the myocyte are critical to normal heart function, and disruption of the normal operation of these proteins can cause widely varied pathological effects, in large part due to dysfunctional Ca handling.

Relaxation in ferret ventricular myocytes: role of the sarcolemmal Ca ATPase.

In ferret ventricular myocytes the rate of intracellular Ca concentration [Ca]i decline and relaxation is remarkably fast (compared with rabbit and rat) under conditions where both the sarcoplasmic reticulum Ca uptake and Na/Ca exchange are inhibited. Here we explore the possibility that this rapid [Ca]i decline in ferret cells is attributable to the sarcolemmal Ca ATPase by using carboxyeosin (a potent inhibitor of the sarcolemmal Ca-ATPase). We compare the effects of carboxyeosin with those of elevated extracellular [Ca] ([Ca]o) (a thermodynamic approach to limit Ca transport by the sarcolemmal Ca ATPase). In rabbit cells, carboxyeosin and high [Ca]o slowed [Ca]i decline similarly and both virtually abolished [Ca]i decline when mitochondrial Ca uptake was also inhibited. In ferret cells, carboxyeosin treatment produced these same effects on [Ca]i decline, but high [Ca]o did not mimic them. Moreover, only in carboxyeosin-treated ferret cells did additional inhibition of mitochondrial Ca uptake nearly abolish [Ca]i decline. We conclude that, carboxyeosin loading can inhibit the sarcolemmal Ca-ATPase in intact myocytes; that this pump seems likely to be responsible for the much faster relaxation observed in ferret cells after block of SR Ca accumulation and Na/Ca exchange transport and that the sarcolemmal Ca pump apparently has different characteristics in rabbit and ferret ventricular myocytes.

Influence of extracellular potassium on energetics of resting heart muscle.

The effects of various extracellular K concentrations ([K]e) on energy expenditure and their relationship to ionic exchange mechanisms under quiescent conditions were investigated in the arterially perfused rat heart. The increase in [K]e (from 6 to 12, 24, or 50 mM K) leads to a rapid increase (results are given per gram dry weight) in resting energy expenditure (+5.9 +/- 0.9, +13.6 +/- 1.1, and +30.0 +/- 2.0 mW/g, respectively) followed by a slow decrease toward a new steady rate of heat production but higher (+2.8 +/- 0.7, +6.3 +/- 0.6, and +10.5 +/- 1.1 mW/g) than that observed under control conditions (21.1 +/- 0.7 mW/g). The increase in [K]e from 6 to 50 mM also induced an increase in K influx (calculated from 86Rb uptake and efflux experiments) of approximately 0.25 mumol.g-1.s-1. If this increased K influx is driven by the Na-K pump, an increase in steady resting heat production of approximately 10 mW/g would be expected. This represents 95% of the increase in steady heat production measured for 50 mM K intervention. The simultaneous increase in the cellular Ca flux (+0.1 mumol.g-1.min-1) can only explain (if driven by the sarcolemmal Ca pump) less than 1% of the steady increase in heat production. The work also shows that the initial, transitory increase in resting heat production induced by increasing [K]e is caffeine sensitive and may be at least partially attributable to a transitory enhanced activity of the sarcoplasmic reticulum.

Calcium and the heart: exchange at the tissue, cell, and organelle levels.

Cellular calcium (Ca) exchange in arterially perfused whole heart is markedly perfusion-limited. Therefore, a large fraction of exchangeable Ca recycles within the cell before exchanging with extracellular Ca. Ca enters the cell via sarcolemmal (SL) transient (T) and long-lasting (L) channels activated at more negative and less negative membrane potential, respectively. The larger L currents are controlled via phosphorylation and G protein interaction to provide Ca to induce Ca release from sarcoplasmic reticulum (SR) and for direct myofilament activation. Troponin C (TNC) binds 3 mol of Ca/mol. Only the low affinity site is responsible for activation of force and regulation of myofilament ATPase rate. This occurs through a shift in troponin I (TNI) with respect to actin induced by the TNC Ca binding. Relaxation depends on reduction of cytosolic (Ca) and occurs via 1) Ca pumping into the longitudinal SR modulated by phospholamban; 2) the recently cloned high-capacity electrogenic SL Na-Ca exchanger; and 3) the SL Ca pump under complex regulation including calmodulin control. Mitochondria transport Ca, but this transport is directed primarily to the regulation of various Ca-sensitive dehydrogenases so that oxidative metabolism can be adjusted to changes in energy demand. The regulation of Ca movements consumes about 25% of the cell's total energy output. Mitochondrial Ca exchanges with extracellular Ca most slowly (t 1/2 = 3.6 min), SR Ca quite rapidly (t 1/2's = 3 and 19 s), and an Na-Ca exchange-dependent compartment very rapidly (t 1/2 = 500 ms). After further description of Ca handling by the individual organelles, Ca movement is followed through the cell during the course of contraction, and the contribution of each organelle or compartment to overall cellular Ca exchange is defined.

O-148 - Cyclic-nucleotide regulation of sarcolemmal Ca pump in vascular smooth muscle cells

O-148 - Cyclic-nucleotide regulation of sarcolemmal Ca pump in vascular smooth muscle cells

Sodium/calcium exchange in ventricular muscle

Ventricular cells possess two Ca extrusion mechanisms, a Na/Ca exchange system and a Ca pump. Reversing the exchanger by extracellular Na removal causes [Na]i to decrease, and the cells take up mmolar quantities of calcium. Since [Ca]i shows only a marginal increase the calcium load must be buffered. The capacity of the SR is limited so the mitochondria probably buffer a large part of this load. However, when Ca uptake into the mitochondria is blocked, the gain in Ca is still mmolar and the increase in [Ca]i still marginal, suggesting an additional buffering site. Measurements of the Na/Ca stoichiometry on sarcolemmal vesicles gave a value of 3, but in ventricle values of around 2.5 or 3 are found. Reasons for this are discussed, as are the differences amongst the different methods of Ca measurement. The interaction of the sarcolemmal Ca pump and the exchanger are considered and it is suggested they could interact via [Na]i. At rest both systems could remove Ca from the cell but on a large perturbation the Na/Ca exchange would be the more important of the two.

Ryanodine and the calcium content of cardiac SR assessed by caffeine and rapid cooling contractures.

The ability of rabbit ventricular muscle sarcoplasmic reticulum (SR) to accumulate and retain Ca during Na-free perfusion was assessed using caffeine contractures and rapid cooling contractures (RCC). Muscles were exposed to a Na-free medium for 15 min, during which time a transient contracture developed and relaxed back to the resting tension level. Muscles were then either exposed to 20 mM caffeine or rapidly cooled to less than 5 degrees C, both of which produced large contractures. The magnitudes of those contractures are believed to be a relative index of SR Ca content. Reduction of extracellular [Ca] from 2 to 0.2 mM did not significantly alter the magnitude of either Na-free caffeine contractures or RCC. These are not the maximum contractures that can be obtained. The possibility that low extracellular [Ca] ([Ca]o) may increase passive Ca permeability is suggested in explanation of this effect. After equilibration with 100 nM ryanodine, both Na-free caffeine contractures and RCC are virtually eliminated. This suggests that even if the SR could accumulate Ca during the initial Na-free exposure in the presence of ryanodine, it could not retain that Ca in Na-free medium. It is proposed that the sarcolemmal Ca pump can extrude Ca from the cells at a rate sufficient to deplete the ryanodine-treated SR. When removal of Na was accompanied by increase of [K]o (to 12, 20, or 30 mM), the Na-free RCC were enhanced. Increasing [K]o during Na-free superfusion in the presence of ryanodine resulted in demonstrable RCC that were graded with [K] and completely suppressed by nifedipine.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of caffeine on energy output of rabbit heart muscle.

The effects of caffeine (1 mmol.l-1) on mechanical and energetic parameters in the arterially perfused interventricular rabbit septa were examined at various frequencies of stimulation. Even though 1 mmol-1 caffeine induced a negative inotropic effect only at stimulation rates higher than 0.33 Hz, relaxation was impaired at all frequencies tested. The ratio between maximum rate of relaxation and developed tension (-Tmax/T) was consistently lowered by caffeine, indicating a more marked effect on relaxation over contraction. In addition, while time-to-peak tension was unaffected by caffeine at the dose used, the last part of the relaxation (i.e., of the contractile event) was prolonged at all frequencies in the presence of the drug. Resting heat production (Hr) was increased in the presence of caffeine (1.6 +/- 0.6 mW.g-1). The ratios between active heat production and either developed tension (Ha/T) or tension time integral (Ha/TtI), increased at all frequencies examined (53.3 +/- 8.5 microJ.mN-1.g-1 and 68.2 +/- 9.9 microJ.mN-1.s-1.g-1, respectively), indicating a lowered economy of the contractile process. This is consistent with the lower ATP/Ca ratio reported for the sarcoreticular Ca pump (i.e., one ATP hydrolyzed/2 Ca transported) with respect to the sarcolemmal mechanisms such as Na-Ca exchanger or the sarcolemmal Ca pump, with an ATP/Ca ratio of 1 to 1. Thus, inhibition of the SR-Ca pump by caffeine would induce a higher rate of ATP hydrolysis with the consequent increase in the Ha/T ratio. As a result of the increase in both Ha/T ratio and Hr induced by caffeine, the ratio between total heat production and developed tension (Ht/T) also increased. Therefore, the contractile process appeared to be more efficient in the presence of an active SR, since it is energetically less costly to generate a given level of isometric tension.