Factors Influencing Adrenergic Mechanisms in the Heart: Satellite Symposium of the 28th International Congress of Physiological Science, Visegrá, Hungary, 1980

Szentiványi

EFFECTS OF ADRENALINE ON SOME ASPECTS OF ELECTRICAL AND MECHANICAL ACTIVITY IN THE FROG HEART

G. Vassort and R. Ventura-Clapier,     Laboratoire de Physiologie Cellulaire Cardiaque, FRA 51 INSERM, Bât 443, Université Paris-Sud, F-91405-Orsay, France

Publisher Summary

This chapter presents the effects of adrenaline that were investigated on three aspects of frog heart mechanical activity: increase in the slow current and related mechanical strength in normal medium or in metabolic-deficient solution and increase in the rate of relaxation of tension. These aspects are considered significant examples of three major topics in the excitation–contraction coupling of the heart muscle cells. The amplitude of tension is controlled not only by the free intracellular calcium (Ca) concentration but also by the energy-rich phosphates available at the myofilament sites; besides, the relaxation is dependent upon the rate of decrease in intracellular Ca following its uptake by the internal sites or extrusion through the cellular membrane. The experiments discussed in the chapter were performed under voltage clamp conditions on frog atrial trabeculae, 70–150 μm in diameter, by means of a double sucrose gap method with simultaneous recording of mechanical activity. Tension was measured in the test compartment with a transducer element lengthened by a thin lever that is 4 cm long and of 50 mg weight.

Myocardial behaviour is known to be altered by catecholamines. In the following the effects of adrenaline were investigated on three aspects of frog heart mechanical activity: increase in the slow current and related mechanical strength 1) in normal medium or 2) in metabolic-deficient solution and 3) increase in the rate of relaxation of tension. These aspects are considered significant examples of three major topics in the excitation-contraction coupling of the heart muscle cells: the amplitude of tension is controlled by the free intracellular Ca concentration but also by the energy-rich phosphates available at the myofilament sites; besides, the relaxation is dependent upon the rate of decrease in intracellular Ca following its uptake by the internal sites or extrusion through the cellular membrane.

The experiments were performed under voltage clamp conditions on frog (Rana esculenta) atrial trabeculae, 70–150 μm in diameter, by means of a double sucrose gap method with simultaneous recording of mechanical activity (Vassort and Rougier, 1972). Tension was measured in the test compartment with a transducer element (serie AE 800 from AME) lengthened by a thin lever (4 cm long; 50 mg weight).

Effect of adrenaline in normal medium

Two different inward currents are responsible for the development of the cardiac action potential. The Ca current may be distinguished from the initial Na current in several ways since the conductance mechanisms involved respond differently to drugs and since they have different dependences on potential and time. The initial current is completly inactivated by holding the membrane at −40 mV and is inhibited by tetrodotoxin (TTX); in both conditions a Ca current may be elicited by applied depolarizations. This latter current is inhibited by Mn or La ions and by verapamil or D 600. The rather slow inactivation time course suggests that the Ca conductance contributes to determine the duration of the action potential. The correlation between Ca current and tension is striking in the frog heart (Fig. 1). Tension increases rapidly when the Ca current first become appreciable. The peak tension occurs at the same potential as for peak Ca current and, when the membrane is further depolarized towards the Ca equilibrium potential both Ca current and tension decline. However when pulses longer than the 50 msec used here are applied, a second component of tension is elicited that is independent of Ca current. It increases with the amplitude of the depolarization and with the duration of the pulse up to about 1 second. This component of tension accounts for the fact that tension is maintained as long as depolarization is. This latter component was namedCytonic tension. The first one, dependent on the Ca current, is refered to as phasic tension" (Vassort and Rougier, 1972). These two components of tension are also evidenced in mammalian heart (Coraboeuf, 1974). Tonic tension was recently shown to depend on the Na-Ca exchange mechanism located at the cellular membrane (Horackova & Vassort, 1979).

Fig. 1 Effects of adrenaline (5 × 10−6M) on the slow Ca inward current and the phasic tension. A: for a given depolarizing step (95 mV-60 msec), adrenaline increases both the slow inward current (upper traces) and the mechanical activity (lower traces). The fast Na inward current is unchanged (arrows). B: peak tension and maximal inward current plotted against the depolarization applied to the membrane from its resting potential (Er) in Ringer solution (open symbols) and after the addition of adrenaline (closed symbols) (Vassort, 1973).

A further correlation between Ca current and phasic tension was provided by the effects of adrenaline. In addition to its chronotropic action in heart, adrenaline increased the force of contraction. Adrenaline has been found to increase the Ca current inflow (Vassort et al., 1969, Reuter, 1974). It also increased markedly the peak tension elicited by short depolarizations (Fig. 1). Both increase in current and tension occurred simultaneously with a half-time about 1 min. On the other hand, tonic tension did not seem significantly altered by adrenaline.

Effects of adrenaline during CN-poisoning

During CN poisoning, oxidative metabolism is impaired. This leads to a decrease in mechanical and electrical activities on which the ability of adrenaline to counteract the inhibition was tested.

On frog heart, cyanide produces an initial rapid decrease in peak tension followed by a decrease in action potential duration and amplitude (Fig. 2). Once tension was reduced to 20% of its initial value, adrenaline (10−6M) allowed only incomplete recovery of maximal tension whereas action potential amplitude was enhanced. Such weak effects of adrenaline has been also described for mammalian heart under hypoxia (Davidson et al., 1974).

Fig. 2 Effects of adrenaline (10−6M) in the presence of cyanide (3 × 10−3M) on the electrical and mechanical activities of frog heart. Upper traces: action potentials and tension recordings before and after 25 min in cyanide and a further 11 min in cyanide plus adrenaline taken from a representative experiment. Notice the recording was stopped for 14 min during cyanide application.

The same events were analysed in voltage clamp experiments. In cyanide, the two components of tension, phasic and tonic were decreased within 5 min prior to alteration in the Ca current. After 15 min both Ca current and tension were diminished to a steady value. When adrenaline (10−6M) was added, it produced a large and rapid increase in Ca current to about four times the peak current in Ringer solution while tension recovered nearly its initial value (Fig. 3B). The graph in Fig. 3A gives both the peak tension and peak current elicited by 60 mV–160 ms pulses (holding potential −40 mV) in order to differentiate the time courses of the rises in the slow inward current and tension induced by adrenaline (5 × 10−7M) in the presence of cyanide. It is noticeable that the time courses of both increases are very different. Peak inward current increased very rapidly with a half time of about 20 sec while the increase in tension appeared to be delayed and slow (half time: 90 sec). Such different time courses between electrical and mechanical activity following the addition of adrenaline has been shown in the hypodynamic frog heart (Niedergerke and Page, 1977).

Fig. 3 Analyses of current and tension under voltage clamp conditions. A The peak slow inward current (filled circles) and peak tension (filled triangles) elicited by 160 ms − 60 mV pulses (with a holding potential: Er + 30 mV) are plotted versus time to differenciate the time course of rises in current and tension after application of adrenaline (5 × 10−7M). The first two points show the amplitudes in normal Ringer. Zero time is taken as 7 min after the addition of cyanide. B Slow inward current and peak tension-voltage relationships illustrating the effects of adrenaline (5 × 10−7M) in the presence of cyanide (3 × 10−3M) (filled circles). Curves for normal Ringer (crosses) and after 20 min cyanide (filled squares) are