Quiescent Papillary Muscles Research Articles

Effects of aging on the work output and efficiency of rat papillary muscle.

This study aimed to investigate the effect of aging on the work output and efficiency of rat papillary muscle. The mechanical and energetic properties of left ventricular papillary muscle preparations isolated from 6-, 15-, and 27- to 32-month-old Sprague-Dawley rats were measured in myothermic experiments at 27 degrees C at a stimulus frequency of 0.167 Hz. We found that the basal metabolism measured in quiescent papillary muscles was significantly reduced in the 27- to 32-month-old group (4.9 mWg(-1) compared to 7.7 and 7.0 mWg(-1) in the 6- and 15-month groups). In isotonic experiments, the work output (at a range of afterloads) was significantly depressed for the 27- to 32-month group being only 52% of the work output of the 6-month group. This outcome was due to a decrease in both the extent of muscle shortening only, 66% of 6- and 15-month data, and in the maximum force developed. The reduced work was accompanied by a parallel decrease in energy consumption (enthalpy) and hence, the net mechanical efficiency (work/active enthalpyx100%) was not altered. A force-length- area (FLA) analysis was applied to the isotonic data and an energy: FLA regression line was obtained for each preparation. We found that there were no significant differences in either the intercept or slope of the energy: FLA relation with age. Contractile efficiency (39+/-3%) in the 27- to 32-month group was not significantly different to that found in the 6-month (43+/-4%) or 15-month (40+/-3% group). There are no changes in the mechanical performance or efficiency of cardiac muscle from young (6-month-old) or adult (15-month-old) rats but in the aged and senescent rats (27-32-month-old) there is a pronounced decline in stress development and shortening ability leading to a fall in work output. Mechanical and contractile efficiency however remain unchanged in old age and the data resembles that obtained in pressure overload hypertrophy.

Beta 1 adrenoceptor mediated decrease in pHi in quiescent ventricular myocardium.

The aims were to examine the effect of beta adrenergic stimulation on the intracellular pH (pHi) and to compare it with that of alpha adrenergic stimulation in ventricular myocardium. Using conventional and ion selective electrodes membrane potential and pHi were measured simultaneously in quiescent papillary muscles of guinea pigs in HEPES or bicarbonate buffered solution. Isoprenaline and propranolol (1 microM) plus phenylephrine (30 microM) were used to stimulate beta and alpha adrenoceptors, respectively. In order to evaluate underlying mechanism(s) of beta adrenoceptor mediated pHi change, effects of Na(+)-H+ exchange, Cl(-)-HCO3- exchange, Na(+)-HCO3- symport, and glycolysis blockers on the pHi change were examined. Isoprenaline (1 microM) produced a decrease in pHi of 0.08(SEM 0.01) pH units and a transient depolarisation of the resting membrane. The isoprenaline induced intracellular acidosis was blocked by the beta 1 blocker atenolol (10 microM) but not by the beta 2 blocker ICI 118,551 (0.1 microM). Forskolin also produced a decrease in pHi of 0.06(0.03) pH units. In contrast, alpha adrenergic stimulation produced an increase in pHi, which was abolished by 1 mM amiloride, an Na(+)-H+ exchange blocker. In the presence of amiloride, the isoprenaline induced decrease in pHi was rather enhanced. 4,4'-Diisothiocyanostilbene-2,2'-disulphonic acid (DIDS, 1 mM), a blocker of Cl(-)-HCO3- exchange and the Na(+)-HCO3- symport system, failed to affect the isoprenaline induced pHi decrease in bicarbonate buffered solution. However, pretreatment with 2-deoxyglucose or iodoacetic acid abolished the isoprenaline induced pHi decrease. beta 1 Adrenoceptor stimulation causes intracellular acidosis via the enhanced glycolysis, and the Na(+)-H+ exchange system appears to play a compensatory role. The beta 1 adrenoceptor mediated intracellular acidosis may modulate inotropic response to adrenergic stimulation in ventricular myocardium.

Intracellular Na activity measurements in the control and hypertrophied heart of the ferret: an ion-sensitive micro-electrode study

Because of the role of intracellular Na on cardiac contractility and of the depressed isometric contractile response of the hypertrophied myocardium, the effects of pressure overload on the intracellular Na activity (aiNa) have been investigated in papillary muscles isolated from the ferret right ventricle. In animals subjected to pulmonary artery clipped for 1-2 months, right ventricle-to-body weight ratio was increased by about 39% in comparison with the control group. aiNa was measured in quiescent papillary muscles, by means of Na-sensitive micro-electrodes, at room temperature (19-22 degrees C). aiNa values were, in the control ventricular cells, 7.8 +/- 1.1 mM (mean +/- SD; n = 20) and in the hypertrophied ones, 8.0 +/- 1.2 mM (n = 49). During superfusion by medium with a reduced extracellular Na concentration ([Na]0), aiNa declined in control and pressure-overloaded muscles to similar steady-state levels at a given [Na]0. aiNa fall was mono-exponential and was characterized by a smaller time constant in the hypertrophied group upon total withdrawal of Na0 (control 209 +/- 19 s, n = 4; hypertrophied 128 +/- 42 s, n = 6). In the absence of external K, aiNa increased to levels that were not significantly different between both groups. It was concluded that, in quiescent preparations, steady-state aiNa was not modified by the hypertrophic process. However, pressure overload induced a modification of aiNa regulation by a possible alteration of the sarcolemmal Na/Ca exchange, although other mechanisms, such as mitochondrial Ca transport, could be involved in the differential response to Na0 removal.

Intracellular sodium activity in papillary muscle from diabetic rat hearts

In quiescent papillary muscles isolated from hearts of rats with streptozotocin-induced diabetes resting intracellular sodium activity (aiNa) was about 56% greater than in muscles from controls. An intracellular acid load induced by the NH4+ method caused a rise in aiNa whose maximum amplitude was similar in both groups of muscles. However, the half-time to maximum amplitude was increased by about 56% in diabetic muscles. These results are consistent with a diabetes-induced decrease in the activity of the sarcolemmal Na(+)-H+ exchange.

?-Adrenoceptor-mediated depolarization of the resting membrane in guinea-pig papillary muscles: changes in intracellular Na+, K+ and Cl? activities

Effects of beta-adrenergic stimulation on the membrane potential and intracellular Na+, K+ and Cl- activities were examined in isolated guinea-pig ventricular muscles using conventional and ion-selective electrodes. Isoproterenol in concentrations of 30 nM - 1 microM produced a transient depolarization followed by a slight hyperpolarization in electrically stimulated or quiescent papillary muscles. The negative logarithm of the concentration producing 50% maximum effect (pD2) for the membrane-depolarizing effect of isoproterenol was smaller than that for the positive inotropic effect, suggesting that a higher level of cAMP accumulation is required to produce the transient depolarization. Whereas the isoproterenol(1 microM)-induced depolarization was not blocked by tetrodotoxin (10 microM), nifedipine (10 microM), Cs+ (5 mM), Ba2+ (0.3 mM), amiloride (1 mM) or ouabain (10 microM), it was significantly attenuated by anthracene-9-carboxylic acid (1 mM), a Cl(-)-channel blocker. Intracellular K+ activity increased, whereas intracellular Na+ activity slightly decreased during the transient depolarization. Intracellular Cl- activity significantly decreased during the isoproterenol-induced depolarization of the resting membrane. These results suggest that an inward current resulting from outward Cl- movement, rather than inward Na+ movement, may be involved in the beta-adrenoceptor-mediated membrane depolarization.

Muscarinic receptor stimulation and cyclic AMP‐dependent effects in guinea‐pig ventricular myocardium

1. The effect of carbachol on force of contraction, contraction duration, intracellular Na+ activity and cyclic AMP content was studied in papillary muscles of the guinea-pig exposed to isoprenaline or the phosphodiesterase inhibitor 3-isobutyl, 1-methyl xanthine (IBMX). The preparations were obtained from reserpine-pretreated animals and were electrically driven at a frequency of 0.2 Hz. 2. Isoprenaline (10 nM) and IBMX (100 microM) produced comparable positive inotropic effects of 9.8 and 9.7 mN, respectively. Carbachol (3 microM) attenuated the inotropic effects by 82% (isoprenaline) and by 79% (IBMX). The shortening of contraction duration which accompanied the positive inotropic effect of isoprenaline (by 14.9%) and of IBMX (by 22.4%) was not significantly affected by 3 microM carbachol. 3. The positive inotropic effect of 10 nM isoprenaline and of 100 microM IBMX was accompanied by an increase in cellular cyclic AMP content of 58 and 114%, respectively. Carbachol (3 microM) failed to reduce significantly the elevated cyclic AMP content of muscles exposed to either isoprenaline or IBMX. 4. In the quiescent papillary muscle, isoprenaline (10 nM) and IBMX (100 microM) reduced the intracellular Na+ activity by 28 and 17%, respectively. This decline was not influenced by the additional application of 3 microM carbachol. 5. The results demonstrate that muscarinic antagonism in guinea-pig ventricular myocardium exposed to cyclic AMP-elevating drugs is restricted to force of contraction. The underlying mechanism does not apparently involve the cytosolic signal molecule cyclic AMP.

Muscarinic receptors mediate negative and positive inotropic effects in mammalian ventricular myocardium: differentiation by agonists

The concentration-dependence of the negative and positive inotropic effect of choline esters and of oxotremorine was studied in isometrically contracting papillary muscles of the guinea-pig. The preparations were obtained from reserpine-pretreated animals and were electrically driven at a frequency of 0.2 Hz. In the presence of the phosphodiesterase inhibitor 3-isobutyl-1-methyl xanthine (IBMX, 100 mumol l-1), choline esters and oxotremorine produced concentration-dependent negative inotropic effects. Oxotremorine exhibited the highest negative inotropic potency (with a half-maximal effective concentration, EC50, of 20 nmol l-1) followed by carbachol (139 nmol l-1), methacholine (490 nmol l-1), acetylcholine in the presence of 10 mumol l-1 physostigmine (1.36 mumol l-1) and bethanechol (10 mumol l-1). Atropine was a competitive antagonist of the negative inotropic effects. Carbachol and oxotremorine decreased Vmax, overshoot and duration of slow Ca2+-dependent action potentials which had been elicited in the presence of 100 mumol l-1 IBMX. Choline esters produced a concentration-dependent positive inotropic effect. With an EC50 of 32 mumol l-1, carbachol was the most potent compound, followed by methacholine (35 mumol l-1), acetylcholine in the presence of 10 mumol l-1 physostigmine (46 mumol l-1) and bethanechol (142 mumol l-1). Compared to carbachol and methacholine which increased force by 100% of control, the increase induced by acetylcholine and bethanechol was only 64 and 58%, respectively. Atropine shifted the concentration-effect curves of all choline esters to higher concentrations. Choline esters caused intracellular Na+ activity to increase in the quiescent papillary muscle. This effect was reversed by atropine. Oxotremorine produced a small concentration-dependent positive inotropic effect (about 30% of the maximal effect of carbachol) which was resistant to atropine. Oxotremorine was a potent inhibitor of the positive inotropic effect of choline esters, and did not cause an increase in intracellular Na+ activity in the quiescent papillary muscle. The results show that muscarinic receptors of the ventricular myocardium mediate two inotropic effects, which are opposite in direction and differ in their concentration-dependence by a factor of 100. Although agonists differentiate between both inotropic effects, it is unknown whether the receptors involved represent receptor states or separate receptor subpopulations. The negative inotropic effect of choline esters and of oxotremorine can be best explained by adenylate cyclase inhibition. While stimulation of phosphoinositide hydrolysis might have been responsible for the positive inotropic effect of choline esters via modulation of cation-fluxes across the cell membrane, such a mechanism was not involved in the positive inotropic effect of oxotremorine.

Intracellular chloride activity in rabbit papillary muscle: effect of serum.

The intracellular chloride activity (aiCl), measured with Cl-selective microelectrodes on stimulated rabbit papillary muscles (1 Hz) incubated in serum, was 7.2 +/- 2.2 mM (48 measurements). Under the same condition, on the quiescent muscle, aiCl was 7.5 +/- 2.8 mM (45 measurements). The membrane potential of quiescent papillary muscles and diastolic potential of stimulated papillary muscles were -79.0 +/- 0.7 (50 measurements) and -83.5 +/- 0.5 mV (50 measurements), respectively. The experimental conditions were chosen to reproduce the in vivo conditions where the Cl equilibrium potential is close to the membrane potential or to the diastolic potential. After correcting for cytoplasmic interference (4 mM) on the aiCl measurements, the Cl equilibrium potential (ECl) was -84 mV. In conclusion, the Cl distribution in cardiac cells bathed in serum is passive as for in vivo cardiac cells.

Stretch-induced increase in resting metabolism of isolated papillary muscle

A mathematical model of oxygen diffusion into quiescent papillary muscles in vitro is developed. The model incorporates a continuous sigmoidal function relating the rate of oxygen consumption and the partial pressure of oxygen within the tissue. The behavior of the model is explored over a wide range of external oxygen partial pressures, oxygen consumption/partial pressure relations, oxygen diffusivities, muscle dimensions, and resting metabolic rates, while the muscle is subjected to stimulated stretches of various extents in order to test the assertion that the stretch-induced increase in basal metabolic rate observed experimentally implies the existence of an anoxic core region of papillary muscles in vitro. The model predicts the existence of an oxygen diffusion-mediated stretch response of resting papillary muscle metabolism, but one which is quantitatively insignificant compared with experimentally observed values. The classic Hill diffusion model which explicitly predicts an anoxic core, likewise predicts stretch effects of magnitudes smaller than those frequently observed. It is concluded that the increment in basal metabolism of papillary muscles subjected to stretch in vivo cannot be taken as evidence of oxygen diffusion limitation in unstretched preparations.

Non-passive chloride distribution in mammalian heart muscle: micro-electrode measurement of the intracellular chloride activity.

1. Liquid ion-exchanger Cl- -sensitive micro-electrodes were used to make continuous measurements of the intracellular Cl activity, aCli, of quiscent sheep cardiac Purkinje fibres in vitro. 2. aCli was higher than that expected from a passive distribution, (which would have been about 5 mM). It was 3--4 times hiable; EC1 was about 35 mV positive to Em. It was over twice as high in the nominal absence of bicarbonate/CO2 (when the buffer-system was HEPES/O2) but was not always so stable, and ECl was about 20 mV positive to Em. 3. Experiments designed to assess the maximum possible error likely to occur in the measurement of aCli showed that this could not be large and that the estimates of ECl were accurate to within 8 mV. 4. The ability of Cl to move down both concentration and potential gradients was established by demonstrating a loss of aCli in Cl-free solutions and a gain when Em was depolarized positive to ECl in high-K solutions. In both cases, the changes were complete within about 100--160 min. 5. The decline of aCli in Cl-free solutions (glucuronate-substituted) was not significantly affected by changes of [Ca]o from 0 to 12 mM or by the depolarizations of Em of up to 60 mV that sometimes occurred in low or zero [Ca]o. 6. Only 2--3 mM-aClo was sufficient to impede substantially the ready loss of aCli in HEPES-buffered solutions. 7. In high-K solutions (45 mM), Cl appeared to be passively distributed since, at equilibrium, Em and ECl differed by less than 2 mV. 8. In HEPES-buffered Tyrode, ECl of quiescent papillary muscle of the guinea-pig was, on average, 39 mV positive to Em. 9. It is concluded that liquid ion-exchanger Cl- -sensitive micro-electrodes are suitable for studying the Cl regulation of sheep Prukinje fibres, and probably of other cardiac tissues. The measurements of resting aCli are quite accurate when using either HEPES or bicarbonate-buffered Tyrode. The results are discussed in relation to estimates of the apparent membrane Cl permeability under various conditions and the possible existence of an inwardly directed 'Cl pump'.

Cat heart muscle in vitro. II. The steady state restpotential in quiescent papillary muscles.

The steady state transmembrane resting potential difference (V(m)) has been measured in quiescent papillary muscles. V(m) was determined as a function of the external K concentration in Cl and SO(4) solutions and compared with the K equilibrium potential. Other measurements were made after replacement of external Na by choline, K by Rb and Cs, and Cl by SO(4), CH(3)SO(4), and NO(3). Effects on V(m) of albumin, temperature, and variation in internal K concentration are described.

The rate of oxygen uptake of quiescent cardiac muscle.

The rate of oxygen uptake of quiescent papillary muscle of the cat heart has been determined in a flow respirometer with the use of the oxygen electrode. The apparent rate of oxygen uptake as a function of the diameter of the muscle was also determined. It was found that papillary muscles from cat hearts use oxygen at a rate of 2.84 (microliters/mg. wet weight)/hour at a temperature of 35 degrees C. Such muscles can be adequately supplied by diffusion when their surface is uniformly exposed to an atmosphere containing 95 per cent oxygen only if their diameter is 0.64 mm. or less. Papillary muscles from kitten hearts use oxygen at a rate of 4.05 (microliters/mg. wet weight)/hour at a temperature of 35 degrees C. Such muscles can be adequately supplied by diffusion when their surface is uniformly exposed to an atmosphere containing 95 per cent oxygen only if their diameter is 0.53 mm. or less. If the muscles are small enough to be adequately supplied with oxygen by diffusion, the rate of oxygen uptake does not increase when the muscle is stretched.