Dose-dependent reduction of cardiac transmembrane potential by high-intensity electrical shocks

Cardiac tissue dysfunction can result from high-intensity electrical shocks and is manifested as changes in transmembrane potential ( V m). Ten-millisecond shock pulses (SPs) of varying intensity and polarity were applied to frog ventricle in diastole, and V m was quantified directly under the stimulating electrode by an optical method using voltage-sensitive dye. As SP intensities were increased, the shock-induced action potential (AP) plateau and AP amplitude (APAs) decreased sigmoidally toward 75–85% of the control AP amplitude (APAc) and zero, respectively. APAswas shifted toward lower current densities for anodal compared with cathodal SPs (half-maximal values 185 and 238 mA/cm2, respectively; P = 0.02). Recovery of APAs was marginally significant 1 s after SP delivery ( P = 0.063). The peak change in V mduring SP (across all intensity levels) was −200% APAc for anodal and +125% APAc for cathodal pulses. In conclusion, we show that SP reduces APA in a sigmoidal fashion at strengths >10–20 × diastolic threshold and is more deleterious for anodal polarities.

Cyclic nucleotide phosphodiesterases from frog atrial fibers: isolation and drug sensitivities

The cyclic nucleotide phosphodiesterase (PDE) forms present in frog atrial fibers were isolated and characterized by their drug sensitivities. DEAE-sephacel chromatography of cytosolic PDE activity resolved three major PDE forms: peak A hydrolyzed both adenosine 3',5'-cyclic monophosphate (cAMP) and guanosine 3',5'-cyclic monophosphate (cGMP) and was activated by calcium-calmodulin (PDE I); peak B also hydrolyzed both cAMP and cGMP but was activated by 5 microM cGMP (PDE II); peak C specifically hydrolyzed cAMP (PDE IV). Rolipram specifically inhibited PDE IV (Ki = 1.1 microM), whereas dipyridamole potently inhibited both PDE II (Ki = 4.6 microM) and PDE IV (Ki = 0.8 microM). Atrial fiber PDE I was preferentially inhibited by zaprinast (Ki = 10 microM). 3-Isobutyl-1-methyl xanthine (IBMX) and theophylline inhibited nonspecifically all three different enzymes. The positive inotropic drug CI 930 only inhibited the different isolated atrial PDE forms at concentrations greater than 200 microM. However, under assay conditions for which PDE IV was specifically inhibited (presence of 100 microM rolipram), an IC50 of 17 microM for CI 930 was observed on the remaining 26% cAMP hydrolytic activity of peak C (which could represent a cGMP-inhibited PDE form: PDE III). The same PDE forms were also found in frog ventricle. The major difference between frog atrial fiber (and ventricular tissue) PDEs and mammalian cardiac PDEs is that the main cytosolic cAMP-specific hydrolytic activity in frog heart is due to PDE IV rather than PDE III. Rolipram, dipyridamole, and zaprinast might be useful tools to investigate the participation of cAMP in frog atrial contraction (unpublished observations).

Use of frog ventricle to examine mechanical and electrical activity of heart.

This report describes a Langendorff heart preparation, which utilizes frog rather than mammalian hearts to demonstrate both mechanical and electrical events of the cardiac cycle. The preparation is durable in that it can be maintained for several hours while being perfused with room-temperature solutions that are not supplemented with oxygen. Ventricular perfusion is achieved via a fluid-filled reservoir coupled to a truncus arteriosus catheter advanced into the ventricle. By varying the height of the reservoir relative to the heart, changes in ventricular pressure and the rate of ventricular pressure change during systole can be recorded by way of a side port on the catheter tubing. The former is indicative of Starling's law of the heart, whereas the latter is reflective of the contractility. Electrical activity of the heart can be observed by measuring injury potentials via a needle electrode inserted into the ventricle. These are extracellular potentials that reflect ventricular action potentials. In addition, surface electrodes placed on the ventricle can be used to detect an integrated ventricular electrocardiogram. Recording of ventricular pressure simultaneously with at least one of these two electrical recordings allows a direct comparison of electrical and mechanical events of the heart. In summary, the Langendorff frog heart preparation is economical in terms of both financial cost and simplicity; yet it enables a thorough examination of both electrical and mechanical properties of the heart either as a student lab exercise or as a classroom demonstration.

Frog ventricle: participation of SR in excitation-contraction coupling

Activation of the first beat (B1) following a 60-s pause is diminished in isometrically contracting frog ventricular strips, in contrast to the augmentation documented for sarcoplasmic reticulum (SR)-dependent mammalian myocardium. However, treatment of frog ventricular strips with ouabain, an indirect inhibitor of the sarcolemmal Na+-Ca2+ exchanger, selectively enhanced postpause beats suggesting that in the absence of ouabain significant extrusion of cellular Ca2+ occurred during the pause. Because resting tension did not increase during the pause in ouabain-treated strips, the nonextruded Ca2+ must have been sequestered into a compartment such as SR. Steady-state beats were not affected by ouabain; its actions appeared to be separate from its known positive inotropism. Caffeine, a direct SR stimulus, initially enhanced B1 and subsequently decreased activation of all beats, which was consistent with initial augmentation of SR Ca2+ release and subsequent depletion of SR Ca2+ stores. Ouabain both potentiated the stimulatory effects and blocked the inhibitory effects of caffeine, suggesting that ouabain increased Ca2+ stores in the same intracellular Ca2+ pool as that acted on by caffeine, the SR. Ryanodine, an inhibitor of SR in mammalian myocardium, did not affect activation of frog myocardium. SR may be an important site for activator Ca2+ cycling in frog myocardium under control conditions as well as after long diastolic intervals in the presence of ouabain.