The early development of the atrioventricular node and bundle of His in the embryonic chick heart. An electrophysiological and morphological study

The development of the atrioventricular node and bundle of His of embryonic chick hearts was studied by electrophysiological and morphological techniques. The dorsal wall of the AV canal and the interatrial septum were explored to determine if they contribute to the formation of the AV node and bundle of His. The resting membrane and action potentials of the interatrial septum cells were systematically analyzed and found to undergo progressive differentiation with development. The earliest identification of the AV node and upper bundle of His group of cells was achieved at 5 1/2-6 days of development by the electrical recording of their corresponding characteristic action potentials, from a circumscribed area located in the lowest and dorsal segment of the interatrial septum. The morphological and anatomical characterization of the cells was made following electrical recording and labelling with charcoal particles. The earlier AV node and bundle of His responses had similar characteristics to those of the adult heart. It is concluded that the AV node and upper bundle of His cells derive from the low interatrial septum. The possibility that AV canal cells contribute to this event was discarded. The functional relationship of the Av node and bundle of His with other cardiac tissues during the early development of the heart is discussed.     

Intraoperative control of the state of the arrested heart

Monitoring the function of the operated on heart over time based on the principle of recording the effort of the myocardium at rest was tried under the clinical conditions after preliminary experiments on the hearts of rats and dogs. The study included 11 patients operated on for aortal, mitral and multi-valvular prosthetics under pharmacological cardioplegia. Such monitoring enables an objective appraisal of the heart status, Besides, it makes it possible to decide on whether repeated administration of cardioplegic solutions is required, to specify the times of reoxygenation and to prognose the pattern of the heart function recovery after operation is over. The data obtained helped detect some disadvantages of the generally accepted method of pharmacological cardioplegia.     

Spread of excitation through the heart upon electrical stimulation of the atrioventricular valves]

Experiments were conducted on the isolated hearts of rabbits and dogs. Synchronous recording of the electrical activity of the right auricle, the right ventricle, and tricuspid valve demonstrated the existence of definite functional associations between the auricular, ventricular and valvular depolarization. The spread of excitation in the heart in electrical stimulation of the atrio-ventricular valves was investigated. The impulses from the heterotopic excitation focus localized in these valves could be conducted to the other parts of the heart and thus lead to cardiac arrhythmias.     

Dilatory and inotropic effects of corticotropin-releasing factor (CRF) on the isolated heart. Effects on atrial natriuretic peptide (ANP) release.

The effects of CRF administration on cardiac performance, coronary flow and ANP release were investigated in the rat heart. Isolated hearts were perfused at a constant filling pressure according to working heart model with a Krebs-Henseleit solution containing glucose and insulin, saturated with a gas mixture containing 95% O2 and 5% CO2. Administration of CRF via a cannula into the left atrium elicited a prolonged increase in the coronary flow rate and a transient increase in the aortic pressure resulting in an overall increase in the pressure-volume work. The oxygen consumption, after the administration of CRF, increased in accordance with the cardiac effort. No changes were observed in the spontaneous heart rate. Furthermore, administration of CRF induced a short-term increase of ANP release into the coronary perfusate. Our experiments suggest that administration of CRF produces a prolonged dilatory effect on the coronary arteries while producing a transient positive inotropic effect and a transient increase of ANP release on the isolated rat heart.     

Establishment and characterization of a mouse embryonic heart slice preparation.

BACKGROUND: In contrast to isolated cells, the anatomic and functional integrity of tissue slices remains preserved. Aim of the study was to establish the slice technique in embryonic mouse hearts in order to perform physiological and pharmacological investigations of wild-type mice and genetically engineered mouse models of heart disease. METHODS: Ventricular slices (thickness: 300 mum) were cut from agar-embedded embryonic mouse hearts (ED 16.5-18.5) with a vibratome. Histology, immunostaining with markers for apoptosis induction, intracellular recordings with sharp electrodes and field potential recordings using microelectrode arrays were performed to assess viability. RESULTS: Slices exhibited normal histology without prominent signs of apoptosis for at least 24 hours. Intracellular recordings revealed the typical electrophysiological fingerprint of ventricular cardiomyocytes. Field potential recordings proved that adrenergic and muscarinic signaling was preserved. CONCLUSION: Functionally intact heart slices can be generated from murine embryos.     

降钙素基因相关肽在豚鼠冠脉血流量和心脏传导系统作用的比较

本文目的在于深入研究降钙素基因相关肽（ＣＧＲＰ）对豚鼠冠状血流量以及心脏传导系统各部分的作用。采用Ｌａｎｇｅｎｄｏｒｆｆ法灌流心脏,同步记录心脏表面电图和希氏束电活动。观察应用ＣＧＲＰ前后的冠脉流量、自主心率、在相同心房周期下的房室结（ＡＨ）及希浦系传导时间（ＨＶ）、心脏出现３：２文氏传导及２：１房室传导阻滞所需的最长起搏周期（ＰＣＬ３：２,ＰＣＬ２：１）。ＣＧＲＰ（３－３０ｎｍｏｌ／Ｌ）可显著增     

Dynamics of the rythm of the isolated and intact frog heart

This study was designed to examine the changes of the isolated frog's heart rate as a function of the time in the two time intervals: 20-95 min and 95-170 min after hearts were prepared. The heart rate decreased in these intervals quite linear but average slope between 20-75 min was significantly steeper than in the time interval 95-170 min. It was shown that the difference in the heart rate response induced by the increasing temperature in intact animals and isolated hearts partly might be explored by this decrease in the isolated frog's heart rate with time.     

Ventricular fibrillation in hibernators and nonhibernators

Previous studies have shown that there are differences between hibernators and nonhibernators in the susceptibility to ventricular fibrillation. In an attempt to clarify these differences ventricular fibrillation was induced in isolated hearts of the hibernator, the woodchuck, Marmota monax by cooling, warming, puncture, and by norepinephrine administration. It was shown that the hearts of the winter animals were completely resistant toward the ventricular fibrillation inducing agents, which was not the case for the hearts from summer, active animals. Further, the hearts of another hibernator, the hedgehog, Erinaceus europaeus, and guinea pig, Cavia porcellus, were studied electrophysiologically in anesthetized animals with open chests and with bipolar electrodes attached to the epicardium. During pacing it was shown that the hedgehog had a higher stimulus threshold and a lower maximal following frequency than the guinea pig. The summer hedgehogs showed resistance toward both ventricular premature beats and ventricular fibrillation. Sixty percent of the summer hedgehogs and 100 percent of the winter hedgehogs and guinea pigs developed ventricular fibrillation. The threshold for ventricular fibrillation was highest for summer hedgehogs. The effective refractory period of papillary muscle of summer hedgehogs was shorter than that of guinea pigs. The force frequency relationship of the isolated papillary muscle showed a greater degree of independence in the hedgehog than in the guinea pig. Consequently, the results show that the heart of the hibernator is more arrhythmia resistant than the heart of the nonhibernator, although there are seasonal differences.     

The ionic basis of electrical activity in embryonic cardiac muscle

The intracellular sodium concentration reported for young, embryonic chick hearts is extremely high and decreases progressively throughout the embryonic period, reaching a value of 43 mM immediately before hatching. This observation suggested that the ionic basis for excitation in embryonic chick heart may differ from that responsible for electrical activity of the adult organ. This hypothesis was tested by recording transmembrane resting and action potentials on hearts isolated from 6-day and 19-day chick embryos and varying the extracellular sodium and potassium concentrations. The results show that for both young and old embryonic cardiac cells the resting potential depends primarily on the extracellular potassium concentration and the amplitude and rate of rise of the action potential depend primarily on the extracellular sodium concentration.     

Dual probe fluorescence monitoring of intracellular free calcium during ischemia in mouse heart by using continuous compensation for pH dependence of the dissociation constant of Fura-2, and the interference of myoglobin

Mitochondrial damage is the main source of cellular injury upon ischemia-reperfusion, and calcium loading has been implicated in this phenomenon. The use of optical probes for calcium monitoring of the intact heart is hampered by internal filter effects of intracellular hemoproteins, endogenous fluorescence, and their sensitivity to pH. We describe here a method for measurement of intracellular free calcium in isolated myoglobin-deficient perfused mouse hearts under conditions of large intracellular pH fluctuations by simultaneous fluorescence monitoring of the calcium-probe Fura-2 and the pH probe BCECF through dual wavelength excitation of both probes. In myoglobin-containing mouse heart endogenous chromophores interfere with Fura-2 fluorometry. It is shown that a paradoxical decrease in Fura-2 fluorescence occurs during ischemia in isolated mouse hearts. Simultaneous recording of BCECF fluorescence (calibrated against pH measurement with phosphorus NMR) and data reduction based on continual recalculation of the apparent dissociation constant of the calcium-probe complex revealed that a marked increase in intracellular free calcium occurs, and that the Fura-2 fluorescence decrease was caused by an increase in dissociation constant due to intracellular acidification. Intracellular free calcium rose almost linearly during a 20-min period of ischemia and returned to basal values rapidly upon the commencement of perfusion.     

Chronic alcoholic cardiomyopathy. Protection of the isolated ischaemic working heart by ribose

The effects of ribose on the pre- and post-ischaemic functional performance of the isolated working heart from 24 month old chronically alcoholic animals was investigated. The improved perfusion model permitted the isolated heart to perform work analogous to that of the normal physiological load, in a system where systemic pressure and atrial pressure could be altered over a wide range and oxygen loss from the perfusion fluid was a minimum. There was a remarkable improvement in the performance of isolated hearts taken from alcoholic animals that were perfused with 1.7 mM ribose both before and after a 25.0 min period of global myocardial ischaemia (at 25 degrees C), however ribose treatment did not greatly affect the performance of hearts of isocaloric control aged rats. Chronic alcohol consumption significantly affected heart performance, causing a marked reduction in both cardiac and work output. After ischaemia the work of all hearts was notably decreased; there was no work output in untreated hearts of alcoholic animals, whereas in hearts of alcoholic animals treated with ribose work output was only decreased by 35%. The acute response to ribose by hearts of aged chronically alcoholic animals suggests a role for this compound as a positive inotropic agent and clearly indicates the beneficial potential of ribose for inclusion in cardioplegic solutions or for infusion in alcoholic subjects showing signs of heart failure or chronic heart disease.     

Dual probe fluorescence monitoring of intracellular free calcium during ischemia in mouse heart by using continuous compensation for pH dependence of the dissociation constant of Fura-2, and the interference of myoglobin

Mitochondrial damage is the main source of cellular injury upon ischemia–reperfusion, and calcium loading has been implicated in this phenomenon. The use of optical probes for calcium monitoring of the intact heart is hampered by internal filter effects of intracellular hemoproteins, endogenous fluorescence, and their sensitivity to pH.We describe here a method for measurement of intracellular free calcium in isolated myoglobin-deficient perfused mouse hearts under conditions of large intracellular pH fluctuations by simultaneous fluorescence monitoring of the calcium-probe Fura-2 and the pH probe BCECF through dual wavelength excitation of both probes. In myoglobin-containing mouse heart endogenous chromophores interfere with Fura-2 fluorometry.It is shown that a paradoxical decrease in Fura-2 fluorescence occurs during ischemia in isolated mouse hearts. Simultaneous recording of BCECF fluorescence (calibrated against pH measurement with phosphorus NMR) and data reduction based on continual recalculation of the apparent dissociation constant of the calcium-probe complex revealed that a marked increase in intracellular free calcium occurs, and that the Fura-2 fluorescence decrease was caused by an increase in dissociation constant due to intracellular acidification. Intracellular free calcium rose almost linearly during a 20-min period of ischemia and returned to basal values rapidly upon the commencement of perfusion.     

New high-frequency catheter technique for His bundle ablation in dogs

A new method is presented for the production of complete atrioventricular heart block. It consists of a special catheter, which is inserted into the right atrium via a femoral vein and positioned in the region of the His bundle for His bundle potential recording. Production of heart block is achieved by a high frequency current pulse from an electrocautery unit.     

Role of oxidative stress in alterations of mitochondrial function in ischemic-reperfused hearts

To study the mechanisms of mitochondrial dysfunction due to ischemia-reperfusion (I/R) injury, rat hearts were subjected to 20 or 30 min of global ischemia followed by 30 min of reperfusion. After recording both left ventricular developed pressure (LVDP) and end-diastolic pressure (LVEDP) to monitor the status of cardiac performance, mitochondria from these hearts were isolated to determine respiratory and oxidative phosphorylation activities. Although hearts subjected to 20 min of ischemia failed to generate LVDP and showed a marked increase in LVEDP, no changes in mitochondrial respiration and phosphorylation were observed. Reperfusion of 20-min ischemic hearts depressed mitochondrial function significantly but recovered LVDP completely and lowered the elevated LVEDP. On the other hand, depressed LVDP and elevated LVEDP in 30-min ischemic hearts were associated with depressions in both mitochondrial respiration and oxidative phosphorylation. Reperfusion of 30-min ischemic hearts elevated LVEDP, attenuated LVDP, and decreased mitochondrial state 3 and uncoupled respiration, respiratory control index, ADP-to-O ratio, as well as oxidative phosphorylation rate. Alterations of cardiac performance and mitochondrial function in I/R hearts were attenuated or prevented by pretreatment with oxyradical scavenging mixture (superoxide dismutase and catalase) or antioxidants [N-acetyl-L-cysteine or N-(2-mercaptopropionyl)-glycine]. Furthermore, alterations in cardiac performance and mitochondrial function due to I/R were simulated by an oxyradical-generating system (xanthine plus xanthine oxidase) and an oxidant (H(2)O(2)) either upon perfusing the heart or upon incubation with mitochondria. These results support the view that oxidative stress plays an important role in inducing changes in cardiac performance and mitochondrial function due to I/R.     

Differences in the reaction of rat and guinea pig hearts to ischemia and reperfusion

The response of rat and guinea-pig hearts to ischemia and reperfusion has been studied in identical conditions. Total 15-min ischemia of isolated rat hearts at 36 degrees C induced an almost 3-fold rise in isovolumic left ventricular diastolic pressure as well as a fall in the developed pressure and heart rate. Guinea-pig hearts, in the same conditions, exhibited a more steep fall in heart rate, with no rise in diastolic pressure. With constant heart rate produced by electrical stimulation at 4 Hz, the difference between two groups remained unchanged, while a more rapid fall in developed pressure in guinea-pig hearts coincided with a more profound fall in extracellular pH and almost a 2-fold rise in extracellular K+ activity. Rapid elimination of K+ and H+ at the early stages of reperfusion was followed by fibrillation in the majority of guinea-pig hearts, while no fibrillation was observed in rat hearts.     

A new floating sensor array to detect electric near fields of beating heart preparations

A new flexible sensor for in vitro experiments was developed to measure the surface potential, Φ, and its gradient, E (electric near field), at given sites of the heart. During depolarisation, E describes a vector loop from which direction and magnitude of local conduction velocity θ can be computed. Four recording silver electrodes (14 μm × 14 μm) separated by 50 μm, conducting leads, and solderable pads were patterned on a 50 μm thick polyimide film. The conductive structures, except the electrodes, were isolated with polyimide, and electrodes were chlorided. Spacer pillars mounted on the tip fulfil two functions: they keep the electrodes 70 μm from the tissue allowing non-contact recording of Φ and prevent lateral slipping. The low mass (9.1 mg) and flexibility (6.33 N/m) of the sensor let it easily follow the movement of the beating heart without notable displacement. We examined the electrodes on criteria like rms-noise of Φ, signal-to-noise ratio of Φ and E, maximum peak-slope recording dΦ/dt, and deviation of local activation time (LAT) from a common signal and obtained values of 24–28 μV, 46 and 41 dB, 497–561 V/s and no differences, respectively. With appropriate data acquisition (sampling rate 100 kHz, 24-bit), we were able to record Φ and to monitor E and θ on-line from beat-to-beat even at heart rates of 600 beats/min. Moreover, this technique can discriminate between uncoupled cardiac activations (as occur in fibrotic tissue) separated by less than 1 mm and 1 ms.     

Response to beta-blockers in maternal and fetal rat hearts in vitro.

The effects of beta-blockers on maternal and fetal heart rates have been assessed by comparing isoprenaline concentration-heart rate relationships of hearts isolated from pregnant rats. The normal and maximal heart rates obtained for the maternal and fetal hearts were similar to published data. A slightly but significantly higher concentration of isoprenaline was required to produce 50% of the maximal response of fetal hearts than maternal hearts, suggesting that fetal hearts were less sensitive to isoprenaline than the maternal hearts. The beta-blockers used (propranolol, labetalol, metoprolol and atenolol) all showed a lower affinity to the beta-receptors of fetal hearts than those of maternal hearts, as indicated by significant differences in the pA2 values. Given the similar effects of the beta-blockers in the maternal and fetal hearts it is concluded that pharmacokinetic considerations and beta-blocker selectivity should be used as the basis of choice when treating maternal hypertension during pregnancy.     

Evidence for new forms of cardiac myosin heavy chains in mechanical heart overloading and in ageing

Heavy chains of myosin rods and subfragment 1 were isolated from normal hearts and from mechanically overloaded hearts of young and older rats. These myosin heavy-chain fragments were cleaved by cyanogen bromide or partially proteolysed by pronase and by chymotrypsin after denaturation with sodium dodecyl sulfate. The peptides, analyzed by electrophoresis on a one-dimensional polyacrylamide slab gel, varied depending on the origin of the cardiac myosin heavy chains. Some bands present in the peptide patterns of the normal heart of young rats were missing from the pattern of greatly hypertrophied hearts and vice versa. We conclude that mechanical overloading of the heart stimulates the synthesis of cardiac myosin 'isozyme' with a heavy-chain primary structure which is different from that observed in the normal heart of young rat. The patterns from myosin heavy-chain peptides from the hearts of older rats were different from those for peptides from young rat hearts; these results also indicate the presence of a new myosin heavy chain specific to ageing. No difference was detected between the peptide patterns of heavy chains isolated from hypertrophied hearts of young and older rats, and those isolated from normal hearts of older rats.     

Electrophysiological effects of the aqueous extract of Averrhoa carambola L. leaves on the guinea pig heart

This work aims to describe some electrophysiological changes promoted by the aqueous extract (AEx) from Averrhoa carambola leaves in guinea pig heart. The experiments were carried out on isolated heart or on right atrium-ventricle preparations. In 6 hearts, the extract induced many kinds of atrioventricular blocks (1st, 2nd, and 3rd degrees); increased the QT interval from 229+/-23 to 264+/-19 ms; increased the QRS complex duration from 27+/-3.1 to 59+/-11 ms, and depressed the cardiac rate from 136+/-17 to 89+/-14b pm. Furthermore, it decreased the conduction velocity of atrial impulse (17+/-3%); reduced the intraventricular pressure (86+/-6%), and increased the conduction time between the right atrium and the His bundle (27+/-6.5%). The conduction time from the His bundle to the right ventricle was not altered. Atropine sulfate did not change either the electrocardiographic parameters or the intraventricular pressure effects promoted by the A. carambola AEx. Based on these results, the popular use of such extracts should be avoided because it can promote electrical and mechanical changes in the normal heart.     

Effect of brief vascular perfusion on the ultrastructure and activity of mitochondria isolated from the rat heart

Summary Mitochondria isolated from heart tissue after a 1-min perfusion with Hanks medium were found to have significantly lower rates of State-3 respiration and respiratory control ratios compared to mitochondria isolated from non-perfused hearts. Examination of the mitochondrial preparations by electron microscopy revealed that a large proportion of the mitochondria isolated from perfused heart tissue were swollen and broken compared to mitochondria from non-perfused hearts.