Differentiation of the atrioventricular node, the atrioventricular bundle and the bundle branches in the bovine heart: an immunohistochemical and enzyme histochemical study

The previous observations of differences between different cardiac regions (ventricular myocardium, atrial myocardium, Purkinje fibre system) with respect to the maturation of the M-line region and the establishment of mature metabolic characteristics, have been extended. It was found that M-line maturation proceeds differently also between different regions of the conduction system. The M-line proteins, myomesin and MM-creatine kinase, were detected earlier, by means of immunohistochemistry, in the AV bundle and bundle branch cells than in the AV node cells. Also, a difference was observed in large foetuses. Striations in the AV node were less evident than in the AV bundle and the bundle branches in sections incubated with antibodies against myomesin as well as against MM-creatine kinase. Using enzyme histochemistry it was observed that the differences in metabolic properties between the AV node, the AV bundle and the bundle branches on the one hand, and the ordinary myocardium on the other, of adult hearts, are not established at the early stages. No clear difference in activity of succinate dehydrogenase was seen between the conduction tissues and the ordinary myocardium in the foetal hearts, while the conduction tissues showed a lower activity in the adult hearts. Furthermore, the pattern of activity of mitochondrial glycerol-3-phosphate dehydrogenase between the conduction tissues and the atrial and ventricular myocardium was quite different in early foetal stages compared with the adult stage.     

The atrioventricular node and bundle in the ferret heart: a light and quantitative electron microscopic study.

The cells of the atrioventricular (AV) junction in the ferret heart were examined using light microscopy, a wax-model reconstruction and quantitative electron microscopy to determine their organization and characteristics. A series of subdivisions of the specialized tissues of the AV junction was apparent at both the light and electron microscopic levels. A transitional zone was observed interposed between the atrial muscle cells and the AV node. The AV node consisted of a coronary sinus portion, a superficial portion and a deep portion. The AV bundle had a segment above the anulus fibrosus, a segment which penetrated the right fibrous trigone, a non-branching segment below the anulus fibrosus and a branched segment. At the ultrastructural level the AV junctional conduction tissues had fewer irregularly oriented myofibrils than did working atrial myocardial cells. T-tubules, present in atrial muscle cells, were not observed in the modified muscle cells of the AV node and bundle. Conventional intercalated discs also were not observed between the cells of the AV node or the AV bundle. Atrial myocardial cells had the highest percentage of the plasma membrane occupied by desmosomes, fasciae adherentes and gap junctions. The AV bundle cells had the highest percentage of appositional surface membrane and a relatively large fraction of plasma membrane occupied by gap junctions. Cells of the superficial portion of the AV node had the smallest percentage of the plasma membrane composed of gap junctions, desmosomes or fasciae adherentes, as well as the smallest fraction of the cell membrane apposed to adjacent cells. The stereological data indicate that the most useful distinguishing characteristic between atrial muscle cells and conduction cells was that a smaller percentage of the conduction cell sarcoplasm was occupied by mitochondria and myofibrils. The most useful characteristics that could be used to differentiate between the regions of the AV junctional conduction tissues were the amounts and types of surface membrane specializations in the respective cell types.     

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.     

A morphometric analysis of the structure of the atrioventricular node of the sheep heart]

The work deals with the geometry of the structure of different sections of the atrio-venticular (A-V) node of the sheep's heart and measurement of the diameter of fibres in its three zones: atrio-nodal (A-N), the node proper (N) and transition of the node into His bundle (NH). The revealed difference in the diameter of contacting fibres of N and NH areas under worse conditions of conducting may contribute to block of excitation. The geometrical features of A-N and N areas contribute to prolonged delay of excitation and geometrical parameters of the site of contact between N and NH zones to its block.     

Quantification of concealed conduction in the atrioventricular node

Twenty-eight anaesthetized open-chest mongrel dogs were used. Programmed atrial pacing was used and Hisian electrograms recorded through endocavitary electro-catheters to study and quantify the concealed conduction of non-transmitted atrial impulses in the A-V node. An exponential model was used in three situations to quantify the nodal conduction during incremental atrial pacing: a) during 1:1 conduction, b) during 2:1 nodal block, and c) during pacing, coupling an atrial impulse delivered at fixed intervals and blocked in the A-V node to each transmitted impulse. The relation between intranodal conduction times was analyzed both with and without the presence of blocked impulses, and the quotient between the obtained functions in situations b, c and situation a was determined. In a subgroup of 13 dogs the study was repeated following pharmacological block of the autonomic nervous system. In dogs with autonomic block, this relation always tended to decrease when the atrial pacing rate increased. The variations in the group of dogs with intact autonomic nervous systems were not homogeneous. During pacing with coupled block impulses, the progressive removal of conduction curves obtained for each coupling interval with respect to those obtained during 1:1 transmission, expresses the interval with respect ot those obtained during 1:1 transmission, expresses the lesser influence of the blocked impulses on decreasing their coupling interval.     

The development of innervation in the rat atrioventricular node

Summary The problem of development of the innervation of the rat atrioventricular node has been investigated by electron microscopy. Nerve bundles appear in relation to the node as early as the second postnatal day and vesiculated axons are seen throughout the entire node by the fourth day. Intimate contacts between nodal cells, axons and terminal varicosities are frequently observed.Use of the 5-hydroxydopamine tracer technique has enabled the identification of both cholinergic and adrenergic axons. It is concluded that the node has a dual innervation although cholinergic endings far outnumber those classified as adrenergic on the sixth postnatal day.These results are quite different to earlier findings made at the light microscope level and the discrepancies are discussed with respect to the histochemical techniques used. The suggestion that nodal differentiation is induced by nerves is considered in relation to the differences in cholinesterase activity exhibited by nodal cells during normal development and following neonatal sympathectomy.     

Robotic-assisted cryosurgical treatment of atrioventricular node reentrant tachycardia

Atrioventricular nodal reentrant tachycardias typically arise from the existence of variable refractoriness in fast and slow conduction pathways within the triangle of Koch, which provide input to the atrioventricular node. Standard therapy includes medical management and catheter-based ablation procedures. Robotic-assisted, minimally invasive cryosurgical modification of the atrioventricular node can provide definitive therapy for patients who fail traditional therapy. A 65-year-old man presented with a several-year history of recurrent atrioventricular nodal reentrant tachycardia. Despite medical management and attempted percutaneous ablation, the patient remained symptomatic with weekly episodes. Access was via a 4-cm right anterolateral thoracotomy and peripheral perfusion. The da Vinci S robotic system was used to manipulate the cryoprobe (CryoMaze Probe; ATS Medical, Plymouth, MN USA). A series of spot freezes (tip 60°C) were made along the boundaries of the triangle of Koch until transient complete heart block was achieved and nodal rhythm was recovered. At follow-up 3 weeks postoperatively, the patient was asymptomatic in first-degree heart block. Robotic-assisted cryosurgical atrioventricular node ablation is an effective, minimally invasive treatment for patients with atrioventricular nodal reentrant tachycardia.     

Simulation analysis of conduction of excitation in the atrioventricular node

The excitation conduction in the atrioventricular node was simulated based on the spatially discrete model of the heart proposed in an earlier paper (Kawato et al., 1986). We constructed a model system composed of the atrium, the atrioventricular node and the Purkinje fiber. Coupling coefficients between these tissues were quantitatively estimated from experimental data on size and membrane capacitance of the three kinds of cardiac cells. We found the following three important features in the simulated excitation conduction along the atrioventricular node. First, shape of action potential was found to be different at different locations of the atrioventricular node although the membrane properties were assumed uniform through the atrioventricular node. Our analysis suggests that the difference in the action potential waveforms observed by Paes de Carvalho & De Almedia (1960) can be ascribed to the electrical influences of the atrium and the His bundle on the atrioventricular node. Second, when the excitation wavefront invaded the atrioventricular node from the atrium, a step was observed in the depolarization phase of the action potential at the atrioventricular node neighboring with the atrium. Janse found a similar step in the real experiment (1969). It is revealed that this step is caused by termination of the junctional current which flows from the atrium to the atrioventricular node. Finally, we found that the conduction velocity measured near the boundary between the atrium and the atrioventricular node was lower than that in the middle part of the atrioventricular node, which is in accordance with the experimental observation by Scher et al. (1959).     

Role of pacemaking current in cardiac nodes: insights from a comparative study of sinoatrial node and atrioventricular node

Cardiac pacemaking in the sinoatrial (SA) node and atrioventricular (AV) node is generated by an interplay of many ionic currents, one of which is the funny pacemaker current (I_f). To understand the functional role of I_f in two different pacemakers, comparative studies of spontaneous activity and expression of the HCN channel in mouse SA node and AV node were performed. The intrinsic cycle length (CL) is 179±2.7 ms (n=5) in SA node and 258±18.7 ms (n=5) in AV node. Blocking of I_f current by 1 μmol/L ZD7288 increased the CL to 258±18.7 ms (n=5) and 447±92.4 ms (n=5) in SA node and AV node, respectively. However, the major HCN channel, HCN4 expressed at low level in the AV node compared to the SA node. To clarify the discrepancy between the functional importance of I_f and expression level of HCN4 channel, a SA node cell model was used. Increasing the I_f conductance resulted in decreasing in the CL in the model, which explains the high pacemaking rate and high expression of HCN channel in the SA node. Resistance to the blocking of I_f in the SA node might result from compensating effects from other currents (especially voltage sensitive currents) involved in pacemaking. The computer simulation shows that the difference in the intrinsic CL could explain the difference in response to I_f blocking in these two cardiac nodes.     

Generation of cardiomyocytes by atrioventricular node cells in long-term cultures

Turnover of cardiac pacemaker cells may occur during the lifetime of the body, and we recently raised the hypothesis that specialized cardiac cells have in common the potential to generate cardiomyocytes from fibroblasts. To examine this hypothesis, we analyzed the ability of atrioventricular node cells (AVNCs) to generate functional cardiomyocytes in long-term culture. AVNCs were isolated from adult guinea pig hearts and cultured for up to three weeks. Under phase-contrast microscopic observation over time, it was found that within a week, a number of fibroblasts gathered around the AVNCs and formed cell clusters, and thereafter the cell clusters started to beat spontaneously. The nascent cell clusters expanded their area gradually by three weeks in culture and expressed specific cardiac genes and proteins. Maturation of newly formed cardiomyocytes seems to be slow in cultures of AVNCs compared with those of sinoatrial node cells. Stimulation of muscarinic receptors with acetylcholine induced a beating rate decrease which was blocked by atropine, and activation of adenylate cyclase activity with forskolin increased the beat rate, while stimulation of beta adrenoceptors by isoproterenol had no effect. These results indicate that AVNCs form a cluster of cells with properties of functional cardiomyocytes and provide evidence to support the hypothesis.     

Nonlinear modeling of the atrioventricular node physiology in atrial fibrillation

A nonlinear model of the atrioventricular (AV) node physiology in atrial fibrillation (AF) is proposed based on three assumptions: (1) normal distribution of atrial impulses, (2) right-skewed distribution of R-R intervals, (3) increase in the refractory period of the AV node due to rapid bombardment from the atria. Simulation resulted in the following conclusions, all of which are in agreement with previous experience: (1) the entry speed of atrial impulses into the AV node in AF is inversely proportional to the ventricular rate, (2) the autocorrelation function of R-R intervals is zero at all delays, (3) a newly introduced index, sign of first difference, has a negative autocorrelation function at the first delay and zero ones at all others. In spite of its simplicity, the model is able to explain what happens in atrial premature complexes, sinus tachycardia and sinus bradycardia. Different rhythms, some of which rarely seen clinically, can be reproduced by changing input patterns or by slightly manipulating the model parameters. In order to make possible a long irregular time series of R-R interval, aperiodic changes in atrial signals are shown to be necessary. In conclusion, we proposed a simple model for the AV node physiology capable of explaining the previously known facts about AF as well as predicting interesting properties of some other supraventricular arrhythmias.     

Modulation by endothelin-1 of spontaneous activity and membrane currents of atrioventricular node myocytes from the rabbit heart

Background

The atrioventricular node (AVN) is a key component of the cardiac pacemaker-conduction system. Although it is known that receptors for the peptide hormone endothelin-1 (ET-1) are expressed in the AVN, there is very little information available on the modulatory effects of ET-1 on AVN electrophysiology. This study characterises for the first time acute modulatory effects of ET-1 on AVN cellular electrophysiology.

Methods

Electrophysiological experiments were conducted in which recordings were made from rabbit isolated AVN cells at 35–37°C using the whole-cell patch clamp recording technique.

Results

Application of ET-1 (10 nM) to spontaneously active AVN cells led rapidly (within ∼13 s) to membrane potential hyperpolarisation and cessation of spontaneous action potentials (APs). This effect was prevented by pre-application of the ET_A receptor inhibitor BQ-123 (1 µM) and was not mimicked by the ET_B receptor agonist IRL-1620 (300 nM). In whole-cell voltage-clamp experiments, ET-1 partially inhibited L-type calcium current (I_Ca,L) and rapid delayed rectifier K⁺ current (I_Kr), whilst it transiently activated the hyperpolarisation-activated current (I_f) at voltages negative to the pacemaking range, and activated an inwardly rectifying current that was inhibited by both tertiapin-Q (300 nM) and Ba²⁺ ions (2 mM); each of these effects was sensitive to ET_A receptor inhibition. In cells exposed to tertiapin-Q, ET-1 application did not produce membrane potential hyperpolarisation or immediate cessation of spontaneous activity; instead, there was a progressive decline in AP amplitude and depolarisation of maximum diastolic potential.

Conclusions

Acutely applied ET-1 exerts a direct modulatory effect on AVN cell electrophysiology. The dominant effect of ET-1 in this study was activation of a tertiapin-Q sensitive inwardly rectifying K⁺ current via ET_A receptors, which led rapidly to cell quiescence.     

Pacemaker activity and ionic currents in mouse atrioventricular node cells

It is well established that Pacemaker activity of the sino-atrial node (SAN) initiates the heartbeat. However, the atrioventricular node (AVN) can generate viable pacemaker activity in case of SAN failure, but we have limited knowledge of the ionic bases of AVN automaticity. We characterized pacemaker activity and ionic currents in automatic myocytes of the mouse AVN. Pacemaking of AVN cells (AVNCs) was lower than that of SAN pacemaker cells (SANCs), both in control conditions and upon perfusion of isoproterenol (ISO). Block of I(Na) by tetrodotoxin (TTX) or of I(Ca,L) by isradipine abolished AVNCs pacemaker activity. TTX-resistant (I(Nar)) and TTX-sensitive (I(Nas)) Na(+) currents were recorded in mouse AVNCs, as well as T-(I(Ca,T)) and L-type (I(Ca,L)) Ca(2+) currents I(Ca,L) density was lower than in SANCs (51%). The density of the hyperpolarization-activated current, (I(f)) and that of the fast component of the delayed rectifier current (I(Kr)) were, respectively, lower (52%) and higher (53%) in AVNCs than in SANCs. Pharmacological inhibition of I(f) by 3 μM ZD-7228 reduced pacemaker activity by 16%, suggesting a relevant role for I(f) in AVNCs automaticity. Some AVNCs expressed also moderate densities of the transient outward K(+) current (I(to)). In contrast, no detectable slow component of the delayed rectifier current (I(Ks)) could be recorded in AVNCs. The lower densities of I(f) and I(Ca,L), as well as higher expression of I(Kr) in AVNCs than in SANCs may contribute to the intrinsically slower AVNCs pacemaking than that of SANCs.     

Laser endoscopic destruction of the atrioventricular junction of the heart conduction system in an experiment

Feasibility and degree of local AV junction destruction were studied prior to the induction of complete transversal block on the model of 18 isolated perfused dog hearts, using endoscopic laser device, employing cardiofibroscope and laser Ne: YAG. Morphological pattern in the destruction zone is presented. The technique and device can be used for transvenous AV junction destruction.     

The HAV pattern in pediatric patients with atrioventricular node reentrant tachycardia

ObjectivesThe purpose of this study is to assess the prevalence of a His-Atrial-Ventricular (HAV) pattern, i.e. the atrial electrogram following the His bundle -HB- electrogram and preceding the ventricular one, on the catheter placed in the His position in pediatric patients during typical atrioventricular node reentry (AVNRT).Materials and methodsThe pediatric electrophysiology databases of two separate institutions were queried for patients with a diagnosis of AVNRT. Demographic, clinical data and the electrophysiology study (EPS) information were assessed.ResultsThirty-nine consecutive patients were included. Twenty-five were female. The average age at the time of the EPS was 12 ± 3.7 years. Induction was achieved with atrial pacing in 23, with a single atrial extra stimulus in 8 and with dual atrial extra stimuli in 8. Isoproterenol was needed to induce tachycardia in 21. Tachycardia cycle length averaged 320 ± 50 ms. An HAV pattern was present in 35 (74%) of the patients, and in 100% of the patients younger than 8.ConclusionsAn HAV pattern on the catheter placed in the His position, is common in pediatric patients with AVNRT, occurring in up to 74% of the patients in this population, being more common in younger patients.     

DNA-based identification of a hepatic trematode in an elk calf

Liver fluke infection was identified as a probable cause of clinical disease in an approximately 6-mo-old elk (Cervus elaphus) in coastal Oregon. Clinical pathology and necropsy findings are described. The alcohol-fixed flukes that were submitted for identification were similar in size to Fasciola hepatica, but their shape resembled Fascioloides magna in that they lacked a distinctive anterior cone. A few structures consistent with the eggs of F. magna were observed in liver lesions, suggesting that at least some of the worms were sexually maturing. Due to difficulties in morphologic identification associated with improper fixation technique, DNA analysis was used to compare small subunit (SSU) and internal transcribed spacer 2 ribosomal RNA gene sequences of the recovered parasites with those of F. hepatica and F. magna, confirming these small, but sexually mature flukes were F. magna. This is the first publication of the SSU gene sequence for F. magna. Phylogenetic analysis showed that it is related to, but is an outlier, to the genus Fasciola. Due to the high mortality rate associated with this disease outbreak, the overall significance of trematodiasis in the herd is unclear.