Ultrastructural study of the myocardium of the sinus venosus and sinoatrial valve in the dogfish (Scyliorhinus canicula)

The myocardium of the sinus venosus of the dogfish ( Scyliorhinus canicula ) is located between a thick subepicardial collagen-rich layer and a subendocardial network of nerve fibres and ganglion cells. The sinoatrial valve consists of two transversal folds of the cardiac wall which are separated by connective tissue, except in their free margins.
The myocardium of the sinus venosus and the sinusal face of the sinoatrial valve is arranged in bundles which are surrounded by a 40 nm-thick basal lamina. The myocardial cells measure about 7-9 μm in diameter at the nuclear level. Nerve terminals are frequent in the centre of the bundles. Most of the sinusal myocardiocytes have a scarce amount of myofibrils which are randomly orientated. The sarcoplasmic reticulum is relatively well developed and consists of peripheral couplings, subsarcolemmal vesicles, circular and longitudinal tubules. The scarce intercalated discs show only fasciae adhaerentes . Gap junctions, desmosomes or specific granules are not observed in the sinusal myocardiocytes of the dogfish. In contrast, the atrial myocardiocytes are smaller, about 5-6 μm in diameter at the nuclear level. The cytoplasm is denser and the myofibrils are abundant and orientated in parallel directions. Specific granules are present. although scarce. Subsarcolemmal vesicles are less frequent, while the atrial intercalated discs are larger and more abundant than those of the sinus venosus. Neural elements are scarce in the atrium.
The differences observed between sinus venosus and atrium might be related to the morphological criteria to distinguish between the nodal tissue and the working myocardiocytes of higher vertebrates. On the other hand, we think that the connective tissue placed between sinus venom and atrium means that the contraction impulse generated in the sinus venosus must reach the atrium through the free margin of the valve. It might play a role in the sinoatrial valve function.     

Endocardial pores in the sinus venosus and atrium of the dogfish

A scanning electron microscopy study showed the presence of large pores in the endocardium of the sinus venosus of the dogfish Scyliorhinus canicula. The pores were always found on large bundles which protruded into the cardiac lumen. The bundles were mainly constituted of granule-containing nerve fibres. The average diameter of the pores was 3.2 μm (range=1.5-5.0μm), and their density, was about 822 pores mm⁻². Endocardial pores were absent in other areas of the sinus venosus, but they were observed on the little bundles of granulated nerve fibres which were scattered throughout the atrium. The existence of large endocardial pores associated with bundles of granulated nerve fibres supports the hypothesis for the neuroendocrine nature of the elasmobranch sinus venosus wall.     

Heart inflow tract of the African lungfish Protopterus dolloi

We report a morphologic study of the heart inflow tract of the African lungfish Protopterus dolloi. Attention was paid to the atrium, the sinus venosus, the pulmonary vein, and the atrioventricular (AV) plug, and to the relationships between all these structures. The atrium is divided caudally into two lobes, has a common part above the sinus venosus, and appears attached to the dorsal wall of the ventricle and outflow tract through connective tissue covered by the visceral pericardium. The pulmonary vein enters the sinus venosus and runs longitudinally toward the AV plug. Then it fuses with the pulmonalis fold and disappears as an anatomic entity. However, the oxygenated blood is directly conveyed into the left atrium by the formation of a pulmonary channel. This channel is formed cranially by the pulmonalis fold, ventrally by the AV plug, and caudally and dorsally by the atrial wall. The pulmonalis fold appears as a wide membranous fold which arises from the left side of the AV plug and extends dorsally to form the roof of the pulmonary channel. The pulmonalis fold also forms the right side of the pulmonary channel and sequesters the upper left corner of the sinus venosus from the main circulatory return. The AV plug is a large structure, firmly attached to the ventricular septum, which contains a hyaline cartilaginous core surrounded by connective tissue. The atrium is partially divided into two chambers by the presence of numerous pectinate muscles extended between the dorsal wall of the atrium and the roof of the pulmonary channel. Thus, partial atrial division is both internal and external, precluding the more complete division seen in amphibians. The present report, our own unpublished observations on other Protopterus, and a survey of the literature indicate that not only the Protopterus, but also other lungfish share many morphologic traits.     

Polar effects of lithium in the heart of the zebrafish Danio rerio

 The molecular signalling mechanisms that are believed to govern the patterning of the heart early in embryonic development are not well understood. We have investigated the events which occur during patterning of the vertebrate heart by exposing gastrula stage zebrafish embryos to lithium, which is known to affect the phosphoinositol signalling pathway. Treatment of embryos at 50% epiboly (5.25 h after fertilization at 28.5°C) with 0.3 m LiCl for 5–15 min, results in embryos with defects which range from mild to severe, depending on the length of time the embryos are exposed to lithium. In the heart, defects appear progressively in the inflow tract, the sinus venosus and atrium. By using an antibody that recognizes an atrium-specific isoform of myosin, our results show that lithium treatment at gastrulation specifically affects the atrium and sinus venosus, and has little obvious effect on the ventricle. Defects induced by lithium differ from those induced by retinoic acid (RA) treatment of similarly staged embryos, and suggest that lithium and RA may affect the patterning signals important for establishment of the vertebrate heart by acting on different populations of cells or by influencing different patterning pathways. Received: 8 December 1995 / Accepted: 11 April 1996     

Morpho-functional characterization of the goldfish (Carassius auratus L.) heart

Using morphological and physiological approaches we provided, for the first time, a structural and functional characterization of Carassius auratus L. heart. Besides to the classical four chambers, i.e. sinus venosus, atrium, ventricle, bulbus, we described two distinct structures corresponding to the atrio-ventricular (AV) region and the conus arteriosus. The atrium is very large and highly trabeculated; the ventricle shows an outer compacta, vascularized by coronary vessels, and an inner spongiosa; the bulbus wall is characterized by a high elastin/collagen ratio, which makes it extremely compliant. Immunolocalization revealed a strong expression of activated "eNOS-like" isoforms both at coronary endothelium and, to a lesser extent, in the myocardiocytes and the endocardial endothelium (EE). The structural design of the heart appears to comply with its mechanical function. Using an in vitro working heart preparation, cardiac performance was evaluated at different filling and afterload pressures. The hearts were very sensitive to filling pressure increases. Maximum Stroke volume (SV=1.08 ± 0.09 mL/kg body mass) was obtained with an input pressure of 0.4 kPa. The heart was not able to sustain afterload increases, values higher than 1.5 kPa impairing its performance. These morpho-functional features are consistent with a volume pump mechanical performance.     

The Electrophysiological Organization of the Embryonic Chick Heart

Both intracellular and surface electrodes were employed to record electrical activity from embryonic chick hearts between the ages of 3 and 20 days. Cells from the sinus venosus, sinoatrial (SA) valves, atrium, atrioventricular (AV) ring, and ventricle were localized and characterized on the basis of shape, amplitude, rise time, and duration of transmembrane potentials. The differences in transmembrane potentials from these various regions provided the basis for a hypothesis concerned with the distribution of pacemaker potentiality and one related to the origin of the His-Purkinje system. Action potentials recorded along the entire embryonic AV ring were comparable to those of the adult rabbit AV nodal cells in both configuration and sequence of activation and were thus categorized into three functional regions (AN, N, NH). Histological sections of 7 and 14 day hearts demonstrated muscular continuity between the right atrium and ventricle across the muscular AV valve.     

A radiological study of the central circulation in the lungfish, Protopterus aethiopicus

The dipnoan heart is only in part structurally developed to support a separated circulation in pulmonary and systemic circuits. In the present investigation biplane angiocardiography has been used to describe the extent of such a double circulation and the factors which may modify it in the African lungfish, Protopterus aethiopicus. Contrast injections in the pulmonary vein revealed a clear tendency for aerated blood returing from the lungs to be selectively dispatched to the anterior branchial arteries giving rise to the major systemic circulation. Contrast injections in the vena cava delineated the sinus venosus as a large receiving chamber for systemic venous blood. Contraction of the sinus venosus discharged blood into the right, posterior part of the partially divided atrial space. Contrast injection in the pulmonary vein showed that vessel to pass obliquely from right to left such that blood was emptied distinctly into the left side of the atrium. During contraction the atrial space tended to retain a residual volume in its anterior undivided part which minized mixing. Ventricular filling occurred through separate right and left atrio-ventricular connections. Right-left separation in most of the ventricle was maintained by the partial ventricular septum, the trabeculated, spongelike myocardium and the mode of inflow from the atria. Mixing in the anterior undivided portion of the ventricle during the ejection phase was slight due to a streamlined ejection pattern. The outflow through the bulbus cordis occurred in discrete streams which in part were structurally separated by well developed spiral folds. In the anterior bulbus segment the spiral folds are fused and make completely separate dorsal and ventral outflow tracts. The ventral bulbus channel provides blood to the three anterior branchial arteries. The second and third branchial arteries are large and represent direct shunts to the dorsal aorta. The fourth and fifth branchial arteries are gill bearing and receive blood form the dorsal bulbus channel. The most posterior epibranchial vessels give rise to the pulmonary arteries.     

An Angiocardiographic Analysis of the Central Circulation in the Air Breathing Teleost Channa argus

The unique anatomy of the double ventral aorta outflow system in the air breathing teleost Channa argus (Ophiocephalus) showing an anterior and posterior ventral aorta is described. The marked trabeculation of the ventricle and bulbus arteriosus and the arrangement of central veins are used as a basis for the hypothesis that Channa may selectively channel the well oxygenated blood draining the air breathing organs via the anterior cardinal vein to the posterior ventral aorta, which forms the systemic arterial circulation. An angiocardiographic technique was used to test this hypothesis, as well as to delineate the functional role of the heart chambers in the cardiac cycle. No reflux of contrast to the sinus venosus during atrial filling and no ventricular filling before atrial contraction were apparent, which makes the atrium the main determinant of the ventricular end-diastolic volume. Ventricular contraction left a small or no residual volume. The ventricular ejectate was initially nearly completely absorbed by the very elastic bulbus arteriosus, acting as a pressure chamber (Windkessel) stabilizing and prolonging ventral aortic blood flow. Contrast medium was not selectively passed from the anterior cardinal vein to the posterior ventral aorta. However, the diameter of this vessel and its density of contrast were greater than in the anterior aorta, suggesting a preference for a greater blood flow from the air breathing organ through the heart to the posterior aorta.     

Identification and detection of the isolated sinus venosus from the Asian toad

The pacemaker activity of mammalian sinoatrial node (SAN) of the heart plays a fundamental role in the integration of vital functions. Studying factors such as drugs that influence pacemaker activity of SAN has its significance. In this study, we isolated sinus venosus, SAN from toads (Bufo gargarizans), and analysed its electronic signal, histological characteristics and the influence of acetylcholine (ACh) and ivabradine on its pacemaker activity using PowerLab® and Chart® 5.0 software. We found that when isolated sinus venosus was treated with ACh, its histological distribution was disorganized and inter‐beat (RR) interval was also broadened. The high frequency normalized unit (HFnu) and Poincaré plot of heart rate variability (HRV) of the isolated sinus venosus was also altered upon ACh treatment in a time‐dependent and dose‐dependent manner. When treated with ivabradine, these parameters of HRV such as square root of the mean of the squared differences between adjacent NN intervals (RMSSD) and HFnu were in the upward tendency, but low frequency normalized unit and low frequency/high frequency were in the opposite tendency. Taken together, we have developed a new model for studying the influences of drugs on autorhythmicity using isolated sinus venosus of the toad. With this model, we showed that ACh and ivabradine may affect the pacemaker activity by stimulating muscarinic receptor or inhibiting I_f current, respectively. Copyright © 2013 John Wiley & Sons, Ltd.     

Functional Morphology of the Heart in Fishes

The systemic heart of fishes consists of four chambers in series,the sinus venosus, atrium, ventricle, and conus or bulbus. Valvesbetween the chambers and contraction of all chambers exceptthe bulbus maintain a unidirectional blood flow through theheart. The heart is composed of typical vertebrate cardiac muscle,although there may be minor differences in the distributionof spontaneously active cells, the rate and nature of spreadof excitatory waves, and the characteristics of resting andaction potentials between different fish and other vertebrates.Cholinergic fibers innervate the heart, except in hagfish whichhave aneural hearts. Fish hearts lack sympathetic innervation.The level of vagal tone varies considerably, and is affectedby many factors. In some fish the heart is essentially aneural(without vagal tone) during exercise and may resemble an isolatedmammalian ventricle with increased venous return causing increasedcardiac output. There are many mechanisms that could increasevenous return in exercising fish. rß-adrenergic receptorshave been located on the hearts of some fish, and changing levelsof catecholamines may play a role in regulating cardiac activity.Changes in cardiac output in fish are normally associated withlarge changes in stroke volume and small cha-nges in heart rate.     

Cardiac Function in the Hagfish,Myxine (Myxinoidea:Cyclostomata)

The electrocardiogram of Myxine glutinosa is described and its events related to systole in the sinus venosus, atrium and ventricle. Intraatrial, ventricular, ventral aortic and dorsal aortic pressures are presented. Ventricular systolic pressure was 10.3 ± 2.0 cm H₂O; average dorsal aortic pressure was 7.0 ± 1.7 cm H₂O. The resistance of the gill vessels caused a loss of 28% of the ventral aortic pressure. The conclusion is drawn that the basic mechanism of the heart of Myxine is like that of fish and higher vertebrates, and that the low pressure it develops is to be ascribed to structural and functional features of the myocardium.     

Syngnathinema californiense n. gen., n. sp. (Nematoda: Dracunculoidea) from the circulatory system of the bay pipefish Syngnathus leptorhynchus in California.

A new dracunculoid nematode Syngnathinema californiense, n. gen., n. sp., is described from gravid female parasites found inside the heart of wild-caught Bay pipefish Syngnathus leptorhynchus; histologically it was recorded from different sites of the host circulatory system (sinus venosus, atrium, and renal and hepatic veins). The new genus is characterized by the structure of the cephalic end (large oral aperture with a narrow peribuccal ring surrounded by 10 papillae in 2 circles, large circular amphids), division of the esophagus into muscular and glandular portions, location of the well-developed vulva posterior to the esophagus, monodelphic female reproductive organs, and the sharply pointed tail. Because males remain unknown, the genus is provisionally assigned to the Daniconematidae.     

On the ultrastructure of the sinus venosus in Chimara monstrosa L. (Elasmobranchii: Holocephali)

Summary The wall of the sinus venosus in an elasmobranchian species, Chimaera monstrosa L. is described.Endocardial cells contain numerous large vacuoles, as well as a number of membrane-bounded, moderately electron dense bodies (MDB). Myocardial cells lie closely packed into bundles surrounded by a basal lamina of about 20 nm thickness, and by large amounts of collagen fibres. These cells are connected by desmosomes of 1–2 µm length and with an intermembranous gap of 10–20 nm. Myocardial cells poor in myofibrils are intermingled with cells containing a well developed contractile material. Atrial specific granules are scarce. Vesiculated nerve processes occur at a distance of about 20 nm from the myocardial sarcolemma. Myocardial cells of the sino-atrial junction appear ultrastructurally similar to those located elsewhere in the sinus venosus. Epicardial cells contain large vacuoles, and have fibrecoated protrusions extending into the pericardial space.The possibility of pacemaker activity in the elasmobranchian sinus venosus is discussed.     

A comparative study of the adrenergic innervation of the teleost heart

Summary An adrenergic cardiac innervation has been found in the following teleost species:Platycephalus bassensis (Platycephalidae);Atopomycterus nicthemerus (Diodontidae);Aracana ornata (Ostraciontidae);Torquiginer glaber (Tetraodontidae);Aldrichetta forsteri (Mugilidae);Anguilla australis occidentalis (Anguillidae). In contrast, no evidence for an adrenergic cardiac innervation was found in the pleuronectid,Rhombosolea tapirina.Fluorescence histochemical studies indicated that adrenergic nerves in the sinus venosus and atrium entered via the vagus, whereas those to the ventricle passed mainly along the coronary vasculature. No fluorescent nerves were observed in the heart ofRhombosolea.Transmural stimulation of these adrenergic nerves increased the force of beat in the atrium and ventricle ofPlatycephalus, Atopomycterus andAracana, and in the atrium only ofTorquiginer, Aldrichetta andAnguilla. In addition stimulation of the abdominal vagus nerve in the presence of hyoscine increased the force of beat and heart rate in spontaneously beating sinus-atrium preparations of all species exceptRhombosolea. Applied catecholamines increased the force of beat and heart rate in the spontaneously beating heart of all species includingRhombosolea.     

The cardiovascular chromaffin cell system of the southern hemisphere lamprey,Geotria australis gray

This study demonstrates that the silver technique of Grimelius (Acta Soc. Med. Ups. 73:243–270, 68) is ideally suited for the study of cardiovascular chromaffin cells in lampreys. This method showed that in the Southern Hemisphere lamprey, Geotria australis, the distribution of chromaffin cells differs from that described for holarctic species. In G. australis, the chromaffin cells are found mainly in the sinus venosus, atrium, and nearby regions of the cardinal and jugular veins, and they are absent from the ventricle and conus arteriosus. The location and discreteness of the large accumulation of chromaffin cells in the lateral wall of the right posterior cardinal vein of adults resemble those of the precardiac axillary bodies of elasmobranchs. Chromaffin cells become more abundant during metamorphosis. The possible phylogenetic and functional significance of lamprey chromaffin cells is briefly discussed.     

Subepicardial endothelial cells invade the embryonic ventricle wall to form coronary arteries

Coronary arteries bring blood flow to the heart muscle. Understanding the developmental program of the coronary arteries provides insights into the treatment of coronary artery diseases. Multiple sources have been described as contributing to coronary arteries including the proepicardium, sinus venosus (SV), and endocardium. However, the developmental origins of coronary vessels are still under intense study. We have produced a new genetic tool for studying coronary development, an AplnCreER mouse line, which expresses an inducible Cre recombinase specifically in developing coronary vessels. Quantitative analysis of coronary development and timed induction of AplnCreER fate tracing showed that the progenies of subepicardial endothelial cells (ECs) both invade the compact myocardium to form coronary arteries and remain on the surface to produce veins. We found that these subepicardial ECs are the major sources of intramyocardial coronary vessels in the developing heart. In vitro explant assays indicate that the majority of these subepicardial ECs arise from endocardium of the SV and atrium, but not from ventricular endocardium. Clonal analysis of Apln-positive cells indicates that a single subepicardial EC contributes equally to both coronary arteries and veins. Collectively, these data suggested that subepicardial ECs are the major source of intramyocardial coronary arteries in the ventricle wall, and that coronary arteries and veins have a common origin in the developing heart.     

The sinus venosus myocardium contributes to the atrioventricular canal: potential role during atrioventricular node development?

The presence of distinct electrophysiological pathways within the atrioventricular node (AVN) is a prerequisite for atrioventricular nodal reentrant tachycardia to occur. In this study, the different cell contributions that may account for the anatomical and functional heterogeneity of the AVN were investigated. To study the temporal development of the AVN, the expression pattern of ISL1, expressed in cardiac progenitor cells, was studied in sequential stages performing co‐staining with myocardial markers (TNNI2 and NKX2‐5) and HCN4 (cardiac conduction system marker). An ISL1+/TNNI2+/HCN4+ continuity between the myocardium of the sinus venosus and atrioventricular canal was identified in the region of the putative AVN, which showed a pacemaker‐like phenotype based on single cell patch‐clamp experiments. Furthermore, qPCR analysis showed that even during early development, different cell populations can be identified in the region of the putative AVN. Fate mapping was performed by in ovo vital dye microinjection. Embryos were harvested and analysed 24 and 48 hrs post‐injection. These experiments showed incorporation of sinus venosus myocardium in the posterior region of the atrioventricular canal. The myocardium of the sinus venosus contributes to the atrioventricular canal. It is postulated that the myocardium of the sinus venosus contributes to nodal extensions or transitional cells of the AVN since these cells are located in the posterior region of the AVN. This finding may help to understand the origin of atrioventricular nodal reentrant tachycardia.     

Tachykinin-like immunoreactivity in the sinus venosus of the dogfish (Scyliorhinus canicula)

The sinus venosus of the elasmobranch heart is characterized by the presence of large bundles of unmyelinated nerve fibres that bulge into the cardiac lumen, below the endocardium. In the dogfish (Scyliorhinus canicula), these fibres contain numerous dense-core membrane-bounded granules of about 200 nm in diameter. Most intramural ganglion cells of the sinus venosus also show densely packed granules similar to those found in the subendocardial fibres. We have observed strong substance-P-like immunoreactivity in the large fibre bundles and in the perikarya of the ganglion cells. Preabsorption of the antisera with fragment 7–11 of substance P has shown that the antisera recognize the tachykinin canonic sequence. Our findings suggest that an undetermined tachykinin is secreted in the elasmobranch heart, and that it is probably released into the blood stream in the context of a little-known neuroendocrine system.     

Morphological analysis of the hagfish heart. II. The venous pole and the pericardium

The morphological characteristics of the venous pole and pericardium of the heart were examined in three hagfish species, Myxine glutinosa, Eptatretus stoutii, and Eptatretus cirrhatus. In these species, the atrioventricular (AV) canal is long, funnel‐shaped and contains small amounts of myocardium. The AV valve is formed by two pocket‐like leaflets that lack a papillary system. The atrial wall is formed by interconnected muscle trabeculae and a well‐defined collagenous system. The sinus venosus (SV) shows a collagenous wall and is connected to the left side of the atrium. An abrupt collagen‐muscle boundary marks the SV‐atrium transition. It is hypothesized that the SV is not homologous to that of other vertebrates which could have important implications for understanding heart evolution. In M. glutinosa and E. stoutii, the pericardium is a closed bag that hangs from the tissues dorsal to the heart and encloses both the heart and the ventral aorta. In contrast, the pericardium is continuous with the loose periaortic tissue in E. cirrhatus. In all three species, the pericardium ends at the level of the SV excluding most of the atrium from the pericardial cavity. In M. glutinosa and E. stoutii, connective bridges extend between the base of the aorta and the ventricular wall. In E. cirrhatus, the connections between the periaortic tissue and the ventricle may carry blood vessels that reach the ventricular base. A further difference specific to E. cirrhatus is that the adipose tissue associated with the pericardium contains thyroid follicles. J. Morphol. 277:853–865, 2016. © 2016 Wiley Periodicals, Inc.