Pigmentation of the aortic and pulmonary valves in C57BL/6J x Balb/cByJ hybrid mice of different coat colours.

Neural crest-derived melanocytes have been recorded in several parts of the mammalian heart but not in the pulmonary valve. We report here the presence of melanin-containing cells in the leaflets (cusps) of both the aortic and pulmonary valves. A total of 158 C57BL/6J x Balb/cByJ hybrid mice exhibiting four coat colours, namely black, white, agouti and non-agouti brown, were examined. We sought for any relationship between the presence of melanocytes in the valves and the coat colour of the animals. The pigmentation levels of the leaflets were accomplished using a scale of five pigment intensities. White mice lacked pigment in the heart. In 10.5% of the remaining animals, there were melanocytes in the pulmonary valve leaflets. Thus, this is the first study to report the presence of such cells in the pulmonary valve of mammals. Melanocytes occurred in the leaflets of the aortic valves of 87.2% of mice. The incidence of melanocytes and the pigmentation level of the leaflets did not statistically differ according to the coat colours of the animals. This disagrees with previous observations, indicating that the amount of melanocytes in the heart reflects that of the skin. The incidence and distribution of melanocytes in aortic and pulmonary valves are consistent with the notion that the formation of the arterial valves is mediated by specific subpopulations of neural crest cells. We hypothesize that melanocytes, even not producing melanin, may be more frequent in the heart than previously thought, exerting presumably an immunological function.

The valves of the cardiac outflow tract of the starry ray, Raja asterias (Chondrichthyes; Rajiformes): Anatomical, histological and evolutionary aspects.

The cardiac outflow tract of chondrichthyans is composed of the myocardial conus arteriosus, equipped with valves at its luminal side, and the bulbus arteriosus devoid of myocardium. Knowledge of the histomorphology of the conal valves is scarce despite their importance in preventing blood backflow to the heart. Current information on the subject refers to a single shark species. The present report is the first to describe the structure of the conal valves of a batoid species, namely, Raja asterias. Hearts from seven starry rays were examined using scanning electron microscopy and histochemical techniques for light microscopy. In all hearts, the conus showed four transverse rows of three pocket-like valves each. Each valve was composed of a leaflet and its supporting sinus. The leaflet had a stout central body, rich in glycosaminoglycans, which contained fibroblasts, collagen and elastin. The central body was surrounded by two thin fibrous layers, outer and inner, formed mainly by collagen. The valves of the anterior row, which were the largest of the valvular system, were attached proximally to the conus arteriosus and distally to the bulbus arteriosus, and not to the ventral aorta as previously reported for chondrichthyans. The arrangement of the anterior valves in the starry ray is an anatomical pattern that apparently has been preserved throughout the evolution of vertebrates.

The anatomical components of the cardiac outflow tract of chondrichthyans and actinopterygians.

The outflow tract of the fish heart is the segment interposed between the ventricle and the ventral aorta. It holds the valves that prevent blood backflow from the gill vasculature to the ventricle. The anatomical composition, histological structure and evolutionary changes in the fish cardiac outflow tract have been under discussion for nearly two centuries and are still subject to debate. This paper offers a brief historical review of the main conceptions about the cardiac outflow tract components of chondrichthyans (cartilaginous fish) and actinopterygians (ray-finned fish) which have been put forward since the beginning of the nineteenth century up to the current day. We focus on the evolutionary origin of the outflow tract components and the changes to which they have been subject in the major extant groups of chondrichthyans and actinopterygians. In addition, an attempt is made to infer the primitive anatomical design of the heart of the gnathostomes (jawed vertebrates). Finally, several areas of further investigation are suggested. Recent work on fish heart morphology has shown that the cardiac outflow tract of chondrichthyans does not consist exclusively of the myocardial conus arteriosus as classically thought. A conus arteriosus and a bulbus arteriosus, devoid of myocardium and mainly composed of elastin and smooth muscle, are usually present in cartilaginous and ray-finned fish. This is consistent with the suggestion that both components coexisted from the onset of the gnathostome radiation. There is evidence that the conus arteriosus appeared in the agnathans. By contrast, the evolutionary origin of the bulbus is still unclear. It is almost certain that in all fish, both the conus and bulbus develop from the embryonic second heart field. We suggest herein that the primitive anatomical heart of the jawed vertebrates consisted of a sinus venosus containing the pacemaker tissue, an atrium possessing trabeculated myocardium, an atrioventricular region with compact myocardium which supported the atrioventricular valves, a ventricle composed of mixed myocardium, and an outflow tract consisting of a conus arteriosus, with compact myocardium in its wall and valves at its luminal side, and a non-myocardial bulbus arteriosus that connected the conus with the ventral aorta. Chondrichthyans have retained this basic anatomical design of the heart. In actinopterygians, the heart has been subject to notable changes during evolution. Among them, the following two should be highlighted: (i) a decrease in size of the conus in combination with a remarkable development of the bulbus, especially in teleosts; and (ii) loss of the myocardial compact layer of the ventricle in many teleost species.

The bulbus arteriosus of the holocephalan heart: gross anatomy, histomorphology, pigmentation, and evolutionary significance.

This study was designed to determine whether the outflow tract of the holocephalan heart is composed of a myocardial conus arteriosus and a non-myocardial bulbus arteriosus, as is the case in elasmobranchs. This is a key issue to verify the hypothesis that these two anatomical components existed from the onset of the jawed vertebrate radiation. The Holocephali are the sister group of the elasmobranchs, sharing with them a common, still unknown Palaeozoic ancestor. The sample examined herein consisted of hearts from individuals of four species, two of them belonging to the Chimaeridae and the other two to the Rhinochimaeridae. In all specimens, the cardiac outflow tract consisted of a conus arteriosus, with myocardium in its walls and two rows of valves at its luminal side, and an intrapericardial bulbus arteriosus shorter than the conus and devoid of valves. The bulbus, mainly composed of elastin and smooth musculature, was covered by the epicardium and crossed longitudinally by coronary artery trunks. These findings give added support to the viewpoint that the outflow tract of the primitive heart of the gnathostomes was not composed of a single component, but two, the conus and the bulbus. All rabbitfish (Chimaera monstrosa) examined had pigment cells over the surface of the heart. The degree of pigmentation, which varied widely between individuals, was particularly intense in the cardiac outflow tract. Pigment cells also occurred in the bulbus arteriosus of one of the two hearts of the straightnose rabbitfish (Rhinochimaera atlantica) included in the study. The cells containing pigment, presumably derived from the neural crest, were located in the subepicardium.

Anatomical, histochemical and immunohistochemical characterisation of the cardiac outflow tract of the silver arowana, Osteoglossum bicirrhosum (Teleostei: Osteoglossiformes).

The cardiac outflow tract of chondrichthyans and actinopterygians is composed of a myocardial conus arteriosus and a non-myocardial bulbus arteriosus. In teleosts, the conus has been subjected to a reduction in size over the evolution in conjunction with the further development of the bulbus. Most studies on the outflow tract of the teleost heart refer to species of modern groups and are mainly devoted to the bulbus. Knowledge on the outflow tract of species belonging to early teleost groups is scarce. The aim here was to characterise the structure of the cardiac outflow tract of the silver arowana, a representative of the ancient teleost clade of the Osteoglossomorpha. The material consisted of hearts from six juvenile animals. The cardiac outflow tract of the silver arowana is composed of a conus, which supports two conal valves, and a bulbus. Both components are lined externally by the epicardium and internally by the endocardium. The conus is immunoreactive to antibodies against myosin heavy chains and is composed of compact myocardium, thus contrasting with the ventricle, which has exclusively trabeculated myocardium. The bulbus is immunoreactive to antibodies against smooth muscle α-actin and mainly consists of elastic fibres and smooth muscle cells, both arranged in three layers, outer, middle and inner. The most remarkable feature of the bulbus is the presence of two prominent longitudinal ridges, dorsal and ventral, at the luminal side, which serve to anchor the commissures of the conal valves. This arrangement has not been described so far in any fish species. Pigment cells, presumably of neural crest origin, are present in the subepicardium of the bulbus and anterior part of the ventricle.

Structure and vascularization of the ventricular myocardium in Holocephali: their evolutionary significance.

It was generally assumed that the ventricle of the primitive vertebrate heart was composed of trabeculated, or spongy, myocardium, supplied by oxygen-poor luminal blood. In addition, it was presumed that the mixed ventricular myocardium, consisting of a compacta and a spongiosa, and its supply through coronary arteries appeared several times throughout fish evolution. Recent work has suggested, however, that a fully vascularized, mixed myocardium may be the primitive condition in gnathostomes. The present study of the heart ventricles of four holocephalan species aimed to clarify this controversy. Our observations showed that the ventricular myocardium of Chimaera monstrosa and Harriotta raleighana consists of a very thin compacta overlying a widespread spongiosa. The ventricle of Hydrolagus affinis is composed exclusively of trabeculated myocardium. In these three species there is a well-developed coronary artery system. The main coronary artery trunks run along the outflow tract, giving off subepicardial ventricular arteries. The trabeculae of the spongiosa are irrigated by branches of the subepicardial arteries and by penetrating arterial vessels arising directly from the main coronary trunks at the level of the conoventricular junction. The ventricle of Rhinochimaera atlantica has only spongy myocardium supplied by luminal blood. Small coronary arterial vessels are present in the subepicardium, but they do not enter the myocardial trabeculae. The present findings show for the first time that in a wild living vertebrate species, specifically H. affinis, an extensive coronary artery system supplying the whole cardiac ventricle exists in the absence of a well-developed compact ventricular myocardium. This is consistent with the notion derived from experimental work that myocardial cell proliferation and coronary vascular growth rely on distinct developmental programs. Our observations, together with data in the literature on elasmobranchs, support the view that the mixed ventricular myocardium is primitive for chondrichthyans. The reduction or even lack of compacta in holocephali has to be regarded as a derived anatomical trait. Our findings also fit in with the view that the mixed myocardium was the primitive condition in gnathostomes, and that the absence of compact ventricular myocardium in different actinopterygian groups is the result of a repeated loss of such type of cardiac muscle during fish evolution.