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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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Previous work on the anatomy of embryonic and adult hearts of the shark Galeus atlanticus showed that the cardiac outflow tract consists not of a single component but of two, the myocardial conus arteriosus and the nonmyocardial bulbus arteriosus. However, it was still uncertain whether a bulbus occurs regularly in elasmobranchs. The present study was designed to gain new insight into this issue. Moreover, we investigated the interspecific variation in the relative length of the bulbus in order to decide whether such variation is related to the lifestyle or to the phylogenetic position of sharks and rays. The material examined consisted of hearts of 42 species belonging to seven families of squalomorphs, six families of galeomorphs and six families of batoids. Our findings confirmed that the bulbus arteriosus is indeed a cardiac outflow tract component commonly present in all elasmobranchs examined. In addition, we propose that the relative length of the bulbus arteriosus is rather related to the phylogenetic position of the species than to its lifestyle. Overall, the bulbus arteriosus is notably longer in galeomorphs than in squalomorphs and batoids, with the exception of the two representatives of Torpedo , which possess a bulbus arteriosus longer than other batoids. Likewise, the anatomical origin of the outflow tract from the ventricle is closely related to the relative length of the bulbus. We suggest that both anatomical traits are associated with the phylogenetic position of the sharks and rays investigated. [ABSTRACT FROM AUTHOR]

A cell line has been developed from the bulbus arteriosus (BA) of the walleye (WE), Sander vitreus (Mitchill), and is termed WEBA. WEBA produced collagen I, and when held at confluency for days or weeks, spontaneously formed capillary-like tubes. WEBA cells bound fluorescently-labeled Ulex europaeus lectin agglutinin I (UEA-1), took up acetylated low density lipoprotein (Ac-LDL), were stained for von Willebrand factor (vWF), and produced nitric oxide (NO). The cytoskeleton consisted at least of α- and β-tubulin, vimentin, and actin, with the actin organized into circumferential bundles. Immunofluorescence staining revealed at least two tight junction proteins, zonula occludens-1 (ZO-1) and claudin 3. Together these results suggest that WEBA is an endothelial cell line. Relatively high doses of 2,3,7,8-tetrachlorodibenzodioxin (TCDD) induced cytochrome P4501A (CYP1A) protein and 7-ethoxyresorufin o-deethylase (EROD) activity in WEBA. As one of the first fish endothelial and BA cell lines, WEBA should be useful in many disciplines in which the teleost cardiovascular system is a focus. [ABSTRACT FROM AUTHOR]

It has been reported that in chondrichthyans the cardiac outflow tract is composed of the myocardial conus arteriosus, while in most teleosteans it consists of the nonmyocardial bulbus arteriosus. Classical studies already indicated that a conus and a bulbus coexist in several ancient actinopterygian and teleost groups. Recent work has shown that a cardiac outflow tract consisting of a conus and a bulbus is common to both cartilaginous and bony fishes. Nonetheless and despite their position at the base of the actinopterygian phylogenetic lineage, the anatomical arrangement of the cardiac outflow tract of the Polypteriformes remained uncertain. The present study of hearts from gray bichirs was intended to fill this gap. The cardiac outflow tract of the bichir consists of two main components, namely a very long conus arteriosus, furnished with valves, and a short, intrapericardial, arterial-like bulbus arteriosus, which differs from the ventral aorta because it is covered by epicardium, shows a slightly different spatial arrangement of the histological elements and is crossed by coronary arteries. Histomorphologically, the outflow tract consists of three longitudinal regions, distal, middle and proximal, an arrangement which has been suggested to be common to all vertebrates. The distal region corresponds to the bulbus, while the conus comprises the middle and proximal regions. The present findings reinforce the notion that the bulbus arteriosus of fish has played an essential role in vertebrate heart evolution as it is the precursor of the intrapericardial trunks of the aorta and pulmonary artery of birds and mammals. [ABSTRACT FROM AUTHOR]

Abstract: Recent work has shown that the outflow tract (OFT) of the chondrichthyan heart does not consist of a single component, the conus arteriosus, as has been classically accepted, but two, the myocardial conus arteriosus and the non-myocardial bulbus arteriosus. The aim of this study was to describe the embryonic development of the bulbus, under the assumption that the heart of extant chondrichthyans represents the primitive condition of this organ in jawed vertebrates. The study was carried out in Atlantic sawtail catshark embryos belonging to developmental stages 20–34 of Ballard et al. (1993). They were examined using histochemical and immunohistochemical techniques for light and fluorescence microscopy. The entire OFT displays a myocardial character during early development. At stage 28, a new, non-myocardial segment forms at the distal end of the OFT. This segment, which is the precursor of the bulbus arteriosus, shows positive reactivity for 4,5-diaminofluorescein 2-diacetate throughout development from stage 31. Smooth muscle α-actin-positive cells appear in the prospective bulbus at stage 33; they markedly increase in number during stage 34. Subsequently, they extend backwards to overlap with the conal myocardium, giving rise to a myocardium and smooth muscle interface that has been described elsewhere as the middle component of the vertebrate OFT. Our embryological findings in a elasmobranch species are consistent with the notion that a second heart field has existed since the beginning of the jawed vertebrate radiation and that the bulbus arteriosus of chondrichthyans is a second heart field-derived structure. [Copyright &y& Elsevier]

Abstract: This report analyzes the localization of collagen and elastin in the teleost bulbus arteriosus by histochemistry and by transmission electron microscopy. Martin''s trichrome staining shows widespread distribution of collagen in the wall of the bulbus. However, Sirius red indicates that collagen is mostly restricted to the valves and to the subepicardial layer. This is confirmed by transmission electron microscopy. Trichrome staining gives false positives that may be related to the chemical characteristics of both matrix components and dyes. By contrast, Sirius red constitutes a highly reliable method to detect collagen distribution. On the other hand, orcein heavily stains the bulbus of all teleosts examined. This includes the bulbus of the Antarctic teleosts, which do not show structurally discernable elastin fibers. In these cases, orcein may be staining non-elastin components, or basic elastin components not assembled into larger units. In the teleost bulbus, accurate identification of collagen and elastin cannot be based solely on histochemistry, but should be accompanied by structural identification of the components under study. [Copyright &y& Elsevier]

Abstract: Immunostimulating effects of water extract from bulbus arteriosus in tuna were examined and found to enhance IgM production by human hybridoma HB4C5 cells. When tuna bulbus arteriosus extract was heated, the IgM production-stimulating activity was decreased or lost. To identify the active substance, the extract was partially purified by anion-exchange chromatography, and proteins contained in positive fractions were analyzed by LC-MS/MS. The positive fractions contained lactate dehydrogenase, triosephosphate isomerase, enolase, and haemoglobin. Each of these four proteins was verified to enhance IgM production by HB4C5 cells with the commercially available purified proteins, suggesting that all of these four proteins are active substances in the extract. The immunostimulatory effect of these proteins was also examined on mouse primary spleen lymphocytes in vitro. Lactate dehydrogenase enhanced IgA, IgG, and IgM production and triosephosphate isomerase enhanced IgA and IgG production. In particular, lactate dehydrogenase had the strongest Ig production-stimulating effect on HB4C5 cells and on mouse primary lymphocytes. Thus, fish viscera may serve as an important raw material for the enhancement of the acquired immune system, once processed. [Copyright &y& Elsevier]

Abstract: Elastin is the extracellular matrix protein responsible for properties of extensibility and elastic recoil in large blood vessels, lung and skin of most vertebrates. Elastin is synthesized as a monomer, tropoelastin, but is rapidly transformed into its final polymeric form in the extracellular matrix. Until recently information on sequence and developmental expression of tropoelastins was limited to mammalian and avian species. We have recently identified and characterized two expressed tropoelastin genes in zebrafish. This was the first example of a species with multiple tropoelastin genes, raising the possibility of differential expression and function of these tropoelastins in elastic tissues of the zebrafish. Here we have investigated the temporal expression and tissue distribution of the two tropoelastin genes in developing and adult zebrafish. Expression was detected early in skeletal cartilage structures of the head, in the developing outflow tract of the heart, including the bulbus arteriosus and the ventral aorta, and in the wall of the swim bladder. While the temporal pattern of expression was similar for both genes, the upregulation of eln2 was much stronger than that of eln1. In general, both genes were expressed and their gene products deposited in most of the elastic tissues examined, with the notable exception of the bulbus arteriosus in which eln2 expression and its gene product was predominant. This finding may represent a sub-specialization of eln2 to provide the unique architecture of elastin and the specific mechanical properties required by this organ. [Copyright &y& Elsevier]