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Cardiac abnormalities may pose a threat to salmonid aquaculture due to their potential detrimental effect on fish health and welfare. The teleost heart is an extremely plastic organ with important morphological differences between wild and farmed fish that include ventricular shape, alignment of the bulbus arteriosus and epicardial fat deposition. However, little is known about how different factors and interactions among them may affect cardiac morphology of Atlantic salmon. To determine whether rearing temperature could induce cardiac malformations in large Tasmanian Atlantic salmon, we examined a range of cardiac morphology indicators and growth parameters in a population of 1-2 kg seawater salmon (n = 60 temperature-1 diet-1 ) exposed to control and elevated temperatures of 15 and 19°C, respectively, while fed one of two commercial feeds with different dietary energy levels. Most fish possessed conspicuous fat around the heart with a tendency towards a rounded ventricle and a more obtuse angle of the bulbus arteriosus. However, fish showed no significant differences in heart shape and bulbus alignment in relation to water temperature and dietary energy. These results suggest that cardiac morphology of large Atlantic salmon is unlikely to be affected by rearing temperature and dietary energy during the grow-out phase.

Adrenergic regulation, acting via the sympathetic nervous system, provides a major mechanism to control cardiac function. It has recently been shown that hypoxia inducible factor-1α (Hif-1α) is necessary for normal development of sympathetic innervation and control of cardiac function in the mouse. To investigate whether this may represent a fundamental trait shared across vertebrates, we assessed adrenergic regulation of the heart in wild-type and Hif-1α knockout (hif-1α −/−) zebrafish (Danio rerio). Wild-type and hif-1α −/− zebrafish larvae (aged 4 and 7 days postfertilisation) exhibited similar routine heart rates within a given age group, and β-adrenergic receptor blockade with propranolol universally reduced heart rate to comparable levels, indicating similar adrenergic tone in both genotypes. In adult fish, in vivo heart rate measured during anaesthesia was identical between genotypes. Treatment of spontaneously beating hearts in vitro with adrenaline revealed a similar positive chronotropic effect and similar maximum heart rates in both genotypes. Tyrosine hydroxylase immunohistochemistry with confocal microscopy demonstrated that the bulbus arteriosus (outflow tract of the teleost heart) of adult fish was particularly well innervated by sympathetic nerves, and nerve density (as a percentage of bulbus arteriosus area) was similar between wild-types and hif-1α −/− mutants. In summary, we did not find any evidence that adrenergic cardiac control was perturbed in larval or adult zebrafish lacking Hif-1α. We conclude that Hif-1α is not essential for the normal development of cardiovascular control or adult sympathetic cardiac innervation in zebrafish, although it is possible that it plays a redundant or auxiliary role.

A new, three-dimensional geometric morphometric approach was assessed to study the effect of developmental temperature on fish heart shape utilizing geometric morphometrics of three-dimensional landmarks captured on digitally reconstructed zebrafish hearts. This study reports the first three-dimensional analysis of the fish heart and demonstrates significant shape modifications occurring after three developmental temperature treatments (TD = 24, 28 or 32°C) at two distinct developmental stages (juvenile and adult fish). Elevation of TD induced ventricle roundness in juveniles, males and females. Furthermore, significant differences that have not been described so far in heart morphometric indices (i.e., ventricle sphericity, bulbus arteriosus elongation and relative location, heart asymmetry) were identified. Sex proved to be a significant regulating factor of heart shape plasticity in response to TD. This methodology offers unique benefits by providing a more precise representation of heart shape changes in response to developmental temperature that are otherwise not discernable with the previously described two-dimensional methods. Our work provides the first evidence of three-dimensional shape alterations of the zebrafish heart adding to the emerging rationale of temperature-driven plastic responses of global warming and seasonal temperature disturbances in wild fish populations and in other ectothermic vertebrates as well (amphibians and reptiles).

To protect the gill capillaries from high systolic pulse pressure, the fish heart contains a compliant non-contractile chamber called the bulbus arteriosus which is part of the outflow tract (OFT) which extends from the ventricle to the ventral aorta. Thermal acclimation alters the form and function of the fish atria and ventricle to ensure appropriate cardiac output at different temperatures, but its impact on the OFT is unknown. Here we used ex vivo pressure–volume curves to demonstrate remodelling of passive stiffness in the rainbow trout ( Oncorhynchus mykiss ) bulbus arteriosus following more than eight weeks of thermal acclimation to 5, 10 and 18°C. We then combined novel, non-biased Fourier transform infrared spectroscopy with classic histological staining to show that changes in compliance were achieved by changes in tissue collagen-to-elastin ratio. In situ gelatin zymography and SDS-PAGE zymography revealed that collagen remodelling was underpinned, at least in part, by changes in activity and abundance of collagen degrading matrix metalloproteinases. Collectively, we provide the first indication of bulbus arteriosus thermal remodelling in a fish and suggest this remodelling ensures optimal blood flow and blood pressure in the OFT during temperature change.

Background Cardiomyopathy is a kind of cardiovascular diseases, which makes it more difficult for the heart to pump blood to other parts of the body, eventually leading to heart failure. Naoxintong (NXT), as a traditional Chinese Medicine (TCM) preparation, is widely used in the treatment of cardiovascular diseases, including cardiomyopathy, while its underlying mechanism has not been fully elucidated. The purpose of this study is to investigate the therapeutic effect of NXT on cardiomyopathy and its molecular mechanism in zebrafish model. Methods The zebrafish cardiomyopathy model was established using terfenadine (TFD) and treated with NXT. The therapeutic effect of NXT on cardiomyopathy was evaluated by measuring the heart rate, the distance between the sinus venosus and bulbus arteriosus (SV-BA), the pericardial area, and the blood flow velocity of zebrafish. Then, the zebrafish hearts were isolated and collected; transcriptome analysis of NXT on cardiomyopathy was investigated. Moreover, the heg1 mutant of zebrafish congenital cardiomyopathy model was used to further validate the therapeutic effect of NXT on cardiomyopathy. Additionally, UPLC analysis combined with the zebrafish model investigation was performed to identify the bioactive components of NXT. Results In the TFD-induced zebrafish cardiomyopathy model, NXT treatment could significantly restore the cardiovascular malformations caused by cardiac dysfunction. Transcriptome and bioinformatics analyses of the TFD and TFD + NXT treated zebrafish developing hearts revealed that the differentially expressed genes were highly enriched in biological processes such as cardiac muscle contraction and heart development. As a cardiac development protein associated with cardiomyopathy, HEG1 had been identified as one of the important targets of NXT in the treatment of cardiomyopathy. The cardiovascular abnormalities of zebrafish heg1 mutant could be recovered significantly from NXT treatment, including the expanded atrial cavity and blood stagnation. qRT-PCR analysis further showed that NXT could restore cardiomyopathy phenotype in zebrafish through HEG1-CCM signaling. Among the seven components identified in NXT, paeoniflorin (PF) and salvianolic acid B (Sal B) were considered to be the main bioactive ones with myocardial protection. Conclusion NXT presented myocardial protective effect and could restore myocardial injury and cardiac dysfunction in zebrafish; the action mechanism was involved in HEG1-CCM signaling.

Made available in DSpace on 2019-10-06T16:43:05Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-01-01 The cardiac outflow tract (OFT) in teleosts is composed of a proximal short conus arteriosus and a distal well-developed bulbus arteriosus located between the ventricle and ventral aorta. The role of these anatomical components includes structural connections, prevention of blood backflow and blood pressure control, which are related to their histological and histochemical compositions. A previous study in the heart of the Amazonian species Arapaima gigas reported an unusual OFT arrangement among teleosts that has been found only in members of Osteoglossiformes so far. Thus, considering the wide structural variability of the teleostean OFT, the present study focuses on identifying glycosaminoglycans types and describing the distribution of collagen, elastic and reticular fibers in the conus arteriosus, conal valves, and bulbus arteriosus of A. gigas. Hearts from A. gigas between 327 and 4040 g weight were used. Collagen fibers were concentrated in regions that were regularly exposed to stress, as elastic fibers showed a broad distribution in all anatomical segments. Several fibrous connections between the conal valve leaflets and the conus arteriosus were observed, possibly acting as the primary connection form. The functions of the connective fibers in the valve leaflets and the bulbus are supported by an extracellular matrix rich in non-sulfated glycosaminoglycans. The complex reticular fiber network in the compact myocardium of the conus and the smooth muscle of the bulbus wall suggests a relevant role in support contraction of both muscle types, and in attachment and mobility of the conal valve leaflets. Department of Zoology Institute of Biosciences São Paulo State University (Unesp) Department of Biological Sciences School of Sciences São Paulo State University (Unesp) Aquaculture Center of Unesp São Paulo State University (Unesp) Tocantins Federal University (UFT) Araguaína Campus Department of Biology Institute of Biosciences Humanities and Exact Sciences São Paulo State University (Unesp) Department of Zoology Institute of Biosciences São Paulo State University (Unesp) Department of Biological Sciences School of Sciences São Paulo State University (Unesp) Aquaculture Center of Unesp São Paulo State University (Unesp) Department of Biology Institute of Biosciences Humanities and Exact Sciences São Paulo State University (Unesp)

Polypyrrole-based polyamides are used as sequence-specific DNA probes. However, their cellular uptake and distribution are affected by several factors and have not been extensively studied in vivo . Here, we generated a series of fluorescence-conjugated polypyrrole compounds and examined their cellular distribution using live zebrafish and cultured human cells. Among the evaluated compounds, Py 3 -FITC was able to visualize collagen-rich tissues, such as the jaw cartilage, opercle and bulbus arteriosus, in early-stage living zebrafish embryos. Then, we stained cultured human cells with Py 3 -FITC and found that the staining became more intense as the amount of collagen was increased. In addition, Py 3 -FITC-stained HR cells, which represent a type of ionocyte on the body surface of living zebrafish embryos. Py 3 -FITC has low toxicity, and collagen-rich tissues and ionocytes can be visualized when soaked in Py 3 -FITC solution. Therefore, Py 3 -FITC may be a useful live imaging tool for detecting changes in collagen-rich tissue and ionocytes, including their mammalian analogues, during both normal development and disease progression.

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.

Detection of patterns of subcellular calcium distribution in the cardiovascular system can contribute to understanding its role in cardiac and blood function. The present study localized calcium in heart atrium, ventricle, and bulbus arteriosus as well as in erythrocytes of zebrafish Danio rerio using an oxalate-pyroantimonate technique combined with transmission electron microscopy. Intracellular calcium stores were detected in caveolae, mitochondria, and the nuclei of several zebrafish cardiac cell types. Melanin pigmentation containing calcium stores was detected in the pericardial cavity. Melanin might be an extracellular source of calcium for heart beating and/or a lubricant to prevent friction during beating process. Calcium deposits were also detected in the plasma membrane, cytoplasm and nucleus of erythrocytes as well as in blood plasma. Possible exchange of calcium between erythrocytes and blood plasma was observed. Interactions of such calcium stores and possible contribution of extracellular calcium stores such as melanin pigmentation to supply calcium for vital functions of heart cells should be addressed in future studies.

The ventricle of the fish heart is a chamber that exhibits great morphological and vascular variability among species. However, although the Neotropical region has the greatest taxonomic and functional diversity in freshwater fish, many considerations have been formed without previous knowledge of the ventricular morphology of these fishes. Thus, the purpose of the present study was to describe the anatomy, myoarchitecture, and distribution of coronary vessels in the ventricle of three species belonging to two representative groups from this geographical area, Leporinus elongatus, Hoplias malabaricus (Characiformes) and Pterodoras granulosus (Siluriformes), using gross anatomy and light microscopy. The species L. elongatus and H. malabaricus presented a pyramidal ventricle associated to a mixed myocardium, formed by compact and spongy layers. The mixed myocardium was also observed in P. granulosus, but associated with a sac-like ventricle. The compact layer of the species studied was formed by muscular bundles in longitudinal and circular disposition. The spongy layer constituted most of the ventricular myocardium and was formed by a complex network of trabecular sheets presenting muscle fibers also in longitudinal and circular disposition. Coronary vessels were present in the three species and were observed primarily on the surface of the bulbus arteriosus, later branching on the ventricular surface and penetrating the myocardium only at the compact layer level. These characteristics allow classification of the ventricles studied as type II. Although the type I ventricle is the most common type in teleosts, it is important to emphasize that this type has not been observed in any Neotropical freshwater teleosts studied to date.

Prothioconazole is a fungicide that has been widely used in general agriculture and livestock husbandry. This study evaluated the acute toxicity of prothioconazole to zebrafish embryos by assessing their hatching rate and malformation when exposed to different concentrations of prothioconazole. The 96 h-LC50 value of zebrafish embryos was 1.70 mg/L. Upon exposure to 0.85 mg/L, the mortality rate of the embryos significantly increased while their hatching rate decreased significantly. At prothioconazole concentrations higher than 0.43 mg/L, developmental morphologic abnormalities such as heart and yolk-sac edema, spine curvature, tail deformity, shortened body length and decreased eye area were observed. The heart rate of embryos decreased in a dose-dependent fashion during the exposure time. Prothioconazole exposure also resulted in increased rates of cardiac malformation detected by significant increase in the distance between the sinus venosus and bulbus arteriosus and the pericardium area. Moreover, the expression levels of genes related to cardiac development (amhc, vmhc, fli1, hand2, gata4, nkx2.5, tbx5 and atp2a2a) were significantly altered after exposure to prothioconazole. Indeed, this study revealed the adverse effects on the developmental and cardiovascular system of zebrafish embryo caused by prothioconazole. It further elucidated the risk of prothioconazole exposure to vertebrate cardiovascular toxicity. As such, it provides a theoretical foundation for pesticide risk management measures.

Mechanical forces are well known for modulating heart valve developmental programs. Yet, it is still unclear how genetic programs and mechanosensation interact during heart valve development. Here, we assessed the mechanosensitive pathways involved during zebrafish outflow tract (OFT) valve development in vivo. Our results show that the hippo effector Yap1, Klf2, and the Notch signaling pathway are all essential for OFT valve morphogenesis in response to mechanical forces, albeit active in different cell layers. Furthermore, we show that Piezo and TRP mechanosensitive channels are important factors modulating these pathways. In addition, live reporters reveal that Piezo controls Klf2 and Notch activity in the endothelium and Yap1 localization in the smooth muscle progenitors to coordinate OFT valve morphogenesis. Together, this work identifies a unique morphogenetic program during OFT valve formation and places Piezo as a central modulator of the cell response to forces in this process. journal article research support, non-u.s. gov't 2019 09 16 2019 09 16 imported

Summary: The hormone αKlotho regulates lifespan in mice, as knockouts die early of what appears to be accelerated aging due to hyperphosphatemia and soft tissue calcification. In contrast, the overexpression of αKlotho increases lifespan. Given the severe mouse phenotype, we generated zebrafish mutants for αklotho as well as its binding partner fibroblast growth factor-23 (fgf23). Both mutations cause shortened lifespan in zebrafish, with abrupt onset of behavioral and degenerative physical changes at around 5 months of age. There is a calcification of vessels throughout the body, most dramatically in the outflow tract of the heart, the bulbus arteriosus (BA). This calcification is associated with an ectopic activation of osteoclast differentiation pathways. These findings suggest that the gradual loss of αKlotho found in normal aging might give rise to ectopic calcification. : αKlotho regulates mineral homeostasis and affects lifespans in mammals. Singh et al. show that a loss of αklotho in zebrafish results in reduced lifespans and vascular calcification in the outflow tract of the heart. Vascular calcification is associated with an upregulation of bone remodeling pathways and osteoclast differentiation. Keywords: αKlotho, Klotho, FGF23, aging, calcification, cardiovascular system, zebrafish

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.

Particulate matter (PM10) is one of the most important indicators of the pollution that characterizes air quality. Epidemiological studies have shown that PM10 can cause cardiovascular-related diseases in the population. And, we studied the developmental toxicity of PM10 and the underlying mechanism of its effects on the cardiovascular system of zebrafish embryo/larva. Changes in cardiac morphology, sinus venosus and bulbus arteriosus (SV-BA) distance, heart rate, vascular subintestinalis, blood flow, returned blood volume, and reactive oxygen species (ROS) level were measured, and changes in the expression levels of certain genes were assessed via RT-PCR. The results showed that PM10 caused a significant increase in pericardial sac area and SV-BA distance, a decrease in heart rate, inhibition of vascular subintestinalis growth, blood flow obstruction, reduced venous return, and other cardiovascular toxicities. PM10 induced an increase in the ROS level and significant increases in the expression levels of ERS signalling pathway factors and Nrf2 signalling pathway factors. The expression levels of the Wnt pathway-related genes also showed significant changes. Furthermore, ROS inhibitor N-Acetyl-l-cysteine (NAC) could ameliorate the cardiovascular toxicity of PM10 in zebrafish larvae. It is speculated that PM10 may result in cardiovascular toxicity by inducing higher ROS levels in the body, which could then induce ERS and lead to defects in the expression of genes related to the Wnt signalling pathway. The Nrf2 signalling pathway was activated as a stress compensatory mechanism during the early stage of PM10-induced cardiovascular injury. However, it was insufficient to counteract the PM10-induced cardiovascular toxicity.

Metalaxyl is an anilide pesticide that is widely used to control plant diseases caused by Peronosporales species. In order to study the toxic effects, zebrafish embryos were exposed to metalaxyl at nominal concentrations of 5, 50 and 500 ng/L for 72 hr, and the cardiac development and functioning of larvae were observed. The results showed that metalaxyl exposure resulted in increased rates of pericardial edema, heart hemorrhage and cardiac malformation. The distance between the sinus venosus and bulbus arteriosus, stroke volume, cardiac output and heart rate were significantly increased in larvae exposed to 50 and 500 ng/L metalaxyl compared to solvent control larvae. Significant upregulation in the transcription of tbx5, gata4 and myh6 was observed in the 50 and 500 ng/L treatments, and that of nkx2.5 and myl7 was observed in the 5, 50 and 500 ng/L groups. These disturbances may be related to cardiac developmental and functional defects in the larvae. The activity of Na+/K+-ATPase and Ca2+-ATPase was significantly increased in zebrafish embryos exposed to 500 ng/L metalaxyl, and the mRNA levels of genes related to ATPase (atp2a11, atp1b2b, and atp1a3b) (in the 50 and 500 ng/L groups) and calcium channels (cacna1ab) (in the 500 ng/L group) were significantly downregulated; these changes might be associated with heart arrhythmia and functional failure.

© 2015 Elsevier Inc.. Cardiac neural crest cells are essential for outflow tract remodeling in animals with divided systemic and pulmonary circulatory systems, but their contributions to cardiac development in animals with a single-loop circulatory system are less clear. Here we genetically labeled neural crest cells and examined their contribution to the developing zebrafish heart. We identified two populations of neural crest cells that contribute to distinct compartments of zebrafish cardiovascular system at different developmental stages. A stream of neural crest cells migrating through pharyngeal arches 1 and 2 integrates into the myocardium of the primitive heart tube between 24 and 30. h post fertilization and gives rise to cardiomyocytes. A second wave of neural crest cells migrating along aortic arch 6 envelops the endothelium of the ventral aorta and invades the bulbus arteriosus after three days of development. Interestingly, while inhibition of FGF signaling has no effect on the integration of neural crest cells to the primitive heart tube, it prevents these cells from contributing to the outflow tract, demonstrating disparate responses of neural crest cells to FGF signaling. Furthermore, neural crest ablation in zebrafish leads to multiple cardiac defects, including reduced heart rate, defective myocardial maturation and a failure to recruit progenitor cells from the second heart field. These findings add to our understanding of the contribution of neural crest cells to the developing heart and provide insights into the requirement for these cells in cardiac maturation.

In the vertebrate heart the intracardiac nervous system is the final common pathway for autonomic control of cardiac output, but the neuroanatomy of this system is not well understood. In this study we investigated the innervation of the heart in a model vertebrate, the zebrafish. We used antibodies against acetylated tubulin, human neuronal protein C/D, choline acetyltransferase, tyrosine hydroxylase, neuronal nitric oxide synthase, and vasoactive intestinal polypeptide to visualize neural elements and their neurotransmitter content. Most neurons were located at the venous pole in a plexus around the sinoatrial valve; mean total number of cells was 197 ± 23, and 92% were choline acetyltransferase positive, implying a cholinergic role. The plexus contained cholinergic, adrenergic, and nitrergic axons and vasoactive intestinal polypeptide-positive terminals, some innervating somata. Putative pacemaker cells near the plexus showed immunoreactivity for hyperpolarization-activated cyclic nucleotide-gated channel 4 (HCN4) and the transcription factor Islet-1 (Isl1). The neurotracer neurobiotin showed that extrinsic axons from the left and right vagosympathetic trunks innervated the sinoatrial plexus proximal to their entry into the heart; some extrinsic axons from each trunk also projected into the medial dorsal plexus region. Extrinsic axons also innervated the atrial and ventricular walls. An extracardiac nerve trunk innervated the bulbus arteriosus and entered the arterial pole of the heart to innervate the proximal ventricle. We have shown that the intracardiac nervous system in the zebrafish is anatomically and neurochemically complex, providing a substrate for autonomic control of cardiac effectors in all chambers. J. Comp. Neurol. 523:1683–1700, 2015. © 2015 Wiley Periodicals, Inc.

Aims To determine the prophylactic effects of an elastin peptide derived from the bulbus arteriosus of bonitos and prolylglycine (PG), a degradation product of elastin peptide, on vascular dysfunction in spontaneously hypertensive rats (SHRs). Main methods Male 15-week-old SHR/Izm rats were fed without (control group) or with elastin peptide (1 g/kg body weight) for 5 weeks (EP group), or were infused via an osmotic mini-pump for 4 weeks with PG (PG group) or saline (control group). Using thoracic aortas, we assessed endothelial changes by scanning electron microscopy. Vascular reactivity (contraction and relaxation) and pressure-induced distension was compared. mRNA production levels of endothelial nitric oxide synthase (eNOS) and intercellular adhesion molecule-1 (ICAM-1) were investigated by real-time-polymerase chain reaction. Key findings Aortas of the EP group displayed limited endothelial damage compared with that in the control group. Under treatment of SHRs with elastin peptide, the effect of phenylephrine returned closer to the normal level observed in normotensive Wistar–Kyoto (WKY/Izm) rats. mRNA production of eNOS (but not ICAM-1) was greater in the EP group than in the control group. Endothelial damage was suppressed and pressure-induced vascular distension was greater in the PG group than in the corresponding control group. Significance These results suggest that elastin peptide from bonitos elicits prophylactic affects hypertension-associated vascular dysfunction by targeting the eNOS signaling pathway. PG may be a key mediator of the beneficial effects of elastin peptide.

The vertebrate heart develops from several progenitor lineages. After early-differentiating first heart field (FHF) progenitors form the linear heart tube, late-differentiating second heart field (SHF) progenitors extend the atrium and ventricle, and form inflow and outflow tracts (IFT/OFT). However, the position and migration of late-differentiating progenitors during heart formation remains unclear. Here, we track zebrafish heart development using transgenics based on the cardiopharyngeal gene tbx1. Live imaging uncovers a tbx1 reporter-expressing cell sheath that continuously disseminates from the lateral plate mesoderm towards the forming heart tube. High-speed imaging and optogenetic lineage tracing corroborates that the zebrafish ventricle forms through continuous addition from the undifferentiated progenitor sheath followed by late-phase accrual of the bulbus arteriosus (BA). FGF inhibition during sheath migration reduces ventricle size and abolishes BA formation, refining the window of FGF action during OFT formation. Our findings consolidate previous end-point analyses and establish zebrafish ventricle formation as a continuous process., Late-differentiating second heart field progenitors contribute to atrium, ventricle, and outflow tract in the zebrafish heart but how remains unclear. Here, the authors image heart formation in transgenics based on the cardiopharyngeal gene tbx1 and show that progenitors are continuously added.

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.

The bulbus arteriosus is a compliant structure between the ventricle and ventral aorta of teleost fish. It serves as a “wind-kessel” that dampens pressure variations during the cardiac cycle allowing a continuous flow of blood into the gills. The bulbus arteriosus receives sympathetic innervation and is affected by several circulating substances, indicating neurohumoral control. We have previously shown that the peptide hormone, cholecystokinin (CCK), affects the hemodynamics of the cardiovascular system in rainbow trout ( Oncorhynchus mykiss ) by increasing flow pulse amplitude without affecting cardiac output. We hypothesized that this could be explained by an altered tonus or compliance/distensibility of the bulbus arteriosus. Our results show that there is a substantial effect of CCK on the bulbus arteriosus. Concentrations of CCK that altered the cardiac function of in situ perfused hearts also contracted the bulbus arteriosus in vitro. Pressure–volume curves revealed a change in both the tonus and the compliance/distensibility of this structure. Furthermore, the stimulatory (constricting) effect of CCK was also evident in the ventricle and vasculature leading to the gills, but absent in the atrium, efferent branchial arteries and dorsal aorta. In conclusion, CCK alters the mechanical properties of the ventricle, bulbus arteriosus, ventral aorta and afferent gill vasculature, thus maintaining adequate branchial and systemic blood flow and pressure when cardiorespiratory demands change, such as after feeding.

For several species of fish myxosporeans known at present, the musculature has been designated as the location of intrapiscine development. In the majority of these cases, plas- modia and spores are actually found in the muscle cells, but there are also myxosporeans that select intermuscular connective tissue, fish bones, nerves and capillaries of the musculature as their site of development. During the plasmodial development of Myxobolus, Henneguya and Thelohanellus species in fish inhabiting Hungarian freshwaters, 3 main locations of development inside the muscles were identified. Pseudodispar-type plasmodia, such as M. cyprini, M. musculi and M. pseudodispar , form plasmodia intracellularly in the muscle cells, while the plasmodia of M. pfeifferi, M. sandrae and T. hovorkai develop in the intermuscular connective tissue. A similar development in the connective tissue of the ventricle and the bulbus arteriosus was observed for M. dogieli, a heart parasite found in some cyprinid fishes. The third type of development is repre- sented by M. tauricus, which prefers the collagenous elements of the fin rays, but its plasmodia are commonly found in the muscle attached to the fish bones.

Heart morphology is particularly plastic in teleosts and differs between farmed and wild Atlantic salmon. However, little is known about how different culture practices and sex affect heart morphology. This study investigated how vaccination, triploidy and sex affected heart size and heart morphology (ventricle shape, angle of the bulbus arteriosus) in farmed Atlantic salmon for 18 months following vaccination (from c. 50-3000 g body weight). In addition, hearts were examined histologically after 7 months in sea water. All fish sampled were sexually immature. Vaccinated fish had significantly heavier hearts relative to body weight and a more triangular ventricle than unvaccinated fish, suggesting a greater cardiac workload. Irrespective of time, triploids had significantly heavier hearts relative to body weight, a more acute angle of the bulbus arteriosus and less fat deposition in the epicardium than diploids. The ventricle was also more triangular in triploids than diploids at seawater transfer. Sex had transient effects on the angle of the bulbus arteriosus, but no effect on relative heart weight or ventricle shape. From a morphological perspective, the results indicate that vaccination and triploidy increase cardiac workload in farmed Atlantic salmon.

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.

This chapter covers morphological aspects of the fish heart. It starts with the recognition of the existence of six heart components that are present during embryonic development, and in the adult, and that are anatomically arranged in sequence: sinus venosus, atrium, atrioventricular canal, ventricle, conus arteriosus, and bulbus arteriosus. The anatomy and structure of all the cardiac components are then described and compared between the different fish groups. Data from the basal Cyclostomata, chondrichthyans, basal Osteichthyes, and more advanced teleosts are included and analyzed. Then, data on blood supply to the heart, nerves, and localization of the heart pacemaker are discussed. A separate section at the end of the chapter is committed to the lungfish heart. Lungfishes share many anatomical and functional characteristics with both freshwater fish and amphibians, and the lungfish heart shows some unique and very peculiar characteristics. The enormous diversification of morphological characteristics of the fish heart is emphasized throughout this chapter.

Francisella noatunensis subsp. orientalis (Fno) (syn. F. asiatica) is an emergent warmwater fish pathogen and the causative agent of piscine francisellosis. Although Fno causes septicemia and can live extracellularly in infected tilapia (Oreochromis spp.), the early interaction of Fno with vasculature endothelium is unknown. In the present study, we examined the interaction of wild-type Fno (WT) and two Fno knockout [intracellular growth loci C (ΔiglC) and pathogenicity determinant protein A (ΔpdpA)] strains with the endothelial O. mossambicus bulbus arteriosus cell line (TmB) at 25 °C and 30 °C. Similar amounts of WT, ΔiglC, and ΔpdpA attached and were detected intracellularly after 5 h of incubation at both temperatures; however temperature affected attachment and uptake. While significantly greater amounts of Fno (WT, ΔiglC, and ΔpdpA) were detected intracellularly when TmB cells were incubated at 30 °C, bacteria attached to TmBs at greater levels at 25 °C. Only WT Fno was able to replicate intracellularly at 25 °C, which resulted in Fno mediated cytotoxicity and apoptosis at 24 and 72 h post-infection. WT Fno incubated at 30 °C as well as ΔiglC, and ΔpdpA incubated at 25 °C and 30 °C were all defective for survival, replication, and the ability to cause cytotoxicity in TmB. Taken together, these results demonstrate that temperature plays a vital role for Fno intracellular survival, persistence and cytotoxicity.