The lungs of Polypterus senegalus and Erpetoichthys calabaricus: Insights into the structure and functional distribution of the pulmonary epithelial cells.

The present article is a comparative, structural study of the lung of Polypterus senegalus and Erpetoichthys calabaricus, two species representative of the two genera that constitute the Polypteriformes. The lung of the two species is an asymmetric, bi-lobed organ that arises from a slit-like opening in the ventral side of the pharynx. The wall is organized into layers, being thicker in P. senegalus. The inner epithelium contains ciliated and non-ciliated bands. The latter constitute the respiratory surface and are wider in E. calabaricus. The air-blood barrier is thin and uniform in P. senegalus and thicker and irregular in E. calabaricus. In the two species, the ciliated areas contain ciliated cells, mucous cells and cells with lamellar bodies. Additionally, P. senegalus contains polymorphous granular cells (PGCs) and neuroendocrine cells (NECs) while E. calabaricus lacks PGCs but shows granular leukocytes and a different type of NEC. Interestingly, ciliated cells and secretory cells show a dual morphology in E. calabaricus indicating the presence of cellular subtypes and suggesting more complex secretory activity. Also in E. calabaricus, cilia show a novel doublet-membrane interaction that may control the displacement of the microtubule doublets. The subepithelium is a connective layer that appears thicker in P. senegalus and contains, in the two species, fibroblasts and granulocytes. The outer layer contains bundles of richly innervated striated muscle. This layer is likely involved in the control of lung motion. In the two species, smooth muscle cells constitute a limiting layer between the subepithelium and the striated muscle compartment. The role of this layer is unclear.

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.

Morphological analysis of the hagfish heart. I. The ventricle, the arterial connection and the ventral aorta.

We have studied the heart in three species of hagfish: Myxine glutinosa, Eptatretus stoutii, and Eptatretus cirrhatus and report about the morphology of the ventricle, the arterial connection and the ventral aorta. On the whole, the hagfish heart lacks outflow tract components, the ventricle and atrium adopt a dorso-caudal rather than a ventro-dorsal relationship, and the sinus venosus opens into the left side of the atrium. This may indicate a "defective" cardiac looping during embryogenesis. The ventral aorta is elongated in M. glutinosa and E. stoutii but sac-like in E. cirrhatus. The ventricles are entirely trabeculated. The myocytes show a low myofibrillar content and junctional complexes formed by fascia adherens and desmosomes. Gap junctions could not be demonstrated. Myocardial cells in M. glutinosa contain numerous lipid droplets. These droplets are less numerous in E. stoutii and practically absent in E. cirrhatus, suggesting different metabolic requirements. Other cell types present in the ventricle are chromaffin cells and granular leukocytes that contain rod-shaped granules. The ventricle-aorta connection is guarded by a bicuspid valve with left and right, pocket-like leaflets. The leaflets extend from the cranial end of the ventricle into the aorta but the junction is asymmetrical. This junction contains a ganglion-like structure in E. cirrhatus. The ventral aorta shows endothelial, media, and adventitial layers. The media contains smooth muscle cells surrounded by dense bands formed by tightly-packed extracellular filaments. In addition, a short number of elastic fibers are observed in M. glutinosa and E. stoutii. Cellular and extracellular elements are more loosely organized in the aorta of E. cirrhatus. The collagenous adventitia contains ganglion-like cells in the three species. In the absence of nerves, chromaffin and ganglion-like cells may control the activity of the myocardium and that of the aortic smooth muscle cells, respectively.

The structure of the gas bladder of the spotted gar, Lepisosteus oculatus.

We report here on the macroscopic, light microscopic, and electron microscopic structure of the gas bladder (GB) of the spotted gar, Lepisosteus oculatus. The GB opens into the pharynx, dorsal to the opening of the oesophagus, through a longitudinal slit bordered by two glottal ridges. Caudal to the ridges, the GB is an elongated sac divided into a central duct and right and left lobes. The lobes are formed by a cranio-caudal sequence of large air spaces that open into the central duct. The structure of the GB is that of a membranous sac supported by a system of septa arising from the walls of a central duct. The septa contain variable amounts of striated and smooth muscle might function to maintain the bladder shape and in providing contractile capabilities. The presence of muscle cells, nerves, and neuroepithelial cells in the wall of the GB strongly suggests that GB function is tightly regulated. The central duct and the apical surface of the thickest septa are covered by mucociliated epithelium. Most of the rest of the inner bladder surface is covered by a respiratory epithelium which contains goblet cells and a single type of pneumocyte. These two cell types produce surfactant. The respiratory barrier contains thick areas with fibrillar material and cell prolongations, and thin areas that only contain basement membrane material between the capillary wall and the respiratory epithelium. Lungs and GBs share many anatomical and histological features. There appears to be no clear criterion for structural distinction between these two types of respiratory organs.

Lympho-granulocytic tissue associated with the wall of the spiral valve in the African lungfish Protopterus annectens.

We describe the structure of the lympho-granulocytic tissue associated with the wall of the spiral valve of the African lungfish Protopterus annectens. The study was performed under freshwater conditions and after 6 months of aestivation. The lympho-granulocytic tissue consists of nodes surrounded by reticular tissue. The nodes are formed by an outer and an inner component separated by a thin collagenous layer. The outer component is a reticular-like tissue that contains two types of granulocytes, developing and mature plasma cells and melanomacrophage centres (MMCs). The inner component, the parenchyma, contains a meshwork of trabeculae and vascular sinusoids and shows dark and pale areas. The dark areas contain diffuse lymphoid tissue, with a large number of mitoses and plasma cell clusters. The pale areas contain a small number of macrophages and lymphocytes. Macrophages and sinus endothelial cells are filled with haemosiderin granules and appear to form part of the reticuloendothelial system of the lungfish. The reticular tissue houses granulocytes, plasma cells and MMCs and might serve for the housing and maturation of cells of the white series. After aestivation, the nodes undergo lymphocyte depletion, the suppression of mitosis, granulocyte invasion and the occurrence of cell death. By contrast, few histological changes occur in the reticular tissue. Whereas the nodes appear to be involved in lymphocyte proliferation and plasma cell maturation, the function of the reticular tissue remains obscure.

Structural analysis of chordae tendineae in degenerative disease of the mitral valve.

BACKGROUND: Degenerative disease of the mitral valve (DDMV) is always accompanied by lengthening and/or rupture of chordae tendineae. However, the mechanisms and the mode of chordal rupture remain controversial, and the pathologic anatomy of the apparently healthy chordae has mostly been overlooked. We analyze the structural aspects of both ruptured and intact chordae tendineae in DDMV. METHODS AND RESULTS: Structural and ultrastructural microscopic analyses indicate that both the extracellular matrix and the interstitial cells are severely affected. Degenerative chordae show alterations in the synthesis and deposition of collagen and elastin, disorganization of collagen bundles and rupture of collagen fibres, accumulation of proteoglycans and of cellular and vesicular remnants, and cell transformation into a myofibroblast phenotype. Structural disruption makes the spongiosa and the dense collagenous core separate and break. Degeneration of the chordae is segmental, affecting both chordae that are clearly abnormal, and chordae that appear healthy on visual inspection. CONCLUSIONS: Changes in both matrix synthesis and degradation disturb the ordered collagen arrangement and modify the structural and physical properties of the chordae. Progressive structural disruption of the diseased chordae is the cause of chordal rupture. Mitral surgery corrects the damage, but the underlying causes of DDMV are not corrected. Thus, progression of the disease and affectation of additional chordae may be at the basis of the late complications and the recurrent mitral regurgitation which occurs several years after surgery. Our results indicate that a more aggressive approach to surgery may be needed.

The spleen of the African lungfish Protopterus annectens: freshwater and aestivation.

We describe the structure of the spleen of the African lungfish Protopterus annectens in freshwater conditions, and after 6 months of aestivation. The spleen is formed by cortical tissue that surrounds the splenic parenchyma. The cortex is a reticulum that contains two types of granulocytes, developing and mature plasma cells, and melanomacrophage centres (MMCs). The parenchyma is divided into lobules that show a subcapsular sinus and areas of red pulp and white pulp. Red pulp contains vascular sinuses and atypical cords formed by delicate trabeculae. White pulp also contains vascular sinuses and cords. Structural data indicate that red pulp is involved in erythropoiesis, destruction of effete erythrocytes, and plasma cell differentiation. White pulp appears to be involved in the production of immune responses. Macrophages and sinus endothelial cells constitute the reticulo-endothelial system of the spleen. After aestivation, the number of MMCs increases, and spleen tissue is infiltrated by lymphocytes, granulocytes, and monocytes. Also, white pulp is reduced, and sinus endothelial cells undergo vacuolar degeneration. Lungfish spleen shares structural characteristics with secondary lymphoid organs of both ectothermic and endothermic vertebrates, but appears to have evolved in unique ways.

The alimentary canal of the African lungfish Protopterus annectens during aestivation and after arousal.

We describe the structural modifications that occur in the alimentary canal of the African lungfish Protopterus annectens during aestivation and after arousal. With fasting, all gut segments undergo structural modifications. The epithelium covering the intestinal vestibule undergoes bursts of activation at 4 months of aestivation, adopting a more quiescent appearance at 6 months. The ridge area of the spiral intestine shows, at 4 months of aestivation, epithelial disintegration, cell desquamation, cell death, and loss of the freshwater phenotype. Surprisingly, the epithelium adopts a stratified appearance at 6 months of aestivation. Except for epithelial disintegration, the smooth portion of the spiral intestine follows a similar pattern of modifications than the ridge area. The entire epithelium of spiral intestine appears to be renewed during aestivation. The presence of intraepithelial mast cells suggests that inflammation is part of the cellular response to aestivation. After arousal, cell phenotypes are restored in about 6 days, but full structural recovery is not attained during the experimental period (15 days post-aestivation). Several aspects of the cellular response to fasting are shared by a wide range of animal groups. This commonality agrees with the presence of a character that allows to adjust the structural and functional properties of the gut to food availability and food quality, and to the characteristics of the fasting episodes.

The gut of the juvenile African lungfish Protopterus annectens: a light and scanning electron microscope study.

We describe the microstructure of the alimentary canal of the juvenile lungfish Protopterus annectens. Following the oesophagus, the gut is formed by a long segment that extends down to the pyloric valve. This segment, classically named stomach, is lined by a transitional epithelium but lacks all characteristics of the vertebrate stomach. It has been defined here as the intestinal vestibule. The spiral valve is divided into a first large chamber, which contains mucosal ridges, and a second smooth portion. The entire spiral valve is lined with a pseudostratified columnar epithelium that contains approximately six cell types: enterocytes, goblet cells, ciliated cells, leukocytes, dark pigment cells, and vascular cells. Enterocytes and goblet cells show a high number of cytoplasmic vacuoles. The number and size of the vacuoles, and the number of ciliated cells, decreases from the anterior toward the posterior end, suggesting that most of the digestive processes take place in the anterior part of the spiral valve. The epithelium overlies a lamina propria in the first large chamber and a vascular plexus in the smooth portion. The cloaca has a thick muscular wall covered by a transitional epithelium. An extensive lymphatic system formed by capillaries and lymphatic micropumps is present along the entire wall of the alimentary canal.

The atrioventricular region of the teleost heart. A distinct heart segment.

This article reports the morphological characteristics of the atrioventricular (AV) region of the teleost heart. A total of 29 teleost species belonging to 19 families and 10 orders were analyzed. The study includes hearts with entirely trabeculated ventricles and hearts possessing a ventricular compacta. In all cases, the AV region is formed by a ring of compact myocardium surrounded by a connective tissue ring. The myocardium contains vessels in most species having entirely trabeculated ventricles, and in all species possessing a compacta. The ring of connective tissue is rich in collagen and elastin, partially isolating the AV muscle from the surrounding musculature. Continuity between the AV muscle and the atrial and ventricular musculature is always observed. The AV muscle supports the AV valves, which are formed by two leaflets, lack papillary muscles, show a collagenous fibrosa on the atrial side, and present a variable number of cells and a variable amount of extracellular material. Ventricular trabeculae attach to the AV muscle. These trabeculae may help to bear the stress generated by ventricular contraction and may constitute a preferential pathway for the conduction of electrical impulses. In all teleosts studied, the AV region can be recognized as a distinct morphological segment interposed between the atrium and the ventricle. The morphological characteristics of the AV segment appear to be species specific.

The anatomy of the gastrointestinal tract of the African lungfish, Protopterus annectens.

The gastrointestinal tract of the African lungfish Protopterus annectens is a composite, which includes the gut, the spleen, and the pancreas. The gut is formed by a short oesophagus, a longitudinal stomach, a pyloric valve, a spiraling intestine, and a cloaca. Coiling of the intestine begins dorsally below the pylorus, winding down to form six complete turns before ending into the cloaca. A reticular tissue of undisclosed nature accompanies the winding of the intestinal mucosa. The spleen is located along the right side of the stomach, overlapping the cranial end of the pancreas. The pancreas occupies the shallow area, which indicates on the gut dorsal side the beginning of the intestine coiling. In addition, up to 25 lymphatic-like nodes accompany the inner border of the spiral valve. The mesenteric artery forms a long axis for the intestine. All the components of the gastrointestinal tract are attached to each other by connective sheaths, and are wrapped by connective tissue, and by the serosa externally. We believe that several previous observations have been misinterpreted and that the anatomy of the lungfish gut is more similar among all the three lungfish genera than previously thought. Curiously, the gross anatomical organization is not modified during aestivation. We hypothesize that the absence of function is accompanied by structural modifications of the epithelium, and are currently investigating this possibility.

The development of the epicardium in the sturgeon Acipenser naccarii.

This article reports on the development of the epicardium in alevins of the sturgeon Acipenser naccarii, aged 4-25 days post-hatching (dph). Epicardial development starts at 4 dph with formation of the proepicardium (PE) that arises as a bilateral structure at the boundary between the sinus venosus and the duct of Cuvier. The PE later becomes a midline organ arising from the wall of the sinus venosus and ending at the junction between the liver, the sinus venosus and the transverse septum. This relative displacement appears related to venous reorganization at the caudal pole of the heart. The mode and time of epicardium formation is different in the various heart chambers. The conus epicardium develops through migration of a cohesive epithelium from the PE villi, and is completed through bleb-like aggregates detached from the PE. The ventricular epicardium develops a little later, and mostly through bleb-like aggregates. The bulbus epicardium appears to derive from the mesothelium located at the junction between the outflow tract and the pericardial cavity. Strikingly, formation of the epicardium of the atrium and the sinus venosus is a very late event occurring after the third month of development. Associated to the PE, a sino-ventricular ligament develops as a permanent connection. This ligament contains venous vessels that communicate the subepicardial coronary plexus and the sinus venosus, and carries part of the heart innervation. The development of the sturgeon epicardium shares many features with that of other vertebrate groups. This speaks in favour of conservative mechanisms across the evolutionary scale.

The structural characteristics of the heart ventricle of the African lungfish Protopterus dolloi: freshwater and aestivation.

This paper reports on the structure and ultrastructure of the ventricular myocardium of the African lungfish Protopterus dolloi in freshwater (FW), in aestivation (AE), and after the AE period. The myocardium shows a conventional myofibrillar structure. All the myocytes contain large intracytoplasmic spaces occupied by a pale material that could contain glycosaminoglycans and/or glycogen, which may be used as food and water reservoirs. In FW, the myocytes in the trabeculae associated with the free ventricular wall show structural signs of low transcriptional and metabolic activity (heterochromatin, mitochondria of the dense type). These signs are partially reversed during the AE period (euchromatin, mitochondria with a light matrix), with a return to the FW appearance after arousal. The myocytes in the septum show, in FW conditions, nuclear polymorphism (heterochromatin, euchromatin), and two types (colliquative and coagulative) of necrosis. In AE, all the septal myocytes show euchromatin, and the number of necrotic cells increases greatly. Cell necrosis appears to be related to the septal architecture. After arousal, the septal myocytes exhibit a heterochromatin pattern, the number of necrotic cells decreases, cell debris accumulates under the endocardium, and phagocytosis takes place. Despite being a morphologic continuum, the trabeculae associated with the free ventricular wall appear to constitute a different compartment from that formed by the trabeculae in the ventricular septum. Paradoxically, AE appears to trigger an increase in transcriptional and synthetic myocardial activities, especially at the level of the ventricular septum. This activity may be involved in mechanisms of autocrine/paracrine regulation. Aestivation cannot be regarded as the result of a general depression of all cellular and organic activities. Rather, it is a much more complex state in which the interplay between upregulation and downregulation of diverse cell activities appears to play a fundamental role.

The heart of Sparus auratus: a reappraisal of cardiac functional morphology in teleosts.

This morphodynamic study provides an insight on how the architecture of the heart ventricle of the gilthead seabream (Sparus auratus) is designed to accomplish the functional performance typical of an active teleost species. Using an in vitro working heart preparation, mechanical performance was analyzed under loading (i.e., preload and afterload) challenges. The hearts were very sensitive to filling pressure increases. Maximum cardiac output (CO: 55.66+/-4.54 ml/min/kg body weight; mean+/-SEM) and maximum stroke volume (VS: 0.42+/-0.027 ml/kg body weight; mean+/-SEM) were obtained at an input pressure of 1 kPa. When exposed to output pressure (OP) changes, the hearts maintained constant CO and SV up to about 4 kPa; further increases of afterload significantly compromised mechanical performance. Surprisingly, this "athletic" pumping performance was achieved by an entirely trabeculated pyramidal ventricle. The ventricular architecture was characterized by a system of small luminae and trabecular sheets radiating outward from the central lumen. The most peripheral part of the ventricular chamber contained single trabeculae and the corresponding lacunary spaces. The ventricular cavity was bounded by an outer myocardial monolayer "shell" to which the peripheral trabeculae were attached. Myofibril organization differed in the trabeculae and in the outer monolayer. The structural features challenge common beliefs regarding the typical "athletic" teleost heart design.

Ventricle and outflow tract of the African lungfish Protopterus dolloi.

We report a morphologic study of the heart ventricle and outflow tract of the African lungfish Protopterus dolloi. The ventricle is saccular and appears attached to the anterior pericardial wall by a thick tendon. An incomplete septum divides the ventricle into two chambers. Both the free ventricular wall and the incomplete ventricular septum are entirely trabeculated. Only a thin rim of myocardium separates the trabecular system from the subepicardial space. The outflow tract consists of proximal, middle, and distal portions, separated by two flexures, proximal and distal. The proximal outflow tract portion is endowed with a layer of compact, well-vascularized myocardium. This portion is homologous to the conus arteriosus observed in the heart of most vertebrates. The middle and distal outflow tract portions are arterial-like, thus being homologous to the bulbus arteriosus. However, the separation between the muscular and arterial portions of the outflow tract is not complete in the lungfish. A thin layer of myocardium covers the arterial tissue, and a thin layer of elastic tissue underlies the conus myocardium. Two unequal ridges composed of loose connective tissue, the spiral and bulbar folds, run the length of the outflow tract. They form an incomplete division of the outflow tract, but fuse at the distal end. The two folds are covered by endocardium and contain collagen, elastin, and fibroblast-like cells. They appear to be homologous to the dextro-dorsal and sinistro-ventral ridges observed during the development of the avian and mammalian heart. Two to three rows of vestigial arterial-like valves appear in the dorsal and ventral aspects of the conus. These valves are unlikely to have a functional role. The possible functional significance of the "gubernaculum cordis," the thick tendon extending between the anterior ventricular surface and the pericardium, is discussed.

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.

The development of the sturgeon heart.

This paper presents a sequential analysis of the development of the sturgeon (Acipenser naccarii) heart from the end of gastrulation to the early juvenile stages. At late neurulation, the heart appears as a straight, short tube located over the endoderm that forms the wall of the yolk sac, in front of the developing head. The heart axis is aligned with the axis of the developing head. Subsequently, the heart elongates and adopts a C-shape, and its axis becomes perpendicular to that of the head. Around the time of hatching, the heart loses the loop and appears as a mostly straight tube with the chambers arranged in a craniocaudal sequence: outflow tract, ventricle, atrium, and a small sinus venosus. During the first 4 days post-hatching (dph), the heart starts looping again, adopts a C-shape, and undergoes a counterclockwise movement that brings the atrium to the left of the outflow tract and the ventricle to a caudal position. Thus, a primary and a secondary cardiac loop occur in the sturgeon. Later, the atria come to occupy a middle position behind the outflow tract, and the sinus venosus shifts from a caudal to a dorsal position. A morphological arrangement similar to that found in adult sturgeons is attained in all specimens at days 7-9 dph. The external changes are accompanied by a series of internal modifications that include trabeculation (3-4 dph), development of endocardial cushions in the atrioventricular canal (4 dph) and in the conus arteriosus (3-4 dph), conus (22-24 dph) and atrioventricular (18-20 dph) valve formation, and development of the epicardium (4 dph) and the coronary vessels (10 dph). The main developmental features of the heart have been registered, and a basic body of information, which should be very useful in future developmental studies, has been established. Similarities and dissimilarities between the development of the sturgeon heart and that of other vertebrates are underscored.

Differentiation of the cardiac outflow tract components in alevins of the sturgeon Acipenser naccarii (Osteichthyes, Acipenseriformes): implications for heart evolution.

Previous work showed that in the adult sturgeon an intrapericardial, nonmyocardial segment is interposed between the conus arteriosus of the heart and the ventral aorta. The present report illustrates the ontogeny of this intermediate segment in Acipenser naccarii. The sample studied consisted of 178 alevins between 1 and 24 days posthatching. They were examined using light and electron microscopy. Our observations indicate that the entire cardiac outflow tract displays a myocardial character during early development. Between the fourth and sixth days posthatching, the distal portion of the cardiac outflow tract undergoes a phenotypical transition, from a myocardial to a smooth muscle-like phenotype. The length of this region with regard to the whole outflow tract increases only moderately during subsequent developmental stages, becoming more and more cellularized. The cells soon organize into a pattern that resembles that of the arterial wall. Elastin appears at this site by the seventh day posthatching. Therefore, two distinct components, proximal and distal, can be recognized from the fourth day posthatching in the cardiac outflow tract of A. naccarii. The proximal component is the conus arteriosus, characterized by its myocardial nature and the presence of endocardial cushions. The distal component transforms into the intrapericardial, nonmyocardial segment mentioned above, which is unequivocally of cardiac origin. We propose to designate this segment the "bulbus arteriosus" because it is morphogenetically equivalent to the bulbus arteriosus of teleosts. The present findings, together with data from the literature, point to the possibility that cells from the cardiac neural crest are involved in the phenotypical transition that takes place at the distal portion of the cardiac outflow tract, resulting in the appearance of the bulbus arteriosus. Moreover, they suggest that the cardiac outflow tract came to be formed by a bulbus arteriosus and a conus arteriosus from an early period of the vertebrate evolutionary story. Finally, we hypothesize that the embryonic truncus of birds and mammals is homologous to the bulbus arteriosus of fish.

The structure of the conus arteriosus of the sturgeon (Acipenser naccarii) heart: II. The myocardium, the subepicardium, and the conus-aorta transition.

Sturgeons constitute a family of living "fossil" fish whose heart is related to that of other ancient fish and the elasmobranches. We have undertaken a systematic study of the structure of the sturgeon heart aimed at unraveling the relationship between the heart structure and the adaptive evolutionary changes. In a related paper, data were presented on the conus valves and the subendocardium. Here, the structure of the conus myocardium, the subepicardial tissue, and the conus-aorta transition were studied by conventional light, transmission, and scanning electron microscopy. In addition, actin localization by fluorescent phalloidin was used. The conus myocardium is organized into bundles whose spatial organization changes along the conus length. The variable orientation of the myocardial cell bundles may be effective in emptying the conus lumen during contraction and in preventing reflux of blood. Myocardial cell bundles are separated by loose connective tissue that contains collagen and elastin fibers, vessels, and extremely flat cells separating the cell bundles and enclosing blood vessels and collagen fibers. The ultrastructure of the myocardial cells was found to be very similar to that of other fish groups, suggesting that it is largely conservative. The subepicardium is characterized by the presence of nodular structures that contain lympho-hemopoietic (thymus-like) tissue in the young sturgeons and a large number of lymphocytes after the sturgeons reach sexual maturity. This tissue is likely implicated in the establishment and maintenance of the immune responses. The intrapericardial ventral aorta shows a middle layer of circumferentially oriented cells and internal and external layers with cells oriented longitudinally. Elastin fibers completely surround each smooth muscle cell, and the spaces between the different layers are occupied by randomly arranged collagen bundles. The intrapericardial segment of the ventral aorta is a true transitional segment whose structural characteristics are different from those of both the conus subendocardium and the rest of the ventral aorta.

Structure of the conus arteriosus of the sturgeon (Acipenser naccarii) heart. I: the conus valves and the subendocardium.

Sturgeons are bony fish that retain structural traits typical of the more primitive Chondrostei. From an evolutionary viewpoint, sturgeons are considered relic fish. However, they show remarkable ecological plasticity and are well adapted to contemporary environmental conditions. Although development of the cardiovascular system is critical for all organs and systems, and is affected by evolutionary changes, the structure of the sturgeon heart has been mostly overlooked. This is also true for the conus arteriosus, which, as in Chondrostei, is endowed with several rows of valves and a layer of contractile myocardium. This work reports on the structure of the valves, the endocardium, and the subendocardium of the conus arteriosus of the sturgeon (Acipenser naccarii) heart. It is part of a broader study that aims to cover the entire structure of the sturgeon heart. The conus arteriosus of 15 A. naccarii hearts, ranging in age from juveniles to sexually-differentiated adults, has been studied by conventional light, transmission (TEM), and scanning electron microscopy (SEM). In addition, maceration of the soft tissues with NaOH, and actin localization by fluorescent phalloidin has been used. The conus is a tubular chamber that arises from the right ventricular side and presents two constrictions at the conus-ventricle and conus-aorta junctions. The conus is endowed with three rows of valves: one distal and two proximal. The segment of the conus located between the distal and the two proximal rows is devoid of valvular structures. The distal row has four leaflets, while the two proximal rows show the greatest variation in leaflet number, size, and shape. All leaflets have collagenous chordae tendineae arising from the free border and from the parietal side of the leaflets. The endocardium is a flat endothelium which shows a thick, irregular basement membrane. The leaflet body is formed by a loose connective tissue which blends with the subendocardium. The subendocardium is a connective tissue consisting of myofibroblasts, collagen, and elastin. It is divided into two distinct areas: one proximal, which shows little elastin and poorly organized collagen; and one distal, which is rich in elastin, with cells and extracellular fibers organized into layers that are oriented in alternative circumferential and longitudinal directions. The present report is the first systematic analysis of the structure of the sturgeon conus. Descriptions of the conus valves should recognize the existence of three valve rows only. The variability in valve morphology, and the loose structure of the leaflet tissue make it unlikely that the valves play an effective role in preventing blood backflow. In this regard, the ventricle-conus constriction may act as a sphincter. The subendocardium is an elastic coat capable of actively sustaining the tissue deformation that accompanies the heart contractile cycle. Further comparative studies are needed to provide deeper insight into the structural changes that accompany phyletic diversification.