Cardiac morphodynamic remodelling in the growing eel (Anguilla anguilla L.).

The morphodynamic changes occurring during growth were evaluated in the eel (Anguilla anguilla L.) heart. Using an in vitro working heart preparation, cardiac performance of small (body mass 96.76 +/- 27.49 g; mean +/- s.d.) and large (body mass 656 +/- 12 g; mean +/- s.d.) eels was compared under basal conditions and under loading (i.e. preload and afterload) challenges. A parallel morphometric evaluation of the ventricle was made using light and transmission electron microscope images. The small eel hearts show a basal cardiac output lower than their large counterparts (heart rate fh, 38.93 +/- 2.82 and 52.7 +/- 1.8 beats min(-1), respectively; stroke volume Vs, 0.27 +/- 0.017 and 0.37 +/- 0.016 ml kg(-1), respectively; means +/- s.e.m.). The two groups show similar responses at increasing preload, but differ remarkably at increasing afterload. Small eel hearts decreased Vs at afterload greater than 3 kPa, in contrast to larger hearts, which maintained constant Vs up to 6 kPa. These changes in mechanical performance are related to structural differences. Compared with the small eels, the large eels show an increase in the compacta thickness and in the diameter of the trabeculae in the spongiosa, together with reduction of the lacunary spaces. The increased compacta thickness is attained by enlargements of both the muscular and vascular compartments and reduction of the interstitium; consequently, this layer appears more compacted. Both compacta and spongiosa show higher number of myocytes together with reduced cross-sectional area and myofibrillar compartment. The compacta also shows an increased mitochondrial compartment. Our results document a cardiac morphodynamic remodelling in the growing eel.

The conus valves of the adult gilthead seabream (Sparus auratus).

The conus (bulbo-ventricular) valves of teleosts perform a key function in the control of blood backflow during ventricular diastole. However, the structural characteristics of these valves are almost unknown. This paper presents a systematic anatomical, histological and structural study of the conus valves of the adult gilthead seabream (Sparus auratus). S. auratus shows two major left and right valves consisting of the leaflet and the supporting sinus. Each valvar leaflet can be divided into a stout proximal body and a flap-like distal region. The proximal body is structured into three layers: a luminal fibrosa, a dense cellular core and a parietal fibrosa. The luminal fibrosa is a collagenous structure extending the entire length of the leaflet, while the parietal fibrosa is restricted to the most proximal area. The dense cellular core consists of fibroblastic cells and a matrix rich in glycoconjugates, collagen and elastin. The histochemical and structural data suggest that the luminal fibrosa bears most of the force associated with valvar closure, while the cellular core acts as a cushion dampening vibrations and absorbing the elastic recoil. The sinus wall is a fibrous layer which shows proximal-distal differences in thickness. It also shows compositional differences that can be related to mechanical function. We describe the presence of a fibrous cylinder formed by the sinus wall, the fibrous interleaflet triangles and the fibrous layer that covers the inner surface of the conus myocardium. This fibrous cylinder constitutes the structural nexus between the ventricle, the conus and the bulbus arteriosus, provides support for the conus valves and separates the valvar complex from the surrounding tissues. The structure of the conus valves in S. auratus is different from that found in other vertebrates. Anatomical similarities between the conus valves and the mammalian arterial valves are emphasized. Each phyletic group appears to have developed specific structures in order to perform similar functions.

The conus arteriosus of the adult gilthead seabream (Sparus auratus).

This paper reports on the presence of the conus arteriosus in the heart of the adult gilthead seabream, Sparus auratus (Perciformes, Teleostei). The junctional region between the single ventricle and the bulbus arteriosus has been studied by conventional light microscopy, and by scanning and transmission electron microscopy. In addition, fluorescent phalloidin and antibodies against the muscle myosin heavy chains, laminin and collagen type IV have been used. The conus arteriosus is a distinct muscular segment interposed between the ventricle and the bulbus arteriosus. It is clearly different from the bulbus arteriosus due to its myocardial nature. It can also be distinguished from the ventricular myocardium because: (1) it has a conus shape; (2) it is formed by compact, well-vascularized myocardium; (3) it is surrounded on its inner and outer faces by fibrous layers rich in collagen and elastin; (4) it constitutes the anatomical support of the so-termed conus valves; (5) it shows intense staining for laminin and type-IV collagen; and (6) the myocardial cells located close to the inner fibrous layer are helicoidally arranged. By contrast, the ventricular myocardium is highly trabecular, lacks a compacta, shows no vessels, and presents barely detectable amounts of laminin and collagen type IV. The presence of a distinct conus arteriosus in the heart of an evolutionary advanced teleost species indicates that the conus is not a vestigial segment from the evolutionary or embryological points of view. The characteristic spatial arrangement of the conus myocytes strongly suggests that the conus is implicated in the mechanical performance of the conus valves.

Origin and course of the coronary arteries in normal mice and in iv/iv mice.

This paper reports on the origin and distribution of the coronary arteries in normal mice and in mice of the iv/iv strain, which show situs inversus and heterotaxia. The coronary arteries were studied by direct observation of the aortic sinuses with the scanning electron microscope, and by examination of vascular corrosion casts. In the normal mouse, the left and right coronaries (LC, RC) arise from the respective Valsalva sinus and course along the ventricular borders to reach the heart apex. Along this course the coronary arteries give off small branches at perpendicular or acute angles to supply the ventricles. The ventricular septum is supplied by the septal artery, which arises as a main branch from the right coronary. Conus arteries arise from the main coronary trunks, from the septal artery and/or directly from the Valsalva sinus. The vascular casts demonstrate the presence of intercoronary anastomoses. The origin of the coronary arteries was found to be abnormal in 84% of the iv/iv mice. These anomalies included double origin, high take-off, slit-like openings and the presence of a single coronary orifice. These anomalies occurred singly or in any combination, and were independent of heart situs. The septal artery originated from RC in most cases of situs solitus but originated predominantly from LC in situs inversus hearts. Except for this anomalous origin no statistical correlation was found between the coronary anomalies and heart situs or a particular mode of heterotaxia. The coronary anomalies observed in the iv/iv mice are similar to those found in human hearts. Most coronary anomalies appear to be due to defective connections between the aortic root and the developing coronaries. iv/iv mice may therefore constitute a good model to study the development of similar anomalies in the human heart.

Light and electron microscopy of the bulbus arteriosus of the European eel (Anguilla anguilla).

The bulbus arteriosus of teleost fish acts as an elastic reservoir that dilates during ventricular systole to store a large part of the cardiac stroke volume. Despite its functional importance, the knowledge of the structure of the bulbus wall is still fragmentary. We have undertaken a series of studies in order to establish a general morphological plan of the teleost bulbus. The bulbus arteriosus of the European eel is studied here by means of conventional light, and transmission and scanning electron microscopy. The inner surface of the bulbus wall is irregular due to the presence of branching ridges that flatten and disappear toward the ventral aorta. The ridge surface is covered by flattened endocardial cells that show moderately dense bodies. In the ridge tissue, cells near the endocardium are mostly undifferentiated and appear isolated in a loose filamentous matrix. Ridge cells progressively cluster toward the middle layer, become surrounded by a dense matrix, and adopt characteristics typical of smooth muscle cells. This suggests the existence of a differentiation gradient. The middle layer is formed by typical smooth muscle cells embedded in a meshwork matrix that contains thin and thick filaments. Stretching of this meshwork suggests an active role of smooth muscle cells in bulbus wall dynamics. Furthermore, large areas of the extracellular space are occupied by elastin-like material. The amount of this material decreases toward the external layer. Collagen is demonstrated across the entire thickness of the bulbus wall, its amount and organization increasing from the inner toward the outer bulbus surface. The existence of matrix gradients should progressively increase wall strength, maintaining bulbus dilation within safe physiological parameters. The epicardium is formed by flattened cells that contain numerous pinocytotic vesicles, suggesting an active interchange of solutes with the pericardial cavity.

Bulbus arteriosus of the Antarctic teleosts. II. The red-blooded Trematomus bernacchii.

The structure of the bulbus arteriosus of the Antarctic teleost, Trematomus bernacchii, has been studied by light, scanning, and transmission electron microscopy. The wall of the bulbus arteriosus is divided into endocardial, subendocardial, middle and external layers. The endocardial endothelium covers the inner surface of the bulbus wall and invaginates into the subendocardium to form solid epithelial cords that show secretory activity. The subendocardial tissue is divided into finger-like ridges. Ridge cells located under the endocardium appear in niches limited by collagen fibers and thin cell extensions. Away from the endocardium ridge cells cluster into small groups, show some of the characteristics of smooth muscle cells, and appear enmeshed in a filamentous meshwork that lacks collagen and elastin fibers. The middle bulbus layer is formed by typical smooth muscle cells that are enmeshed in a filamentous meshwork similar to that observed in the ridges. The ridges and the middle layer appear to be formed by the same cell type, smooth muscle, with a gradient of differentiation from the endocardium toward the middle layer. In the absence of elastin fibers the filamentous meshwork should confer elastic properties to the bulbus wall. The stretching of the meshwork along the main axis of the middle layer cells, and between different cellular layers, suggests the existence of tensile stress and, hence, the involvement of smooth muscle cells in bulbus wall dynamics. The external layer is formed by numerous cellular types embedded in a collagenous matrix. Among these cellular types, myofibroblasts, macrophages, granulocytes, lymphocytes, dendrite-like cells, degenerating cells, and plasma cells can be recognized. The subepicardial tissue appears to be a specialized site involved in the production of the humoral immune response and displays many of the morphological characteristics of a germinal center. The outer limiting layer of the bulbus, the visceral pericardium, is formed by epithelial cells that show desmosomes and tight junctions. This suggests a close control of permeability with respect to the pericardial fluid.

Bulbus arteriosus of the antarctic teleosts. I. The white-blooded Chionodraco hamatus.

The bulbus arteriosus of teleost fish is a thick-walled chamber that extends between the single ventricle and the ventral aorta. The functional importance of the bulbus resides in the fact that it maintains a steady blood flow into the gill system through heart contraction. Despite of this, a thorough study of the structure of the bulbus in teleost fish is still lacking. We have undertaken a morphologic study of the bulbus arteriosus in the stenothermal teleosts of the Antarctic sea. The structural organization of the bulbus arteriosus of the icefish Chionodraco hamatus has been studied here by conventional light, scanning, and transmission electron microscopy. The inner surface of the bulbus shows a festooned appearance due to the presence of longitudinal, unbranched ridges that extend between the ventricle and the arterial trunk. The wall of the bulbus is divided into endocardial, subendocardial, middle, and external layers. Endocardial cells show a large number of moderately-dense bodies. The endocardium invaginates into the subendocardium forming solid epithelial cords that contain numerous secretory vacuoles. Cells in the subendocardium group into small domains, have some of the morphological characteristics of smooth muscle cells, and appear enmeshed in a three-dimensional network of matrix filaments. Cells in the middle layer are typical smooth muscle cells. They appear arranged into layers and are surrounded by a filamentous meshwork that excludes collagen fibers. Orientation of this meshwork occurs in the vicinity of the smooth muscle cells. Elastin fibers are never observed. The external layer is formed by wavy collagen bundles and fibroblast-like cells. This layer lacks blood vessels and nerve fibers. The endocardium and the endocardium-derived cords are secretory epithelia that may be involved in the formation ofmucins or glycosaminoglycans. These mucins may have a protecting effect on the endocardium. The subendocardium and the middle layer appear to be formed by the same cell type, smooth muscle, with a gradient of differentiation from the secretory (subendocardium) to the contractile (middle layer) phenotype. Despite the absence of elastin fibers, the filamentous matrix could maintain the elastic properties of the bulbus wall. Smooth muscle cells appear to be actively involved in bulbus wall dynamics. The restriction of collagen to the external layer suggests that it may control wall dilatation and bulbus compliance. When comparison was possible, structural differences between C. hamatus and temperate teleosts seemed to be not species-related, but of phenotypic adaptative significance. This is remarkable since Antarctic fishes have lived isolated in freezing waters for the last two million years.

Collagenous skeleton of the human mitral papillary muscle.

The papillary muscles (PM) of the heart have been the subject of numerous structural and functional studies. However, despite the importance of the collagenous compartment of the heart in the mechanical and electrical properties of the myocardium, little information is available on the structural organization of collagen within the PM. We study here the structural organization of collagen within the mitral papillary muscles (PM) of the human heart. Fragments of human mitral PM from normal and hypertensive subjects were macerated in NaOH to eliminate the cellular components. Macerated and nonmacerated samples were then studied with the scanning electron microscope (SEM). SEM shows that cardiac myocytes and endomysial capillaries are ensheathed in a layer of collagenous tissue. The myocyte sheath wall is formed by thin collagen fibers oriented at right angles to the main cell axis. These sheaths are open structures, collagen fibers continuing into adjacent sheaths at the points of lateral communications. Thick perimysial septa do not divide the PM tissue into separate compartments. Hypertensive hearts show perivascular and interstitial fibrosis. In addition, the lumen of the coronary vessels is reduced or obliterated, and large areas of the myocardium are substituted by densely packed collagen. Endomysial sheaths constitute a continuous collagenous layer that replicates the myocyte network. The endomysium should play a complex role in myocardial mechanics, assuring the equal distribution of force during the cardiac cycle. The absence of insulating boundaries should facilitate lateral propagation of excitation. Fibrosis in hypertensive hearts appears to be both reactive and reparative. The increase in the amount of collagen should greatly impair contractile capabilities and electrical conductance, severely compromise heart function, and contribute to development of heart failure.

Atrioventricular valves of the mouse: III. Collagenous skeleton and myotendinous junction.

BACKGROUND: The leaflet tissue of the mouse atrioventricular (AV) valves contains a system of wavy collagen bundles that organize like tendons, orientate along lines of tension, and constitute an essential component of the valve tissue. The organization of these bundles is different in the two AV valves, reflecting differences in the anatomy of the entire valvular complex. Further insights into this kind of organization are needed to gain a complete understanding of the functional anatomy of the mouse AV valves. METHODS: The endocardial covering of the mouse AV valves (from 21 days to 1 year of age) was eliminated by the sonication or the maceration method. This allowed us to study in situ the organization of the collagenous valve skeleton, as well as the structure of the myotendinous junction. RESULTS: The leaflets of the two AV valves are formed by a fibrous layer (on the ventricular side) and a spongy layer (on the atrial side). The fibrosa is formed by undulating collagen bundles that organize and orientate differently on the right and left sides. The spongiosa is formed, on both sides, by a loose network of thin collagen fibers with no apparent orientation. Myocardial cells in the papillary muscles of the tricuspid valve are elongated and show cone-shaped tips. Collagen fibers attach to the myocyte surface. Collagen struts and thin septa can also be recognized. On the other hand, the collagenous components of the mitral leaflets attach tangentially to the mitral papillary muscles. On the two sides, the myocytes appear to be ensheathed in a layer of collagenous tissue. The sheaths are formed by circularly arranged fibers and appear to be tightly interconnected. CONCLUSIONS: The differences in the collagenous organization between the two AV valves reflect differences in the gross anatomy of the valves. The attachment of collagen to the papillary myocytes in the tricuspid valve resembles that of a typical myotendinous junction. However, the collagen-muscle junction in the mitral valve is more similar to the structure of a pennate muscle. The collagen matrix of the heart has been divided into endomysial, perimysial, and epimysial components. The presence of sheaths housing individual myocytes and capillaries, struts, and thin septa, corresponds to the endomysium. The absence of perimysial septa, which aggregate myocytes into groups, is striking, but this may just be a species difference. The appropriateness of the term epimysium, as applied to the heart, is discussed.

Atrioventricular valves of the mouse: II. Light and transmission electron microscopy.

BACKGROUND: Mouse atrioventricular (AV) valves present a number of conspicuous morphologic differences with human AV valves. Given the existence of these differences, it is important to know the structural organization of mouse AV valves. Since the mouse is often considered to be a good animal model for developmental and anatomical studies, the presence of significant differences in structure may render comparative studies difficult. In addition, we wished to learn about the existence of structural changes in the mouse AV valves with age. METHODS: The structural organization of mouse AV valves from 21 days to 1 year of age was studied by polarizing microscopy and by conventional light and transmission electron microscopy. RESULTS: Polarizing microscopy reveals the presence of a system of birefringent fibers that consist of collagen bundles that organize like tendons. The spatial organization of these fibers is different in the two AV valves, reflecting differences in the anatomy of the entire valvular complex. Interstitial cells (IC) are of two different phenotypes: some are typical fibroblasts, while some others share smooth muscle cell characteristics. In addition, small areas of fibrocartilage are also observed. The compactness and thickness of the collagen bundles increase with age. Also with age, the basement membranes become thickened or multilayered, and matrix vesicles and deposits of amyloid P can be observed. CONCLUSIONS: The collagenous birefringent fibers form an internal skeleton that should transmit the cycling stress evenly over the entire leaflets. IC should help to maintain the structure and deformability of the valve tissue and appear actively involved in the synthesis and renewal of extracellular material. The cartilaginous foci appear to be a normal component of the valve tissue. The structural changes observed in old animals appear to be related to the degenerative processes which take place in normal valvular tissues with age. Despite the structural differences, age changes appear to be similar in the AV valves of mouse and man.

The atrioventricular valves of the mouse. I. A scanning electron microscope study.

This paper reports a scanning electron microscope study of the morphology of the atrioventricular (AV) valves in the mouse. The leaflet tissue of the 2 AV valves consists of a continuous veil that shows no commissures or clefts. In all instances, the chordae that arise from the papillary system merge with the free border of the leaflet tissue. No distinct terminations of chordae were observed on the ventricular face of the valves. The leaflet tissue of the right AV valve can be divided into parietal and septal components on the basis of the insertion into the ventricular wall and of the papillary system. While the septal component is similar in shape, location and tension apparatus to the septal tricuspid leaflet in man, the parietal component appears to correspond to the anterior and posterior human leaflets. This segment of the valve is served by 3 papillary muscles that arise from the septal wall. The right AV valve is not a tricuspid structure from the morphological standpoint, but appears to function as such because of the particular attachment of the papillary muscles. The leaflet tissue of the mitral valve is served by 2 papillary muscles, anterior and posterior, which consist of muscular trabeculae extending from the heart apex to the base of the valve. These muscles remain associated with the ventricular wall. The leaflet tissue attaches directly to these papillary muscles, which give rise to a very small number of slender chordae. There are thus several important differences between the AV valves of the mouse and man.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of colchicine administration on the endothelial cells of the developing semilunar heart valves of the chick embryo.

Recent ultrastructural studies have revealed that differences exist in endothelial cell shape and cytoskeletal architecture between the arterial and ventricular faces of developing semilunar valves. In the present work we analyzed the morphologic response of the valvular endothelial cells of chick embryos to colchicine by light microscopy, scanning electron microscopy and transmission electron microscopy. The results show that colchicine administration during the stages of valve morphogenesis causes a very conspicuous disruption of the endothelial layer of the arterial face of the valves. The cells appear rounded and show massive surface blebbing. These alterations were not present in the endothelial cells on the ventricular face of the valves at the same stages. On the basis of these results we suggest that a difference in the degree of cell differentiation exists between the endothelial cells of the arterial and ventricular faces of the cusps and that this difference may have morphogenetic significance.

Retinal cell death occurs in the absence of retinal disc invagination: experimental evidence in papaverine-treated chicken embryos.

In an attempt to clarify the relationship between the presence of retinal cell death and the invagination of the optic vesicle, we have tested the occurrence and cytological characteristics of the retinal necrotic areas in the embryonic chicken after the administration in ovo of papaverine. Papaverine, a Ca2+ antagonist, was found to prevent the invagination of the optic vesicle. All embryonic retinae presented two distinct necrotic areas. However, these areas of cell death appeared abnormally located in the experimental, uninvaginated retina. One area was located at the transition between the retinal disc and the ventral wall of the optic vesicle; a second area was located in the dorsal wall of the optic vesicle, close to the optic stalk. We suggest that these necrotic areas represent the normal necrotic areas, should the invagination of the retinal disc have taken place. Retinal cell death appears to be programmed; it occurs whether the retinal disc invaginates or not. Cell death appears, in this experimental model, as a natural marker giving evidence that the embryonic retinal cells move from the optic stalk into the invagination retinal disc during normal eye cup formation. In addition to the uninvaginated optic vesicle the lens placode failed to invaginate in 45% of the cases, forming a lens vesicle in 55% of the remaining cases. This suggests that the two processes of invagination are governed by a different set of factors.

Vascular regression during the formation of the free digits in the avian limb bud: a comparative study in chick and duck embryos.

The pattern and structure of the blood vessels of the interdigital spaces of the leg bud have been studied by means of Indian ink injections and transmission electron microscopy in the chick and duck embryos. The results show that in the chick the interdigital necrotic process responsible for the freeing of the digits is followed by regression of the blood vessels. In the webbed foot of the duck, the interdigital necrotic processes are not followed by vascular regression. Transmission electron microscopic studies show that both in the chick and in the duck, interdigital blood vessels are immature structures lacking basal lamina. Dead cells of presumably endothelial origin were detected in the lumen of the regressing blood vessels of the chick but not in the duck. However, the intensity of this cell death process does not appear to be high enough to account by itself for the disappearance of the interdigital blood vessels. The possible relationships between interdigital mesenchymal cell death and vascular regression are discussed.

The surface anatomy of the human aortic valve as revealed by scanning electron microscopy.

The anatomy of the human aortic valve was studied by SEM in 36 subjects without cardiac pathology who had died of various accidental causes. Villous and lamellar tissue excrescences were observed at the node of Arantius and at the limit between the lunules and the load-bearing portion of the leaflets. The morphology of these structures suggests that they represent areas in which valve tissue becomes detached into the bloodstream. Fenestrations were present in the lunules of 14 specimens, with a higher incidence in specimens from subjects who were middle-aged or older. Our observations suggest that fenestrations appear initially as small perforations which then coalesce to form larger apertures. Two main types of endothelial cells, elongated and polygonal were detected on the endothelial surface of the leaflets. Both types of cells display a constant mode of arrangement on the different segments of the leaflets (lunules, node of Arantius and load-bearing portion of the leaflet). The possible relationships between endothelial cell morphology and the pattern of mechanical stress to which the leaflets are subjected is discussed.

Cell death in the dorsal part of the chick optic cup. Evidence for a new necrotic area.

The spatiotemporal pattern of morphogenetic cell death during the early development of the chick retina was studied by means of the neutral red vital staining and light microscopy. A modification of the conventional procedure of vital staining, which consisted of the injection of the dye into the neural tube lumen, was used for this purpose. In addition to the two areas of cell death known from previous literature, the first located in the ventral part of the optic cup and the second located in the insertion of the optic stalk with the diencephalon, a new area of cell death was described. This third necrotic area was located in the protruding dorsal part of the optic cup rim and was present throughout the stages 15 to 18. The area consisted of dying cells, fragments and phagocytosed cells. We suggest that this dorsal area of cell death could stop the intense dorsal growth of the optic cup and/or reshape the optic cup rim. Moreover, this area may influence the production of cell degeneration in the dorsal part of the invaginating lens placode.

The mechanisms of cell death and phagocytosis in the early chick lens morphogenesis: a scanning electron microscopy and cytochemical approach.

The cell surface characteristics of degenerating cells and phagocytes, as well as the participation of lysosomes in the cell death process associated with the early embryogenesis of chick lens rudiment, were studied by means of scanning electron microscopy and cytochemically using the Gomori-beta-glycerophosphate method for acid phosphatase. The prospective dying columnar epithelial cells lose their apical and basal processes and become rounded. The rounded, isolated, dying cells initially show a rough surface with some cytoplasmic constrictions followed by progressive break-up into several pitted fragments. Coincident with the loss of the columnar cell shape, acid phosphatase is localized within the Golgi apparatus and autophagic vacuoles which progressively increase in size. In contrast, the isolated dying cells and fragments do not show significant acid phosphatase activity. The role of lysosomes in this degenerative process is discussed. Neighboring epithelial cells phagocytose the dead cell fragments, becoming nonspecialized phagocytes. These consist of columnar epithelial cells and free cells which have migrated from the lens epithelium. Two mechanisms of internalization are observed. The most frequent mechanism takes place in both the columnar epithelial cells and the free cells, and consists of the progressive engulfment of the fragments into craters of the cell surface. The other mechanism is only detected in the free cells and takes place by pseudopod engulfment. We suggest that both phagocytic procedures could be related to the degree of intercellular connection. The presence of phagocytic internalization by crater formation in the epithelial cells could be a mechanism preserving the epithelial stability, which is necessary for a normal morphogenesis. Small microprocesses binding the surface of the phagocyte and the fragment are present prior to the internalization process. In the lens stalk and in the space located between the ectoderm and the lens vesicle, there are some cells displaying migratory characteristics. This fact suggests that an active migration of epithelial cells from the lens stalk could account for the process of detachment of lens vesicle from the ectoderm. The free cells appear to undergo an in situ progressive degeneration.

Changes in the endothelial morphology of the developing semilunar heart valves. A TEM and SEM study in the chick.

In view of recent evidence showing that shape and orientation of endothelial cells is determined by blood flow, the endothelium of the semilunar valves was studied in the developing chick heart using transmission and scanning electron microscopy. The results reveal significant developmental modifications of endothelial morphology and structure. These modifications can be linked to modifications of local blood flow and can also explain several aspects of valvular morphogenesis. The results substantially support the hypothesis of an involvement of hemodynamics in the development of the semilunar valves.

Malformations of the semilunar valves produced in chick embryos by mechanical interference with cardiogenesis. An experimental approach to the role of hemodynamics in valvular development.

The purpose of the present work was to analyze the role of hemodynamics in the morphogenesis and histogenesis of the semilunar valves. To achieve this goal we have studied the development of the chick semilunar valves in conditions of abnormal local flow. To obtain an abnormal pattern of local flow we have induced alterations of the cardiac septation process by mechanical interference of the development of the conus cordis. The malformations obtained by this procedure consisted of a spectrum of alterations in the process of incorporation of the aortic conus into the left ventricle. These malformations ranged from a simple widening of the outflow tract of the left ventricle to severe forms of double-outlet right ventricle and ventricular septal defects. Malformations of the semilunar valves consisting of extensive thickening of the leaflets and lack of maturation of the valve tissues were very often present in the malformed hearts. The malformation of the valve leaflets was more frequent and severe in the aortic valve at more advanced stages of development and in the hearts showing more severe alteration of the septation process. The absence of alterations in the semilunar valves of the control embryos and in the experimental embryos without alteration of the cardiac septation suggest a close relationship between the semilunar valves anomalies and the hemodynamic alterations present in the malformed hearts.