Nkx2.5‐negative myocardium of the posterior heart field and its correlation with podoplanin expression in cells from the developing cardiac pacemaking and conduction system

Recent advances in the study of cardiac development have shown the relevance of addition of myocardium to the primary myocardial heart tube. In wild‐type mouse embryos (E9.5–15.5), we have studied the myocardium at the venous pole of the heart using immunohistochemistry and 3D reconstructions of expression patterns of MLC‐2a, Nkx2.5, and podoplanin, a novel coelomic and myocardial marker. Podoplanin‐positive coelomic epithelium was continuous with adjacent podoplanin‐ and MLC‐2a‐positive myocardium that formed a conspicuous band along the left cardinal vein extending through the base of the atrial septum to the posterior myocardium of the atrioventricular canal, the atrioventricular nodal region, and the His‐Purkinje system. Later on, podoplanin expression was also found in the myocardium surrounding the pulmonary vein. On the right side, podoplanin‐positive cells were seen along the right cardinal vein, which during development persisted in the sinoatrial node and part of the venous valves. In the MLC‐2a‐ and podoplanin‐positive myocardium, Nkx2.5 expression was absent in the sinoatrial node and the wall of the cardinal veins. There was a mosaic positivity in the wall of the common pulmonary vein and the atrioventricular conduction system as opposed to the overall Nkx2.5 expression seen in the chamber myocardium. We conclude that we have found podoplanin as a marker that links a novel Nkx2.5‐negative sinus venosus myocardial area, which we refer to as the posterior heart field, with the cardiac conduction system. Anat Rec, 290:115–122, 2007. © 2006 Wiley‐Liss, Inc.     

Cardiac malformations in Pdgfrα mutant embryos are associated with increased expression of WT1 and Nkx2.5 in the second heart field

Platelet‐derived growth factor receptor alpha (Pdgfrα) identifies cardiac progenitor cells in the posterior part of the second heart field. We aim to elucidate the role of Pdgfrα in this region. Hearts of Pdgfrα‐deficient mouse embryos (E9.5–E14.5) showed cardiac malformations consisting of atrial and sinus venosus myocardium hypoplasia, including venous valves and sinoatrial node. In vivo staining for Nkx2.5 showed increased myocardial expression in Pdgfrα mutants, confirmed by Western blot analysis. Due to hypoplasia of the primary atrial septum, mesenchymal cap, and dorsal mesenchymal protrusion, the atrioventricular septal complex failed to fuse. Impaired epicardial development and severe blebbing coincided with diminished migration of epicardium‐derived cells and myocardial thinning, which could be linked to increased WT1 and altered α4‐integrin expression. Our data provide novel insight for a possible role for Pdgfrα in transduction pathways that lead to repression of Nkx2.5 and WT1 during development of posterior heart field–derived cardiac structures. Developmental Dynamics 239:2307–2317, 2010. © 2010 Wiley‐Liss, Inc.     

In normal development pulmonary veins are connected to the sinus venosus segment in the left atrium

Background: Classic theories descibe that the common pulmonary vein develops as an outgrowth from either the sinus venosus or atrial segment. Recent studies show that the pulmonary veins are connected to the sinu-atrial region before its differentiation into a sinus venosus and atrial segment. Methods: The development of the sinu-atrial region with regard to the developing common pulmonary vein and the growth of the atrial septum was investigated in avian embryos, using both scanning electron microscopy and immunohistochemistry. Embryos ranging between stage HH12 and HH28 were incubated with QH-1 that recognizes quail endothelial cells and precursors, HNK-1, that appears in this study to detect the myocardium of the sinus venosus, or with HHF-35, being specific for muscle actins. Also vascular casts of the heart were produced by injecting prepolymerized Mercox into the vascular system. Results: In preseptation stages the common pulmonary vein drains into the left part of the sinus venosus, that is clearly demarcated by the sinuatrial fold and HNK-1 expression. During atrial septation the left part of the sinus venosus, in contrast to the right part, loses its HNK-1 antigen from stage HH23 onwards, while at the same time the sinu-atrial fold in the left atrial dorsal wall flattens and disappears. From stage HH25 onwards HNK-1 expression is restricted to the right part of the sinus venosus, which contributes to the right atrium. The myocardial atrial septum never expresses the HNK-1 antigen, suggesting that the septum is of atrial origin. Discussion: It appeared that the sinus venosus does not only contribute to the sinus venarum of the right atrium, but also to the left atrium. © 1995 Wiley-Liss, Inc.     

人胚心静脉窦和传导系统的早期发育

目的 探讨早期人胚心静脉窦及传导系的发生发育机制. 方法 用抗α-平滑肌肌动蛋白(α-SMA)、抗α-横纹肌肌动蛋白(α-SCA)和抗结蛋白(DES)抗体对29例C10～C16期人胚心连续切片行免疫组织化学染色. 结果 人胚发育C12～C13期,系统静脉汇集形成的静脉窦出现于心包腔尾端原始横膈间充质中,静脉窦壁间充质细胞逐渐分化为α-SCA阳性的静脉窦心肌细胞.C14期,心包腔的扩张使静脉窦进入心包腔内,参与了右心房的形成.DES阳性传导系心肌的分化始于C10期心房室管右侧壁,随发育逐渐向室间沟心肌扩展,发育为房室传导系的希氏束、左右束支及心室腔面的小梁心肌.在心房,DES表达首先出现于C11期心房背侧壁,在C13期,可见静脉窦左背侧壁α-SCA、α-SMA、DES阳性心肌带与左心房底部、房室管背侧壁相延续,这条心肌带可能参与了人胚心静脉窦至房室管传导系的发育.C14～C16期,DES强阳性染色从窦房结经左、右静脉瓣及心房的背、腹侧壁延伸至房室管右侧壁,可能是原始的心房传导通路. 结论 心包腔尾端原始横膈间充质是人胚静脉窦心肌发生区,原始横膈间充质细胞逐渐分化为心肌细胞,添加到人胚心管静脉端,形成心静脉窦心肌.人胚心传导系心肌的分化始于房室管,随心管发育逐渐向动、静脉端扩展,在C16期,已分化为形态清晰可辨的DES阳性胚胎心传导系.     

超极化激活环核苷酸门控阳离子通道4、连接蛋白43和平足蛋白在小鼠胚胎心的表达与心传导系的发生

目的 探讨小鼠胚胎心传导系的发生机制。方法 用抗心肌肌球蛋白重链（MHC）、抗超极化激活环核苷酸门控阳离子通道4（HCN4）、抗缝隙连接蛋白43（CX43）和抗平足蛋白（podoplanin）抗体,对40只胚龄9~16d小鼠胚胎心进行连续石蜡切片并免疫组织化学或免疫荧光染色。结果 胚龄9d,较强的HCN4阳性表达集中在MHC阴性的静脉窦壁,随心脏发育,HCN4较强阳性表达逐渐向窦房结转移。胚龄11d开始,CX43阴性表达显示部位特异性。CX43阴性染色经窦房结沿右心房背侧壁和左、右静脉瓣向房室管背侧壁延伸。胚龄13d,左、右静脉瓣与房间隔底部融合后,进一步延续为房室管背侧壁发育中的CX43阴性染色的房室结,继而与室间隔顶部CX43阴性的房室束相连。胚龄9~10d,在MHC阳性心肌、心包腔背侧壁脏壁中胚层心肌前体细胞及静脉窦周间充质均显示podoplanin阳性表达。胚龄11~13d,podoplanin阳性间充质细胞沿心脏外表面扩展形成podoplanin阳性间皮样心外膜。结论 心脏发育早期,主起搏点位于静脉窦壁,起搏电位的产生早于收缩功能的发生。CX43阴性心肌是发育中的心传导系心肌,在胚龄11d即可观察到心传导系早期雏形。podoplanin参与促进心肌前体细胞向心肌细胞的分化。     

Strong desmin immunoreactivity in the myocardial sleeves around pulmonary veins,superior caval vein and coronary sinus supports the presumed arrhythmogenicity of these regions

Myocardial sleeve around human pulmonary veins plays a critical role in the pathomechanism of atrial fibrillation. Besides the well-known arrhythmogenicity of these veins, there is evidence that myocardial extensions into caval veins and coronary sinus may exhibit similar features. However, studies investigating histologic properties of these structures are limited. We aimed to investigate the immunoreactivity of myocardial sleeves for intermediate filament desmin, which was reported to be more abundant in Purkinje fibers than in ventricular working cardiomyocytes. Sections of 16 human (15 adult and 1 fetal) hearts were investigated. Specimens of atrial and ventricular myocardium, sinoatrial and atrioventricular nodes, pulmonary veins, superior caval vein and coronary sinus were stained with anti-desmin monoclonal antibody. Intensity of desmin immunoreactivity in different areas was quantified by the ImageJ program. Strong desmin labeling was detected at the pacemaker and conduction system as well as in the myocardial sleeves around pulmonary veins, superior caval vein, and coronary sinus of adult hearts irrespective of sex, age, and medical history. In the fetal heart, prominent desmin labeling was observed at the sinoatrial nodal region and in the myocardial extensions around the superior caval vein. Contrarily, atrial and ventricular working myocardium exhibited low desmin immunoreactivity in both adults and fetuses. These differences were confirmed by immunohistochemical quantitative analysis. In conclusion, this study indicates that desmin is abundant in the conduction system and venous myocardial sleeves of human hearts.     

HNK-1 in Morphological Study of Development of the Cardiac Conduction System in Selected Groups of Sauropsida

Human natural killer (HNK)-1 antibody is an established marker of developing cardiac conduction system (CCS) in birds and mammals. In our search for the evolutionary origin of the CCS, we tested this antibody in a variety of sauropsid species (Crocodylus niloticus, Varanus indicus, Pogona vitticeps, Pantherophis guttatus, Eublepharis macularius, Gallus gallus, and Coturnix japonica). Hearts of different species were collected at various stages of embryonic development and studied to map immunoreactivity in cardiac tissues. We performed detection on alternating serial paraffin sections using immunohistochemistry for smooth muscle actin or sarcomeric actin as myocardial markers, and HNK-1 to visualize overall staining pattern and then positivity in specific myocyte populations. We observed HNK-1 expression of various intensity distributed in the extracellular matrix and mesenchymal cell surface of cardiac cushions in most of the examined hearts. Strong staining was found in the cardiac nerve fibers and ganglia in all species. The myocardium of the sinus venosus and the atrioventricular canal exhibited transitory patterns of expression. In the Pogona and Crocodylus hearts, as well as in the Gallus and Coturnix ones, additional expression was detected in a subset of myocytes of the (inter)ventricular septum. These results support the use of HNK-1 as a conserved marker of the CCS and suggest that there is a rudimentary CCS present in developing reptilian hearts. Anat Rec, 302:69–82, 2019. © 2018 Wiley Periodicals, Inc.     

Topography of 3H-DHA, 3H-QNB, 3H-Dopamine, and 3H-DAGO Binding Sites Distribution in the Central Part of the Sinoatrial Node in Rat Heart

The topography of distribution of ³H-dihydroalprenolol, ³H-quinucledinyl benzilate, ³H-dopamine, and ³H-DAGO binding sites in the central part of the sinoatrial node in rat heart was studied by autoradiography after electrophysiological identification of the dominant pacemaker region location. Receptor asymmetry between the lateral and median regions of the central part of the sinoatrial node was shown. The dominant pacemaker region lay in the lateral area of the sinoatrial node; the number of binding sites for all four ligands was minimum in it. The number of binding sites gradually increased in the cranial and caudal directions from the dominant pacemaker region along the sinoatrial node artery (more smoothly in the caudal direction). The relative densities of bindings sites for ³H-dihydroalprenolol and ³H-dopamine were higher in the lateral region compared to the perinodal working myocardium, while the densities for ³H-quinucledinyl benzilate and ³H-DAGO were virtually the same. The distribution of binding sites along the artery in the median region of the sinoatrial node was even for ³H-quinucledinyl benzilate and ³H-DAGO. For ³H-DAGO these parameters were close to those in the perinodal atrial myocardium, for ³H-quinucledinyl benzilate somewhat lower. Curves presenting the distribution of binding site densities for ³H-dihydroalprenolol and ³H-dopamine in the median region of the sinoatrial node were similar, with a pronounced peak in the region contralateral to the dominant pacemaker region, and significantly higher binding parameters compared to those for the perinodal atrial myocardium. The difference consisted in higher density of ³H-dopamine binding sites in the median region of the sinoatrial node in comparison with the lateral region. Binding activity was maximum in the wall of the sinoatrial node artery. The distribution of binding sites for ligands to the main autonomic nervous system neurotransmitters in the rat heart sinoatrial node is heterogeneous. __________ Translated from Byulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 140, No. 10, pp. 472–477, October, 2005     

The nature of the superior sinus venosus defect

The location, and morphology, of the superior sinus venosus interatrial communication remains contentious. As part of a clinical study, we examined anatomic specimens and echocardiograms so as to clarify the arrangement of the normal atrial septal structures, and compared them with the arrangement found in the superior sinus venosus defect. The pathognomonic diagnostic criterion in the abnormal hearts was overriding of the intact muscular rim of the oval fossa by the mouth of the superior caval vein. This muscular rim is, in reality, a tube of myocardium which encloses a core of extracardiac adipose tissue. Understanding of this anatomic conundrum clarifies the understanding of the structures of both the normal atrial septum and sinus venosus defects. Clin. Anat. 11:349–352, 1998. © 1998 Wiley-Liss, Inc.     

The early development of the epicardium in Tupaia belangeri

Summary Development of the epicardium was studied in embryos of Tupaia belangeri from the 13th to 15th day of ontogeny. The greater part of the epithelium of the epicardium does not differentiate locally from the myoepicardium (cardiac splanchnopleure, splanchnic mesoderm), but rather from the coelomic epithelium of the septum transversum. The myoepicardium of the future atria and ventricles differentiates into myocardial cells only. On ontogenetic day 13, bulbar protrusions (the villi of Kurkiewicz 1909) are formed on the surface of the septum transversum and extend into the pericardial cavity, primarily between the sinoatrial and the ventricular regions of the embryonic heart. These protrusions are covered by flattened interdigitating cells, and they are filled with intercellular fluid of the mesenchyme of the septum transversum. Many mitoses are found among the cells. From these protrusions free vesicles are formed which are discharged into the pericardial cavity. The vesicles attach to the surface of the myoepicardium, i.e. to the developing myocardial cells. The vesicles open, and their cells spread out onto the surface of the heart to form the primary epicardium. This process begins on the dorsal surface of the heart, close to the protrusions of the septum transversum, there are, however, further isolated patches of primary epicardium in other regions of the surface of the heart. After the epicardial cells have settled onto the myocardium, mitoses become rare among them. On day 15, most of the myocardium is coated by the primary epicardium and the protrusions on the septum transversum disappear. A bare myocardium, as found on ontogenetic days 12 and 13 in Tupaia, might be a primitive (plesiomorphic) condition among chordates. In adult Branchiostoma, the coelomic epithelium which coats the contractile blood vessels had been found to differentiate into muscle cells that remain uncoated on the side facing the coelomic cavity (Franz 1933; Joseph 1914, 1928).     

Evolution and Development of the Atrial Septum

The complete division of the atrial cavity by a septum, resulting in a left and right atrium, is found in many amphibians and all amniotes (reptiles, birds, and mammals). Surprisingly, it is only in eutherian, or placental, mammals that full atrial septation necessitates addition from a second septum. The high incidence of incomplete closure of the atrial septum in human, so-called probe patency, suggests this manner of closure is inefficient. We review the evolution and development of the atrial septum to understand the peculiar means of forming the atrial septum in eutherian mammals. The most primitive atrial septum is found in lungfishes and comprises a myocardial component with a mesenchymal cap on its leading edge, reminiscent to the primary atrial septum of embryonic mammals before closure of the primary foramen. In reptiles, birds, and mammals, the primary foramen is closed by the mesenchymal tissues of the atrioventricular cushions, the dorsal mesenchymal protrusion, and the mesenchymal cap. These tissues are also found in lungfishes. The closure of the primary foramen is preceded by the development of secondary perforations in the septal myocardium. In all amniotes, with the exception of eutherian mammals, the secondary perforations do not coalesce to a secondary foramen. Instead, the secondary perforations persist and are sealed by myocardial and endocardial growth after birth or hatching. We suggest that the error-prone secondary foramen allows large volumes of oxygen-rich blood to reach the cardiac left side, needed to sustain the growth of the extraordinary large offspring that characterizes eutherian mammals. Anat Rec, 302:32–48, 2019. © 2018 The Authors. The Anatomical Record published by Wiley Periodicals, Inc. on behalf of American Association of Anatomists.     

Convergent evolutionary reduction of atrial septation in lungless salamanders

Nearly two thirds of the approximately 700 species of living salamanders are lungless. These species respire entirely through the skin and buccopharyngeal mucosa. Lung loss dramatically impacts the configuration of the circulatory system but the effects of evolutionary lung loss on cardiac morphology have long been controversial. For example, there is presumably little need for an atrial septum in lungless salamanders due to the absence of pulmonary veins and the presence of a single source of mixed blood flowing into the heart, but whether lungless salamanders possess an atrial septum and whether the sinoatrial aperture is located in the left or right atrium are unresolved; authors have stated opposing claims since the late 1800s. Here, we use micro‐computed tomography (μ‐CT) imaging, gross dissection and histological reconstruction to compare cardiac morphology among lungless plethodontid salamanders (Plethodontidae), salamanders with lungs, and the convergently lungless species Onychodactylus japonicus (Hynobiidae). Plethodontid salamanders have partial atrial septa and incomplete separation of the atrium into left and right halves. Partial septation is also seen in O. japonicus. Hence, lungless salamanders from two lineages convergently evolved similar morphology of the atrial septum. The partial septum in lungless salamanders can make it appear that the sinoatrial aperture is in the left atrium, but this interpretation is incorrect. Outgroup comparisons demonstrate that the aperture is located in a posterodorsal extension of the right atrium into the left side of the heart. Independent evolutionary losses of the atrial septum may have a similar developmental basis. In mammals, the lungs induce formation of the atrial septum by secreting morphogens to neighboring mesenchyme. We hypothesize that the lungs induce atrial septum development in amphibians in a similar fashion to mammals, and that atrial septum reduction in lungless salamanders is a direct result of lunglessness.     

Expression of Id2 in the second heart field and cardiac defects in Id2 knock-out mice

The inhibitor of differentiation Id2 is expressed in mesoderm of the second heart field, which contributes myocardial and mesenchymal cells to the primary heart tube. The role of Id2 in cardiac development is insufficiently known. Heart development was studied in sequential developmental stages in Id2 wildtype and knockout mouse embryos. Expression patterns of Id2, MLC-2a, Nkx2.5, HCN4, and WT-1 were analyzed. Id2 is expressed in myocardial progenitor cells at the inflow and outflow tract, in the endocardial and epicardial lineage, and in neural crest cells. Id2 knockout embryos show severe cardiac defects including abnormal orientation of systemic and pulmonary drainage, abnormal myocardialization of systemic and pulmonary veins, hypoplasia of the sinoatrial node, large interatrial communications, ventricular septal defects, double outlet right ventricle, and myocardial hypoplasia. Our results indicate a role for Id2 in the second heart field contribution at both the arterial and the venous poles of the heart.     

Early morphogenesis of the sinuatrial region of the chick heart: A contribution to the understanding of the pathogenesis of direct pulmonary venous connections to the right atrium and atrial septal defects in hearts with right isomerism of the atrial appendages

The morphogenesis of the sinuatrial region of embryonic hearts is still not well understood. Current matters of dispute are the topogenesis of the future pulmonary vein orifice and the topogenesis of the primary atrial septum. We analyzed the development of the sinuatrial region in chick embryos ranging from Hamburger and Hamilton (HH) stage 14 to 25. Our study disclosed three features of sinuatrial development. First, the primitive atrium of the HH stage 16 chick embryo heart has a separate inflow component. This inflow component takes up the mouth of the confluence of the systemic veins (sinus venosus) as well as the future mouth of the common pulmonary vein (pulmonary pit). The left portion of the atrial inflow component becomes incorporated into the left atrium and its right portion becomes incorporated into the right atrium. Rightward growth of the sinuatrial fold separates the sinus venosus from the left atrium. Second, the pulmonary pit originally forms as a bilaterally paired structure. Its left and right portions are connected to the left and right portions of the atrial inflow component, respectively. Normally, only the left portion of the pulmonary pit deepens to form the common pulmonary vein orifice, whereas the right portion disappears. Third, the primary atrial septum of the chick heart is not formed at the original midline of the embryonic heart, but is formed to the left of the original midline. This finding is in accord with molecular data suggesting that the primary atrial septum derives from the left heart‐forming field. Our findings shed new light on the pathogenesis of direct pulmonary venous connections to the right atrium and atrial septal defects in hearts with right isomerism of the atrial appendages. Anat Rec 290:168–180, 2007. © 2007 Wiley‐Liss, Inc.     

Atrioventricular node and input pathways: a correlated gross anatomical and histological study of the canine atrioventricular junctional region

To determine the architecture of the atrioventricular (AV) junctional region, structures in atrial preparations were correlated to those in serial sections made either parallel or perpendicular to the long axis of the AV node (AVN)/AV bundle complex. The results demonstrated the following for the first time: 1) A right medial atrial wall (MAW) extends anteriorly from the interatrial septum, superior to the interventricular septum (IVS). 2) An atrial interventricular septum (A-IVS) groove is located between the base of the MAW and the crest of the IVS. 3) Three atrionodal bundles converge to form a proximal AV bundle (PAVB), which in turn is contiguous with the AVN. The atrionodal bundles are associated with the MAW or the superomedial and inferolateral margins of the coronory sinus. Terminal portions of the atrionodal bundles and the PAVB reside within the A-IVS groove. The AV bundle was termed distal (DAVB) to avoid confusion. 4) The location of the AVN/DAVB complex topographically is deep to the apex of the septal cusp of the tricuspid valve subjacent to the MAW. Intracardially, the AVN/DAVB complex is within the central fibrous body. Significantly, this study resulted in the first unequivocal demonstration of discrete bundles of myocardial fibers associated with the atrial end of the AV node. Moreover, it appears likely that the atrionodal AV bundles are continuous with the sinoatrial nodal extensions, thereby forming internodal tracts.     

Atrial development in the human heart: an immunohistochemical study with emphasis on the role of mesenchymal tissues

The development of the atrial chambers in the human heart was investigated immunohistochemically using a set of previously described antibodies. This set included the monoclonal antibody 249-9G9, which enabled us to discriminate the endocardial cushion-derived mesenchymal tissues from those derived from extracardiac splanchnic mesoderm, and a monoclonal antibody recognizing the B isoform of creatine kinase, which allowed us to distinguish the right atrial myocardium from the left. The expression patterns obtained with these antibodies, combined with additional histological information derived from the serial sections, permitted us to describe in detail the morphogenetic events involved in the development of the primary atrial septum (septum primum) and the pulmonary vein in human embryos from Carnegie stage 14 onward. The level of expression of creatine kinase B (CK-B) was found to be consistently higher in the left atrial myocardium than in the right, with a sharp boundary between high and low expression located between the primary septum and the left venous valve indicating that the primary septum is part of the left atrial gene-expression domain. This expression pattern of CK-B is reminiscent of that of the homeobox gene Pitx2, which has recently been shown to be important for atrial septation in the mouse. This study also demonstrates a poorly appreciated role of the dorsal mesocardium in cardiac development. From the earliest stage investigated onward, the mesenchyme of the dorsal mesocardium protrudes into the dorsal wall of the primary atrial segment. This dorsal mesenchymal protrusion is continuous with a mesenchymal cap on the leading edge of the primary atrial septum. Neither the mesenchymal tissues of the dorsal protrusion nor the mesenchymal cap on the edge of the primary septum expressed the endocardial tissue antigen recognized by 249-9G9 at any of the stages investigated. The developing pulmonary vein uses the dorsal mesocardium as a conduit to reach the primary atrial segment. Initially, the pulmonary pit, which will becomes the portal of entry for the pulmonary vein, is located along the midline, flanked by two myocardial ridges. As development progresses, tissue remodeling results in the incorporation of the portal of entry of the pulmonary vein in left atrial myocardium, which is recognized because of its high level of creatine. Closure of the primary atrial foramen by the primary atrial septum occurs as a consequence of the fusion of these mesenchymal structures.     

Ultrastructural study of the sinus venosus in embryos of the dogfish (Scyliorhinus canicula)

An ultrastructural study of the development of the sinus venosus has been carried out on seven embryos of the dogfish (Scyliorhinus canicula L.) between 10.5 and 69 mm of total length (TL). The sinus venosus appears at the end of the looping process of the cardiac tube, namely in the 10.5 mm embryo, when the heart reaches its adult tetracameral S-form. The endocardium of the smallest embryo is constituted of a single layer of cubic cells. In larger embryo, these cells progressively acquire a squamous appearance as well as electron-dense cytoplasmic inclusions. The subendocardium is progressively populated by ganglion cells, Schwann cells and bundles of amyelinic fibers that can first be recognised in the embryo of 34 mm TL. Some subendocardial mesenchymal cells located in earlier embryos close to the entrance of the ducts of Cuvier might be their ectomesenchymal progenitors. The myocardium is initially constituted of a single layer of cubic cells. In the embryos of 19, 27 and 34 mm TL, the myocardium becomes multilayered, and the myocardiocytes develop myofibrils randomly arranged throughout the sarcoplasm. In later embryos, the myocardiocytes are innervated and arranged in oval bundles surrounded by a basal lamina. The epicardium covers the sinus venosus by the retrograde migration of the epithelium already established around the atrioventricular groove and, in a lesser degree, by the adhesion of mesothelial cells that are floating free in the pericardial cavity. This process has finished in the embryo of 34 mm TL. The differentiation of the sinus venosus (including the endocardial and myocardial differentiation as well as the epicardial covering) progresses in an anteroventral-posterodorsal direction.     

Distribution of <Superscript>3</Superscript>H-Dopamine and <Superscript>3</Superscript>H-DAGO Binding Sites in the Central Part of Rat Sinoatrial Node

Distribution of ³H-dopamine and ³H-DAGO binding sites was studied by autoradiography on semithin sections of total preparations of rat sinoatrial node. The relative density of ³H-dopamine and ³H-DAGO binding sites in the functional nucleus of the sinoatrial node was minimum and increased in the cranial and caudal directions. The level of ³H-dopamine binding in the perinodal atrial myocardium was appreciably lower (22±6%), while binding of ³H-DAGO was similar (76±16%) to that in the periarterial zone of the sinoatrial node. __________ Translated from Byulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 140, No. 8, pp. 210–214, August, 2005     

Development of the venous pole of the heart in the frog Xenopus laevis: A morphological study with special focus on the development of the venoatrial connections

The heart of lung‐breathing vertebrates normally shows an asymmetric arrangement of its venoatrial connections along the left‐right (L‐R) body axis. The systemic venous tributaries empty into the right atrium while the pulmonary venous tributaries empty into the left atrium. The ways by which this asymmetry evolves from the originally symmetrically arranged embryonic venous heart pole are poorly defined. Here we document the development of the venous heart pole in Xenopus laevis (stages 40–46). We show that, prior to the appearance of the mouth of the common pulmonary vein (MCPV), the systemic venous tributaries empty into a bilaterally symmetric chamber (sinus venosus) that is demarcated from the developing atriums by a circular ridge of tissue (sinu‐atrial ridge). A solitary MCPV appears during stage 41. From the time point of its first appearance onwards, the MCPV lies cranial to the sinu‐atrial ridge and to the left of the developing interatrial septum and body midline. L‐R lineage analysis shows that the interatrial septum and MCPV both derive from the left body half. The CPV, therefore, opens from the beginning into the future left atrium. The definitive venoatrial connections are established by the formation of a septal complex that divides the lumen of the venous heart pole into systemic and pulmonary venous flow pathways. This complex arises from the anlage of the interatrial septum and the left half of the sinu‐atrial ridge. Developmental Dynamics 240:1518–1527, 2011. © 2011 Wiley‐Liss, Inc.     

Isomyosin expression in developing chicken atria: a marker for the development of conductive tissue?

Summary Isomyosin expression patterns in embryonic chicken atria during the first two weeks of development were analyzed immunohistochemically.In the 3-days embryonic chicken heart (HH 19–20), strong coexpression of both isomyosins can be found as band-like zones at the lateral sides of the sinoatrial junction. The zones converge on the bottom of the atrium and continue as a band around the atrioventricular canal.In the 5-days heart (HH 27–28) the coexpression area encompasses the entire sinoatrial junction and extends into parts of the sinus venosus and into the dorsocaudal atrial wall.In the 7-days heart (HH 32–33) the relative extension of coexpression areas reaches its maximum. Coexpression is also found in a ring-like band in the ventral (bottom) wall of the atria peripheral to the ring-like band in the atrioventricular junction. The latter band has now become continuous with the coexpression area in the bottom of the interatrial septum. Caudally coexpression extends behind the atrioventricular cushions towards the interventricular septum and cranially coexpression of the atrioventricular junction has become continuus with that of the ring around the ouflow tract (cf Sanders et al.1986). In the second week of incubation a decrease of coexpression is observed.The isomyosin expression pattern described in this study has put forward additional arguments that the conductive tissue originates from areas that continue to express both isomyosins relatively late in development.