Michael Choy

Fate of the atrioventricular endocardial cushions in the developing chick heart.

ABSTRACT: To determine the fate of the atrioventricular endocardial cushions in cardiac development, we used staining methods for extracellular fibronectin, which is abundant in the endocardial cushions, and actin, which is abundant in the myocytes. White Leghorn chick embryo hearts were harvested at Hamburger and Hamilton stages 26 to 36, and serial sections of the atrioventricular valve region were stained. Before atrioventricular valve formation, fibronectin and actin staining reveal separation between the fibronectin-rich endocardial cushions and the actin-rich myocardial layer. The developing mitral valve leafets at all of the observed stages contain a fibronectinrich matrix but no actin-rich myocytes. In contrast, the tricuspid band includes both fibronectin matrix and actinrich cells. We conclude that the mitral valve leaflets in the chick form predominantly from the endocardial cushion tissue, and the tricuspid band receives contributions from both the endocardial cushions and surrounding myocardium.

Regulation of proliferation of embryonic heart mesenchyme: Role of transforming growth factor-β1 and the interstitial matrix

Proliferation of atrioventricular cushion mesenchyme of the embryonic avian heart maintained in three-dimensional aggregate culture is stimulated by interaction with the interstitial matrix. Chicken serum or transforming growth factor-beta 1, which stimulates proliferation, induces matrix deposition in regions of the aggregate showing high labeling indices with tritiated thymidine. Dispersed heart mesenchyme interstitial matrix introduced into serum-free culture is incorporated into the aggregate and stimulates cellular proliferation similar to serum or transforming growth factor-beta 1. Proliferation is reversibly inhibited by the peptide Gly-Arg-Gly-Asp-Ser-Pro. It is suggested that transforming growth factor-beta 1 stimulates the production of interstitial matrix and that a sufficient stimulus for proliferation in this system is the presence of the matrix, which acts as the adhesive support for cellular anchorage.

Fibroblast growth factor-2 stimulates embryonic cardiac mesenchymal cell proliferation

The proliferation response of stage 36 chick atrioventricular valve mesenchymal cells to fibroblast growth factor‐2 (FGF‐2) was studied in the tissue‐like environment of three‐dimensional cell aggregates maintained in organ culture. The mitogenic effects of FGF‐2 on mesenchymal tissue depended on the FGF‐2‐stimulated formation of a fibronectin‐containing extracellular matrix. The matrix was absent in unstimulated aggregates, and co‐localized with regions of actively proliferating cells in stimulated aggregates. Inhibition of fibronectin matrix formation by the inclusion of Arg‐Gly‐Asp‐containing peptides, which compete with fibronectin for binding to the cell surface α5β1 integrin receptors, abolished the proliferation effects of FGF‐2. Inhibition of sulfation of cell surface glycosaminoglycans by treatment with sodium chlorate significantly reduced both the formation of the fibronectin matrix and cell proliferation in response to FGF‐2, suggesting an involvement of the low‐affinity sulfated glycosaminoglycan FGF receptor system. Thus, the FGF‐stimulated growth of embryonic atrioventricular valve mesenchyme in vitro involves the production of a fibronectin matrix. We suggest that the stimulation of the fibronectin matrix represents an essential element in growth factor signaling of mesenchymal tissue, with the matrix serving as an anchorage substratum for the proliferating cells.

Absent Pulmonary Valve with Tricuspid Atresia or Severe Tricuspid Stenosis: Report of Three Cases and Review of the Literature

Absence of the pulmonary valve occurs usually in association with tetralogy of Fallot and occasionally with an atrial septal defect or as an isolated lesion. Very rarely it occurs with tricuspid atresia, intact ventricular septum, and dysplasia of the right ventricular free wall and of the ventricular septum. We present the clinical, anatomic, and histologic findings of a new case, and for the first time, the data from two patients with absent pulmonary valve and severe tricuspid stenosis, who exhibited similar histologic findings. We also reviewed the clinical and anatomic data of 24 previously published cases and compared them with the new cases. In all three new cases, the myocardium of the right ventricle was very abnormal. In the two cases with tricuspid stenosis, large segments of myocardium were replaced with sinusoids and fibrous tissue. In the case with tricuspid atresia, the right ventricular free wall contained only fibroelastic tissue. The ventricular septum in all three patients showed asymmetric hypertrophy and in two of the three patients, multiple sinusoids had replaced large segments of myocardial cells. The left ventricular free wall myocardium and the walls of the great arteries were unremarkable. Our data indicate that myocardial depletion involving the right ventricular free wall and the ventricular septum and its replacement by sinusoids and fibroelastic tissue occur not only in cases of absent pulmonary valve with tricuspid atresia but also in cases of absent pulmonary valve with tricuspid stenosis. The degree of myocardial depletion varies and is more severe when the tricuspid valve is atretic.

Transforming growth factor-β1 localized within the heart of the chick embryo

SummaryTransforming growth factor-β1 is a pleiotropic peptide mediator of growth, differentiation, and extra-cellular matrix synthesis. In the embryonic chick heart prior to the formation of the endocardial cushions, evidence from in vitro experiments suggests that transforming growth factor-β1 may be an inducer of the differentiation of atrioventricular endothelial cells into endocardial cushion mesenchyme. Further in vitro evidence suggests that the factor stimulates mesenchymal cell proliferation, and, thus, growth of the cushions. Using an antibody made against a peptide duplicating the aminoterminal 30 amino acid sequence of transforming growth factor-β1, we stained sections of stage 11, 18, 23, 26, and 36 chick hearts by an in situ immunofluorescence technique. Transforming growth factor-β1 staining localized to the endocardial surface and epicardial surface of the stage 11 heart, but it decreased from these locations in later stages. The cardiac jelly (stage 11), endocardial cushions (stage 18, 23, and 26), and, subsequently, the heart valve leaflets (stage 36) stained intensely for the growth factor.

Test Characteristics of a Level I or II Fetal Ultrasound in Detecting Structural Heart Disease

At the University of California Davis Medical Center, a screening fetal ultrasound examination (level I or II) incorporates a comprehensive segmental evaluation of the fetal heart. This study evaluated the reliability of the fetal ultrasound exam in the detection of abnormal heart anatomy. Our retrospective study reviewed results of 614 antenatal patients that had a screening fetal ultrasound exam. All patients subsequently underwent a detailed targeted fetal cardiac ultrasound exam performed by a pediatric cardiac sonographer and reviewed by a board-certified pediatric cardiologist. Of these 614 patients, 60 fetuses had structural heart disease by the targeted fetal exam. The screening fetal ultrasound exam correctly identified 55 of the 60, with 5 false negatives (8.3% false-negative rate) and 1 false positive (1.7% false-positive rate). Our study suggests that if a screening fetal ultrasound exam incorporates a segmental evaluation of the fetal heart it can reliably detect abnormal heart anatomy. At our institution a targeted fetal cardiac exam is now used to confirm and provide detailed assessment of the heart anatomy when a screening fetal exam is positive for heart disease.

Cardiomyopathy induced by phenylpropanolamine

A 14-year-old girl had evidence of an acute cardiomyopathy after a minimal overdose of phenylpropanolamine. She had myocardial dysfunction, ventricular dysrhythmia, and secondary pulmonary edema without any associated systemic hypertension. The cardiomyopathy resolved after several days.

Bisdiamine inhibits extracellular matrix formation and cell proliferation of atrioventricular mesenchyme from developing chick heart valves

Abnormalities of the cushion tissues lead to atrioventricular septal defects (AVSD) and truncus arteriosus (TA). Bisdiamine exposure in the embryo frequently causes AVSD and TA in the newborn chick, mouse, or rat. We studied the effects of bisdiamine on mesenchymal cells grown in aggregate culture isolated from the developing atrioventricular valves of the stage-36 chick embryo. Fibronectin extracellular matrix formation and cell proliferation in the aggregates were assessed in various media. Chick serum stimulated the cells to produce an extracellular matrix and to divide, and the inclusion of bisdiamine inhibited both responses. If we isolated an extracellular matrix from a monolayer of mesenchymal cells and added the sonicated matrix to the medium containing serum and bisdiamine, the matrix incorporated into the aggregates and the cells entered the mitotic cycle. Our previous work established that cells need to attach to an intact extracellular matrix to begin cell division. Thus, we suggest that bisdiamine inhibits the normal formation of the extracellular matrix, leading to reduced cell proliferation, but it does not affect matrix-cell interaction. The lack of cushion growth in situ may be the cause of AVSD or TA.