Katherine E. Yutzey

Development of heart valve leaflets and supporting apparatus in chicken and mouse embryos.

Abnormalities in valvuloseptal development significantly contribute to congenital heart defects, yet the underlying causes are complex and poorly understood. Early cardiac regulatory genes are differentially expressed during valvuloseptal development, consistent with novel functions during heart chamber formation in chicken and mouse embryos. Distinct valve cell lineages were identified in the leaflets, chordae tendineae, and myotendinous junctions with the papillary muscles based on restricted expression of extracellular matrix molecules. Specific cell types within these structures demonstrate characteristics of chondrogenesis and tendon development, identified by scleraxis, type II collagen, and tenascin expression. In chicken embryos, valve remodeling and maturation accompanies a decrease in mitotic index indicated by reduced bromodeoxyuridine incorporation. Analysis of Tie2‐cre × ROSA26R mice demonstrates that mature valve structures, including the atrioventricular and outflow tract semilunar valve leaflets, chordae tendineae, and the fibrous continuity that connects the septal leaflets of mitral and tricuspid valves, arise from endothelial cells of the endocardial cushions. Together, these studies provide novel insights into the origins and cell lineage diversity of mature valve structures in the developing vertebrate heart. Developmental Dynamics 230:239–250, 2004.

NFATc3 and NFATc4 Are Required for Cardiac Development and Mitochondrial Function

Abstract— Activation of the nuclear factor of activated T-cell (NFAT) family of transcription factors is associated with changes in gene expression and myocyte function in adult cardiac and skeletal muscle. However, the role of NFATs in normal embryonic heart development is not well characterized. In this report, the function of NFATc3 and NFATc4 in embryonic heart development was examined in mice with targeted disruption of both nfatc3 and nfatc4 genes. The nfatc3−/−nfatc4−/− mice demonstrate embryonic lethality after embryonic day 10.5 and have thin ventricles, pericardial effusion, and a reduction in ventricular myocyte proliferation. Cardiac mitochondria are swollen with abnormal cristae, indicative of metabolic failure, but hallmarks of apoptosis are not evident. Furthermore, enzymatic activity of complex II and IV of the respiratory chain and mitochondrial oxidative activity are reduced in nfatc3−/−nfatc4−/− cardiomyocytes. Cardiac-specific expression of constitutively active NFATc4 in nfatc3−/−nfatc4−/− embryos prolongs embryonic viability to embryonic day 12 and preserves ventricular myocyte proliferation, compact zone density, and trabecular formation. The rescued embryos also maintain cardiac mitochondrial ultrastructure and complex II enzyme activity. Together, these data support the hypothesis that loss of NFAT activity in the heart results in a deficiency in mitochondrial energy metabolism required for cardiac morphogenesis and function.

Thyroid transcription factor‐1, hepatocyte nuclear factor‐3β and surfactant protein A and B in the developing chick lung

Expression of surfactant proteins SP‐A, SP‐B and the transcription factors TTF‐1 and HNF‐3β was identified by immunohistochemistry in the developing chicken. SP‐B, a small hydrophobic peptide critical for lung function and surfactant homeostasis in mammals, was detected in the epithelial cells of parabronchi in embryonic chicken lung from the 15th day of incubation, prior to the onset of the breathing movements and was expressed at high levels in the posthatching chicken lung. SP‐A, an abundant surfactant protein involved in innate defence of the mammalian lung, was detected in the chick embryo in subsets of epithelial cells in the mesobronchus, starting from d 15 and was detected in the posthatching chicken lung. The transcription factors hepatocyte nuclear factor 3β (HNF‐3β) and thyroid transcription factor‐1 (TTF‐1), both regulators epithelial cell differentiation and gene expression in mammalian species, were detected at the onset of lung bud formation (d 4 of incubation) and throughout lung development. Abundant nuclear expression was detected in nuclei of respiratory epithelial cells of developing bronchial tubules for both transcription factors. In contrast to the surfactant proteins, expression of both TTF‐1 and HNF‐3β decreased markedly in posthatching chicken lung. The expression of SP‐A and SP‐B in chick lung demonstrates the conservation of surfactant proteins in vertebrates. The temporospatial pattern of TTF‐1 and HNF‐3β overlaps with that of SP‐A and SP‐B, supporting their potential roles in chick lung development and demonstrating the conservation of regulatory mechanisms contributing to gene expression in respiratory epithelial cells in vertebrates.