To evaluate normal embryonic mouse heart development using Doppler echocardiography and to quantify changes in normal embryonic mouse cardiac function with increasing gestational age from the time of cardiac septation, a new method was applied using Doppler echocardiography. Trisomic embryos were screened to evaluate a model of abnormal cardiac anatomy. The development of the embryonic heart in mice has been well studied anatomically, but there are limited physiologic studies. A new method has been developed to assess the mouse fetal heart in a similar fashion to the current use of echocardiography in the chick embryo and the human fetus. This method was applied to normal mouse embryos known to survive and to abnormal trisomy embryos that die during gestation and have cardiac failure. To analyze early normal embryonic heart hemodynamics, Doppler echocardiograms were performed on n = 129 C57B1/6J mouse embryos from d 10 through 19 of gestation and 20 embryos with trisomy 16 (gestational d 11-14). The maximal blood velocities recorded at the inflow and outflow of the embryonic heart were analyzed for heart rate, peak early and peak late inflow and outflow velocities, and measurements were made of systolic ejection, filling, and other time intervals normalized for heart rate. A high velocity holosystolic or diastolic velocity with altered time intervals was identified as atrioventricular or semilunar valvular regurgitation, respectively. Inflow and outflow velocities increased with increasing gestational age. The time period of isovolemic contraction time was present before and undetectable after gestational d 17, whereas the total filling time increased. Ejection time and isovolemic relaxation time had no significant change. No valvular regurgitation was detected in normal embryos. These echocardiographic patterns are similar to those observed for human embryos. Abnormal Doppler findings were present (inflow or outflow valvular regurgitation) in 55% of trisomy 16 embryos. Echocardiographic data can now be obtained beginning at d 11 in the mouse embryo for analyses relating to abnormal heart development. A noninvasive technique may be invaluable to monitor the physiologic condition of embryos within a litter and to detect and monitor those embryos where heart defects may be expected. Qualitative markers of embryonic congestive heart failure such as valvular regurgitation may be present and detectable with structural valvular abnormalities or failing cardiac physiology. The mouse embryo is an appropriate animal model to analyze normal and abnormal mammalian heart development and function.

During early heart development the expression pattern of N-cadherin, a calcium-dependent cell adhesion molecule, suggests its involvement in morphoregulation and the stabilization of cardiomyocyte differentiation. N-cadherin's adhesive activity is dependent upon its interaction with the intracellular catenins. An association with alpha-catenin and beta-catenin also is believed to be involved in cell signaling. This study details the expression patterns of alpha-catenin, beta-catenin, and gamma-catenin, during definition of the cardiac cell population as distinct compartments in the anterior regions of the chick embryo between stages 5 and 9. The restriction of N-cadherin/catenin localization at stage 5+ from a uniform pattern in vivo, to specific cell clusters that demarcate areas where mesoderm separation is initiated, suggests that the N-cadherin/catenin complex is involved in boundary formation and in the subsequent cell sorting. The latter two processes lead to the specification and formation of the somatic and cardiac splanchnic mesoderm. N-cadherin colocalized with alpha- and beta-catenin at the cell membrane before and during the time that its expression becomes restricted to the lateral mesoderm and continues cephalocaudad into stage 8. These proteins continue to colocalize in the myocardium of the tubular heart. Plakoglobin is not expressed in this region during stages 6-8, but is detected in the myocardium later at stage 13. The observed in vivo expression patterns of alpha-catenin, beta-catenin, and plakoglobin suggest that these proteins are directly linked with the developmental regulation of cell junctions, as cardiac cells become stably committed and phenotypically differentiated to eventually form a mature myocardium. The localization of N-CAM also was analyzed during these stages to determine whether the N-cadherin-catenin localization was unique or whether other cell adhesion molecules were expressed similarly. The results indicate that the unique pattern of N-cadherin expression is not shared with N-CAM. We also show that perturbation of N-cadherin using a function perturbing N-cadherin antibody (NCD-2) inhibits normal early heart development and myogenesis in a cephalocaudad, stage-dependent manner. We propose a model whereby myocardial cell compartmentalization also defines the endocardial population. The presence of beta-catenin suggests that a similar signaling pathway involving Wnt (wingless)-mediated events may function in myocardial cell compartmentalization during early vertebrate heart development, as in Drosophila contractile vessel development.

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Fetal cardiac wall thickness, cardiac systolic and diastolic functions are affected by GDM independently of glycemic control.