Analysis Tools
mortality/aging
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• homozygotes start dying at ~E11.5
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cardiovascular system
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• homozygotes show a complete lack of vascular morphogenesis within the yolk sac
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• at E10.0, mutant erythrocytes reside within enlarged endothelial sinuses rather than a discrete network of vitelline blood vessels
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• at E10.0, mutant hearts are of normal size but occupy ~32.6% of the total embryo volume vs 15.5% in wild-type embryos
• no pericardial effusions are observed despite absence of cardiac function
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• at E10.0, mutant hearts show normal trabecular formation and compaction but lack seeding of endocardial cushions within the atrioventricular canal
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• at E9.5, mutant ventricular cardiomyocytes lack a normal parallel alignment of thick and thin filaments and display myofibrillar disorganization, absence of Z-lines and T-tubules, and a sparse sarcoplasmic reticulum
• however, no necrosis or cellular degradation is observed
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• at E9.5, homozygotes can produce electrically stimulated Ca2+ transients in primitive ventricles, although these tend to decay rapidly
• notably, mutants show an unexpected ~20% rise in basal Ca2+ levels after electrical stimulation (~10 transients), suggesting a role in the rapid phase of Ca2+ removal within early embryo cardiomyocytes
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• at E10.0, homozygotes show absence of circulation, despite normal cardiac looping and chamber formation
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• at E9.0, homozygotes have undegone normal cardiac looping but lack a discernible heart beat
• at E9.5, homozygotes exhibit absence of rhythmic spontaneous Ca2+ transients in primitive ventricles, consistent with lack of a functional contractile apparatus
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growth/size/body
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• homozygotes are of normal size at E9.0 but appear extremely growth retarded at E10.0 and E11.0
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• by E11.0, homozygotes are extremely small relative to wild-type embryos
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embryo
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• homozygotes show a complete lack of vascular morphogenesis within the yolk sac
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• at E10.0, mutant erythrocytes reside within enlarged endothelial sinuses rather than a discrete network of vitelline blood vessels
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• at E10.0, TUNEL analysis indicates significantly elevated levels of apoptosis throughout the entire body of the mutant embryo and extraembryonic membranes, but not within the mutant heart
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• homozygotes are of normal size at E9.0 but appear extremely growth retarded at E10.0 and E11.0
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• by E11.0, homozygotes are extremely small relative to wild-type embryos
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• at E10.0, homozygotes display elevated levels of apoptosis within both the endothelial component of the yolk sac and within erythrocytes
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cellular
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• at E10.0, TUNEL analysis indicates significantly elevated levels of apoptosis throughout the entire body of the mutant embryo and extraembryonic membranes, but not within the mutant heart
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