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FIG. 3. Milestones of early cardiac development in Xenopus visualized by confocal images of double-immunostained embryos. Embryos selected to represent a chronology of stages were double labeled with anti-tropomyosin (CH1) (red) and anti-fibrillin (JB3) (green) and optically sectioned. For each stage, the set of optical sections (28 to 40 sections, 4 to 6 um thick) was digitally compiled in increments of 4 to 6 sections. Resulting optical thick sections that aid in the visualization of Xenopus heart development are presented. The order of presentation for stages with m ore than one image is ventral to dorsal. Image A is a parasagittal view of a stage 24 hemisectioned embryo. The stage 26 embryo in B is shown from the left lateral view. Images of stages 28 (C and D) and 31 (E) are ventral views of heart regions with the anterior of the em bryos placed at the top. At stage 24, the heart anlage is present but not immunohistochemically active. By stage 26, the first presence of antibody reactivity is noted near the ventral midline (ventral midline lies between arrowheads). At stage 28 (C and D), differentiation of the heart anlage is clear; the pericardial cavity can be discerned. Stage 31 (E)he heart region extends ventrally beyond the pericardial cavity and is slightly bent laterally to the embryo right; the myocardial wall does not yet close. Scale bars as denoted. |
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FIG. 1. Cardiac muscle development in Xenopus. Confocal images of the heart regions of six stages of embryos (27-46) are presented (ventral views), each adjacent to a diagrammatic representation of an embryo (Nieuwkoop and Faber, 1967) at the corresponding stage. Prior to microscopy, stage 27-33 embryos were immunolabeled with anti-tropomyosin (CH1) and stages 356 with anti-cardiac troponin T (CT3). The secondary antibody used for all stages was conjugated to Cy5. Each confocal image represents a digitally colored compilation of 30-53 optical sections taken (in gray scale) 5 um apart. All confocal images were taken at the same magnification and are presented with the anterior of the embryo at the top. Scale bar, 100 um |
