The final morphogenesis of the cardiac outflow tract (OFT) depends on coordinated cell-cell interactions among the heart tip, migrating heart-anchoring cells (HANCs) and the dorsally growing cardiac outflow muscles (COMs). This is a three-dimensional reconstruction of the cardiac outflow tract (horizontal rotation) from stage-16 wild-type embryos stained for Ladybird early (Lbe; blue), Msp300 (green), and Tinman (Tin; red). HANC cells (in blue) overlap dorsally the most anterior aorta. The COM muscles (in green) are attached on both sides to the most anterior Lbe-expressing cardioblasts (at this stage Lbe expression in the cardioblasts and pericardial cells is very week, and rendering Lbe signal visible resulted in an increased background). Note that ventrally to the tip of aorta are located cephalic vascular rudiment cells (cvr), which are Tin-positive, Msp300-negative. Movie is SI Movie 1A in Proc Natl Acad Sci USA (2008) 105: 2475-2480. Movies from this publication include CIL# 25536, 25537, 25538, 25539, 25540, 25541.
Stage 16 embryos were stained with primary antibodies: mouse anti-Lbe, rabbit anti-Tim, guinea pig anti-Msp300. Species-specific secondary antibodies were conjugated to Alexa Fluor 488, Cy3 or Cy5. The triple stained embryos were visualized using a Zeiss LSM 510 Meta confocal microscope. Images representing 0.5 mm optical sections were compiled and processed by Volocity software (Improvision).
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