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First published online March 9, 2006
doi: 10.1242/10.1242/dev.02292

Development 133, 1399-1410 (2006)
Published by The Company of Biologists 2006
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Left-right lineage analysis of the embryonic Xenopus heart reveals a novel framework linking congenital cardiac defects and laterality disease

Ann F. Ramsdell1,2,3,*, Jayne M. Bernanke3 and Thomas C. Trusk3

1 Department of Cell and Developmental Biology and Anatomy, School of Medicine, University of South Carolina, Columbia, SC 29208, USA.
2 Program in Women's Studies, College of Arts and Sciences, University of South Carolina, Columbia, SC 29208, USA.
3 Department of Cell Biology and Anatomy and Cardiovascular Developmental Biology Center, Medical University of South Carolina, Charleston, SC 29425, USA.


Figure 1
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  		Fig. 1. Left-right lineage labeled embryo. Left and right cell lineages were microinjected with Oregon Green-conjugated dextran (green) and Alexa 647-conjugated dextran (red), respectively. Embryos were fixed and confocal images of whole mounts were collected after labeled embryos reached stages 45-46 (~5 days). A dorsal view of a labeled embryo is shown.

 

Figure 2
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  		Fig. 2. Left-right lineage composition in the normal heart. Left and right cell lineages in frontal tissue sections shown in this and the following figures were labeled by Oregon Green-conjugated dextran (green) and Alexa 647-conjugated dextran (red), respectively. (A) The right atrium (RA), left atrium (LA), interatrial septum (IAS), right cardinal vein (RCV) and left cardinal vein (LCV) are shown. (B) The atrioventricular canal (AVC) is shown. (C) The right aortic arch (RAA), left aortic arch (LAA), outflow tract (OFT) and ventricle (VEN) are shown. Green- and red-labeled blood cells are in the atrial chambers (A). Scale bar: 100 µm.

 

Figure 3
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  		Fig. 3. Left-right lineage composition in hearts of situs inversus embryos. Two different hearts (A-C, D-G) from situs inversus embryos showing the morphological left atrium (MLA), morphological right atrium (MRA), interatrial septum (IAS), right cardinal vein (RCV), left cardinal vein (LCV), right aortic arch (RAA), left aortic arch (LAA), atrioventricular canal (AVC), ventricle (VEN) and outflow tract (OFT). The anatomical positions of the left and right atria plus looping orientation are inverted in these hearts, and labeled blood cells are in the atrial chambers (A,D,G). Scale bar: 100 µm.

 

Figure 4
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  		Fig. 4. Left-right lineage composition in d-looped hearts of heterotaxic embryos. Three different hearts (A-C, D-G, H-K) show the right cardinal vein (RCV), left cardinal vein (LCV), right aortic arch (RAA), left aortic arch (LAA), atrioventricular canal (AVC), ventricle (VEN) and outflow tract (OFT). All of the hearts in this category contained an unseptated common atrium (CA). Scale bar: 100 µm.

 

Figure 5
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  		Fig. 5. Left-right lineage composition in l-looped hearts of heterotaxic embryos. Three different hearts (A-D, E-I, J-L) show the right cardinal vein (RCV), left cardinal vein (LCV), right aortic arch (RAA), left aortic arch (LAA), atrioventricular canal (AVC), ventricle (VEN) and outflow tract (OFT). The heart in A-D contains a common atrium (CA) and the heart in J-L has inverted atria, as indicated by the position of the morphological left atrium (MLA) and morphological right atrium (MRA), which are separated by an interatrial septum (IAS). Scale bar: 100 µm.

 

Figure 6
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  		Fig. 6. Left-right lineage composition in hearts of experimental situs solitus embryos with right-side CA-ALK4 expression. Three different hearts (A-C, D-F, G-I) are shown. The right cardinal vein (RCV), left cardinal vein (LCV), right aortic arch (RAA), left aortic arch (LAA), atrioventricular canal (AVC), ventricle (VEN), and outflow tract (OFT) are indicated. The heart in A-C contains a normally developed left atrium (LA) and right atrium (RA) separated by an interatrial septum (IAS). The heart in D-F contains a common atrium (CA) and the heart in G-I has inverted atria, as indicated by the position of the morphological left atrium (MLA) and morphological right atrium (MRA), which are separated by an IAS. Scale bar: 100 µm.

 

Figure 7
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  		Fig. 7. Left-right lineage composition in hearts of experimental situs solitus embryos with left-side CA-ALK4 expression. Three different hearts (A-C, D-G, H-J) are shown. The right cardinal vein (RCV), left cardinal vein (LCV), right aortic arch (RAA), left aortic arch (LAA), atrioventricular canal (AVC), ventricle (VEN), outflow tract (OFT) and aortic sac (AS) are indicated. The hearts in A-C and H-J each contain an unseptated common atrium (CA). Scale bar: 100 µm.

 

Figure 8
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  		Fig. 8. Pitx2c expression in normal and laterality defective embryos. Control and CA-ALK4-injected embryos were processed for whole-mount in situ hybridization to detect Pitx2c expression (arrowheads). (A) Control embryo, left-side view. (B) Right-side injected embryo, left and right-side views. (C) Right-side injected embryo, right-side view. (D) Left-side injected embryo, left-side view.

 

Figure 9
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  		Fig. 9. Left-right lineage composition in hearts of embryos situs following left-side DN-ALK4 expression. Three different hearts (A-C, D-F, G-I) are shown. The right cardinal vein (RCV), left cardinal vein (LCV), right aortic arch (RAA), left aortic arch (LAA), atrioventricular canal (AVC), ventricle (VEN), outflow tract (OFT) and aortic sac (AS) are indicated. Scale bar: 100 µm.

 

Figure 10
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  		Fig. 10. LR lineage composition of the normal heart. A three-dimensional schematic heart was created based on reconstructed serial sections of several hearts in this study. The exterior was pseudo-painted green and red to indicate left and right-side cell lineages, respectively. The right cardinal vein (RCV), left cardinal vein (LCV), right aortic arch (RAA), left aortic arch (LAA), left atrium (LA), right atrium (RA), atrioventricular canal (AVC), ventricle (VEN), outflow tract (OFT) and aortic sac (AS) are indicated in dorsal (A) and ventral (B) views.

 

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© The Company of Biologists Ltd 2006