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Immunofluorescent <t>Tfap2α</t> and Nkx2.5 expression in the OFT area of the heart in wild-type and Lrp2 knockout embryos at E10.5 and E11 . 5. The mesenchymal Tfap2α (red)- and Nkx2.5 (green)-positive cells are separate populations in both the wild-type (WT) and knockout (KO) embryos. No co-expression was detected around the aortic sac (AoS) or the pharyngeal arch arteries. Note: a part of the ectodermal cells express Tfap2α, but are not involved in outflow tract (OFT) septation. The flow divider (FD) is a part of the Nkx2.5-positive cell population and is present in between the most caudal pharyngeal arch arteries (C,F). Boxed areas in B and E are shown in C and F, respectively. Scale bars: 50 µm.
Anti Tfap2α, supplied by GeneTex, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Immunofluorescent Tfap2α and Nkx2.5 expression in the OFT area of the heart in wild-type and Lrp2 knockout embryos at E10.5 and E11 . 5. The mesenchymal Tfap2α (red)- and Nkx2.5 (green)-positive cells are separate populations in both the wild-type (WT) and knockout (KO) embryos. No co-expression was detected around the aortic sac (AoS) or the pharyngeal arch arteries. Note: a part of the ectodermal cells express Tfap2α, but are not involved in outflow tract (OFT) septation. The flow divider (FD) is a part of the Nkx2.5-positive cell population and is present in between the most caudal pharyngeal arch arteries (C,F). Boxed areas in B and E are shown in C and F, respectively. Scale bars: 50 µm.

Journal: Disease Models & Mechanisms

Article Title: Common arterial trunk and ventricular non-compaction in Lrp2 knockout mice indicate a crucial role of LRP2 in cardiac development

doi: 10.1242/dmm.022053

Figure Lengend Snippet: Immunofluorescent Tfap2α and Nkx2.5 expression in the OFT area of the heart in wild-type and Lrp2 knockout embryos at E10.5 and E11 . 5. The mesenchymal Tfap2α (red)- and Nkx2.5 (green)-positive cells are separate populations in both the wild-type (WT) and knockout (KO) embryos. No co-expression was detected around the aortic sac (AoS) or the pharyngeal arch arteries. Note: a part of the ectodermal cells express Tfap2α, but are not involved in outflow tract (OFT) septation. The flow divider (FD) is a part of the Nkx2.5-positive cell population and is present in between the most caudal pharyngeal arch arteries (C,F). Boxed areas in B and E are shown in C and F, respectively. Scale bars: 50 µm.

Article Snippet: After deparaffination and rehydration of the slides, immunohistochemical staining was performed with the antibodies anti-atrial myosin light chain 2a (MLC-2a) as a myocardial marker (1/4000, kindly provided by S. W. Kubalak, Medical University of South Carolina, SC), anti-Nkx2.5 as a (pre)myocardial marker and a marker for SHF-derived cells (1/4000, Santa Cruz Biotechnology, Inc., CA, SC-8697), anti-LPR2 (1/800, kindly provided by J. Herz, University of Texas Southwestern Medical Center, TX), anti-WT1 as a marker for the epicardium and early migrating EPDCs (1/500, Calbiochem, ONCOCA1026), anti-Tfap2α as an NCC marker (1/500, GeneTex Inc., GTX62588) and anti-CD31 (M-20) as a marker for endothelium and endocardium (1/4000, Santa Cruz Biotechnology, Inc., CA, SC-1506).

Techniques: Expressing, Knock-Out

Outflow tract septation characteristics with 3D reconstructions and histological sections of E10.5 wild-type and knockout embryos. (A-D) In the reconstructions (WT, wild type; KO, knock out), the second heart field (SHF) has been depicted in yellow; the neural crest cells (NCCs) surrounding the aortic sac (AoS; in red) and the 4th (4; in red) and 6th (6; in blue) pharyngeal arch arteries have been removed. Only the NCC mesenchymal population (dark green) and the NCCs (bright green) that are embedded in the endocardial cushions (not shown) are depicted. The 6th pharyngeal arch arteries and the pulmonary arteries (PA) are depicted in blue. (A,E) Ventral view of a WT embryo: a population of SHF cells is seen to cross from dorsal to ventral, along the 4th pharyngeal arch artery on the right (aortic) side of the AoS up to the myocardial borderline of the AoS (asterisk). (B,I) Left lateral view. On the left (pulmonary) side of the AoS below the 6th pharyngeal arch artery, an SHF population is seen (black arrowhead). The section levels are indicated in B (upper section level: E,F and lower level: I,J). Not visible in the reconstructions is the Nkx2.5-positive SHF population in front of the gut (‘G’) that forms a ridge (FD, flow divider) in the dorsal wall of the AoS between the right and left 6th pharyngeal arch arteries (arrow in E) and negative for the NCC marker Tfap2α (arrow in F). NCCs are, however, abundantly present around the 6th pharyngeal arch arteries (brown Tfap2α cells in F,J). In the 3D reconstructions of the knockout embryo (C,D), the frontal SHF population is less well developed (C) and the population on the left (pulmonary) side remains in the midline dorsal position between the 6th pharyngeal arch arteries and does not properly extend anteriorly (D). The mesenchymal NCC population (dark green) on the left (pulmonary) side already reaches the NCCs in the endocardial OFT (D). The midline dorsal FD (arrows in G,H,K,L) protrudes prominently into the AoS and separates the 6th pharyngeal arch arteries. Scale bars: 50 µm. See also the online supplemental videos (E10.5_video_1080p.mpg) and interactive pdf reconstructions (E10.5_WT_revise.pdf and E10.5_KO_revise.pdf) at http://www.caskanatomy.info/research/supplement_megalin.html . S, superior; R, right; L, left; I, inferior; A, anterior; P, posterior; ec, endocardial cushions.

Journal: Disease Models & Mechanisms

Article Title: Common arterial trunk and ventricular non-compaction in Lrp2 knockout mice indicate a crucial role of LRP2 in cardiac development

doi: 10.1242/dmm.022053

Figure Lengend Snippet: Outflow tract septation characteristics with 3D reconstructions and histological sections of E10.5 wild-type and knockout embryos. (A-D) In the reconstructions (WT, wild type; KO, knock out), the second heart field (SHF) has been depicted in yellow; the neural crest cells (NCCs) surrounding the aortic sac (AoS; in red) and the 4th (4; in red) and 6th (6; in blue) pharyngeal arch arteries have been removed. Only the NCC mesenchymal population (dark green) and the NCCs (bright green) that are embedded in the endocardial cushions (not shown) are depicted. The 6th pharyngeal arch arteries and the pulmonary arteries (PA) are depicted in blue. (A,E) Ventral view of a WT embryo: a population of SHF cells is seen to cross from dorsal to ventral, along the 4th pharyngeal arch artery on the right (aortic) side of the AoS up to the myocardial borderline of the AoS (asterisk). (B,I) Left lateral view. On the left (pulmonary) side of the AoS below the 6th pharyngeal arch artery, an SHF population is seen (black arrowhead). The section levels are indicated in B (upper section level: E,F and lower level: I,J). Not visible in the reconstructions is the Nkx2.5-positive SHF population in front of the gut (‘G’) that forms a ridge (FD, flow divider) in the dorsal wall of the AoS between the right and left 6th pharyngeal arch arteries (arrow in E) and negative for the NCC marker Tfap2α (arrow in F). NCCs are, however, abundantly present around the 6th pharyngeal arch arteries (brown Tfap2α cells in F,J). In the 3D reconstructions of the knockout embryo (C,D), the frontal SHF population is less well developed (C) and the population on the left (pulmonary) side remains in the midline dorsal position between the 6th pharyngeal arch arteries and does not properly extend anteriorly (D). The mesenchymal NCC population (dark green) on the left (pulmonary) side already reaches the NCCs in the endocardial OFT (D). The midline dorsal FD (arrows in G,H,K,L) protrudes prominently into the AoS and separates the 6th pharyngeal arch arteries. Scale bars: 50 µm. See also the online supplemental videos (E10.5_video_1080p.mpg) and interactive pdf reconstructions (E10.5_WT_revise.pdf and E10.5_KO_revise.pdf) at http://www.caskanatomy.info/research/supplement_megalin.html . S, superior; R, right; L, left; I, inferior; A, anterior; P, posterior; ec, endocardial cushions.

Article Snippet: After deparaffination and rehydration of the slides, immunohistochemical staining was performed with the antibodies anti-atrial myosin light chain 2a (MLC-2a) as a myocardial marker (1/4000, kindly provided by S. W. Kubalak, Medical University of South Carolina, SC), anti-Nkx2.5 as a (pre)myocardial marker and a marker for SHF-derived cells (1/4000, Santa Cruz Biotechnology, Inc., CA, SC-8697), anti-LPR2 (1/800, kindly provided by J. Herz, University of Texas Southwestern Medical Center, TX), anti-WT1 as a marker for the epicardium and early migrating EPDCs (1/500, Calbiochem, ONCOCA1026), anti-Tfap2α as an NCC marker (1/500, GeneTex Inc., GTX62588) and anti-CD31 (M-20) as a marker for endothelium and endocardium (1/4000, Santa Cruz Biotechnology, Inc., CA, SC-1506).

Techniques: Knock-Out, Marker

Outflow tract septation characteristics with 3D reconstructions and histological sections of E11.5 wild-type and knockout embryos. Comparable to <xref ref-type=Fig. 4 , in the reconstructions (A-D) the second heart field (SHF) has been depicted in yellow; the neural crest cells (NCCs) surrounding the aorta (Ao; in red) and the 4th (4; in red) and 6th (6; in blue) pharyngeal arch arteries have been removed. Only the NCC mesenchymal population (dark green) and the NCCs (bright green) that are embedded in the endocardial cushions (not shown) are depicted. The 6th pharyngeal arch arteries and the pulmonary arteries (PA) are depicted in blue. The dorsal mid-sagittal SHF population has moved to the left and has extended a large band of cells to the left (pulmonary) side (P) of the OFT extending along the pulmonary vessel wall and the myocardial OFT (A,B,E, black arrowhead, F). The flow divider (FD) has separated the 6th pharyngeal arch arteries and has also moved to the left (E,F). The Tfap2α-positive NCCs (G) are encircled with a striped line. They have taken up a right-sided position next to the FD and are connected to the NCCs in the opposing endocardial cushions (ec). The NCC is forming the aorto-pulmonary septum (E-G, double-headed arrow), viewed in the 3D reconstruction (B) at the connection between dark and bright green. In the knockout embryo, both the SHF and the NCC population on the right (aortic) side are not well developed (C). Because the dorsal SHF population (not shown in the reconstruction) remains in midline (Nkx2.5-positive cells in sections H,I), there is hardly an extension along the left (pulmonary) side (C,D and H,I, black arrowhead). There is a marked Tfap2α (J) NCC population, however, on the left (pulmonary) side that connects to the bright green NCCs in the endocardial cushions. This does not lead to septation of the aortic sac, which now has become a common arterial trunk (CAT) (H-J). The FD is still prominently present and is Nkx2.5 (SHF)-positive (H,I, arrow) and Tfap2α (NCC)-negative (J; arrow). Section levels have been indicated in the reconstructions (A,C). Scale bars: 50 µm. See also the online supplemental videos (E11.5_video_1080p.mpg) and interactive pdf reconstructions (E11.5_WT_revise.pdf and E11.5_KO_revise.pdf) on http://www.caskanatomy.info/research/supplement_megalin.html . S, superior; R, right; L, left; I, inferior; A, anterior; P, posterior; Pt, pulmonary trunk. " width="100%" height="100%">

Journal: Disease Models & Mechanisms

Article Title: Common arterial trunk and ventricular non-compaction in Lrp2 knockout mice indicate a crucial role of LRP2 in cardiac development

doi: 10.1242/dmm.022053

Figure Lengend Snippet: Outflow tract septation characteristics with 3D reconstructions and histological sections of E11.5 wild-type and knockout embryos. Comparable to Fig. 4 , in the reconstructions (A-D) the second heart field (SHF) has been depicted in yellow; the neural crest cells (NCCs) surrounding the aorta (Ao; in red) and the 4th (4; in red) and 6th (6; in blue) pharyngeal arch arteries have been removed. Only the NCC mesenchymal population (dark green) and the NCCs (bright green) that are embedded in the endocardial cushions (not shown) are depicted. The 6th pharyngeal arch arteries and the pulmonary arteries (PA) are depicted in blue. The dorsal mid-sagittal SHF population has moved to the left and has extended a large band of cells to the left (pulmonary) side (P) of the OFT extending along the pulmonary vessel wall and the myocardial OFT (A,B,E, black arrowhead, F). The flow divider (FD) has separated the 6th pharyngeal arch arteries and has also moved to the left (E,F). The Tfap2α-positive NCCs (G) are encircled with a striped line. They have taken up a right-sided position next to the FD and are connected to the NCCs in the opposing endocardial cushions (ec). The NCC is forming the aorto-pulmonary septum (E-G, double-headed arrow), viewed in the 3D reconstruction (B) at the connection between dark and bright green. In the knockout embryo, both the SHF and the NCC population on the right (aortic) side are not well developed (C). Because the dorsal SHF population (not shown in the reconstruction) remains in midline (Nkx2.5-positive cells in sections H,I), there is hardly an extension along the left (pulmonary) side (C,D and H,I, black arrowhead). There is a marked Tfap2α (J) NCC population, however, on the left (pulmonary) side that connects to the bright green NCCs in the endocardial cushions. This does not lead to septation of the aortic sac, which now has become a common arterial trunk (CAT) (H-J). The FD is still prominently present and is Nkx2.5 (SHF)-positive (H,I, arrow) and Tfap2α (NCC)-negative (J; arrow). Section levels have been indicated in the reconstructions (A,C). Scale bars: 50 µm. See also the online supplemental videos (E11.5_video_1080p.mpg) and interactive pdf reconstructions (E11.5_WT_revise.pdf and E11.5_KO_revise.pdf) on http://www.caskanatomy.info/research/supplement_megalin.html . S, superior; R, right; L, left; I, inferior; A, anterior; P, posterior; Pt, pulmonary trunk.

Article Snippet: After deparaffination and rehydration of the slides, immunohistochemical staining was performed with the antibodies anti-atrial myosin light chain 2a (MLC-2a) as a myocardial marker (1/4000, kindly provided by S. W. Kubalak, Medical University of South Carolina, SC), anti-Nkx2.5 as a (pre)myocardial marker and a marker for SHF-derived cells (1/4000, Santa Cruz Biotechnology, Inc., CA, SC-8697), anti-LPR2 (1/800, kindly provided by J. Herz, University of Texas Southwestern Medical Center, TX), anti-WT1 as a marker for the epicardium and early migrating EPDCs (1/500, Calbiochem, ONCOCA1026), anti-Tfap2α as an NCC marker (1/500, GeneTex Inc., GTX62588) and anti-CD31 (M-20) as a marker for endothelium and endocardium (1/4000, Santa Cruz Biotechnology, Inc., CA, SC-1506).

Techniques: Knock-Out