The Experts below are selected from a list of 477 Experts worldwide ranked by ideXlab platform
Sarah J Childs - One of the best experts on this subject based on the ideXlab platform.
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an α smooth muscle actin acta2 αsma zebrafish transgenic line marking vascular mural cells and visceral smooth muscle cells
PLOS ONE, 2014Co-Authors: Thomas Whitesell, Massimo Santoro, Regan M Kennedy, Alyson D Carter, Evvilynn Rollins, Sonja Georgijevic, Sarah J ChildsAbstract:Mural cells of the vascular system include vascular smooth muscle cells (SMCs) and pericytes whose role is to stabilize and/or provide contractility to blood vessels. One of the earliest markers of mural cell development in vertebrates is α smooth muscle actin (acta2; αsma), which is expressed by pericytes and SMCs. In vivo models of vascular mural cell development in zebrafish are currently lacking, therefore we developed two transgenic zebrafish lines driving expression of GFP or mCherry in acta2-expressing cells. These transgenic fish were used to trace the live development of mural cells in embryonic and larval transgenic zebrafish. acta2:EGFP transgenic animals show expression that largely mirrors native acta2 expression, with early pan-muscle expression starting at 24 hpf in the heart muscle, followed by skeletal and visceral muscle. At 3.5 dpf, expression in the Bulbus arteriosus and ventral aorta marks the first expression in vascular smooth muscle. Over the next 10 days of development, the number of acta2:EGFP positive cells and the number of types of blood vessels associated with mural cells increases. Interestingly, the mural cells are not motile and remain in the same position once they express the acta2:EGFP transgene. Taken together, our data suggests that zebrafish mural cells develop relatively late, and have little mobility once they associate with vessels.
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An α-smooth muscle actin (acta2/αsma) zebrafish transgenic line marking vascular mural cells and visceral smooth muscle cells.
PloS one, 2014Co-Authors: Thomas Whitesell, Massimo Santoro, Regan M Kennedy, Alyson D Carter, Evvilynn Rollins, Sonja Georgijevic, Sarah J ChildsAbstract:Mural cells of the vascular system include vascular smooth muscle cells (SMCs) and pericytes whose role is to stabilize and/or provide contractility to blood vessels. One of the earliest markers of mural cell development in vertebrates is α smooth muscle actin (acta2; αsma), which is expressed by pericytes and SMCs. In vivo models of vascular mural cell development in zebrafish are currently lacking, therefore we developed two transgenic zebrafish lines driving expression of GFP or mCherry in acta2-expressing cells. These transgenic fish were used to trace the live development of mural cells in embryonic and larval transgenic zebrafish. acta2:EGFP transgenic animals show expression that largely mirrors native acta2 expression, with early pan-muscle expression starting at 24 hpf in the heart muscle, followed by skeletal and visceral muscle. At 3.5 dpf, expression in the Bulbus arteriosus and ventral aorta marks the first expression in vascular smooth muscle. Over the next 10 days of development, the number of acta2:EGFP positive cells and the number of types of blood vessels associated with mural cells increases. Interestingly, the mural cells are not motile and remain in the same position once they express the acta2:EGFP transgene. Taken together, our data suggests that zebrafish mural cells develop relatively late, and have little mobility once they associate with vessels.
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Mural cell and endothelial development in the ventral head of larval zebrafish.
2014Co-Authors: Thomas R. Whitesell, Regan M Kennedy, Alyson D Carter, Evvilynn Rollins, Sonja Georgijevic, Massimo M. Santoro, Sarah J ChildsAbstract:Confocal micrographs collected from a ventral point of view show a progressive increase in vessel complexity (red, A, C, E, G) and in density of mural cell coverage of aortic arch vessels (green, B, D, F, H) from 4 dpf (A, B), 7 dpf (C, D), 11 dpf (E, F) through 14 dpf (G, H). Heart expression of acta2:EGFP is maintained. aaa = aortic arch arteries; va = ventral aorta; ba = Bulbus arteriosus. Scale bar in A represents 100 µm.
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Smooth muscle markers are restricted to the developing cardiac outflow tract by 56 hpf.
2014Co-Authors: Thomas R. Whitesell, Regan M Kennedy, Alyson D Carter, Evvilynn Rollins, Sonja Georgijevic, Massimo M. Santoro, Sarah J ChildsAbstract:(A) At 56 hpf, acta2 expression is restricted to the developing BA. (B,C) Double transgenic Tg(acta2:EGFP)ca7; Tg(kdrl:mCherry)ci5 embryo shows expression of EGFP in both the atrium and ventricle of the heart at 56 hpf, but not in the BA. (D) acta2 expression is evident at 78 hpf in the BA in both wholemount and cross section (E) and in transgenic animals (F). (G–I) Expression of acta2 continues to be restricted to the BA and ventral aorta (VA) at 100 hpf by in situ hybridization and in transgenic fish. (J–O): Cross sections of the 22 dpf BA show a multilamellar arterial phenotype as visualized by hematoxylin and eosin staining (J), in situ hybridization of acta2 (K) and transgenic GFP (nuclei stained blue with DAPI, L). The Bulbus vascular wall consists of three layers: an inner intima, middle media, and outer adventitia (Ad, separated by black lines in J). The intima is endothelial (arrowheads point to nuclei of endothelial cells). The media consists of 3–4 cell-thick layers of vascular smooth muscle cells (M, arrows point to nuclei of SMCs). In comparison to the BA, the vascular wall of the VA at 22 dpf is thin (M) but expresses acta2 by in situ hybridization (N) and GFP in transgenic animals (O). The endothelium of VA is covered by a thin layer of SMCs (arrowheads point to nuclei of SMCs). (P) In situ hybridization of the wholemount adult heart shows strong staining in the Bulbus arteriosus, but not ventricle or atrium, which is localized to the myocardial wall in cross section (Q). (R) Wholemount dissected acta2:EGFP transgenic heart shows stronger expression of GFP in the Bulbus arteriosus as compared to ventricle. Staining is also continuous with the ventral aorta. In B,C, F, I, and R, green expression is acta2:EGFP transgene. Scale bar in B, C, F, and I is 100 µm. Scale bar in E, H, and Q is 50 µm. Scale bar in K, L, N, and O is 20 µm.
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Acta2 promoter/enhancer construct design and expression in zebrafish.
2014Co-Authors: Thomas R. Whitesell, Regan M Kennedy, Alyson D Carter, Evvilynn Rollins, Sonja Georgijevic, Massimo M. Santoro, Sarah J ChildsAbstract:(A) A zebrafish (Dr) enhancer/promoter construct was constructed from the proximal promoter and first intron sequence of the zebrafish acta2 gene, and contains three highly conserved CArG binding sites also found in the mouse (Mm) acta2 proximal promoter and first intron. (B) Comparison of zebrafish CaRG boxes A and B in zebrafish, tilapia and medaka. (C,D) By wholemount in situ hybridization, acta2 shows strong expression in the gut (g) at 72 hpf (B), and expressed in the gut, swim bladder (sb), ventral aorta (va), floor plate (fp), aortic arch arteries (aaa), and Bulbus arteriosus (ba) at 100 hpf (C). (E,F) Co-localization of wholemount in situ hybridization acta2 and anti-GFP staining of the acta2:GFP transgene shows strong expression in the aortic arch arteries (aaa) at 100 hpf. (G,H,I) 4 dpf acta2:EGFP transgenic fish (H) stained with Tagln rabbit polyclonal antibody (G). Merge (I) shows co-localization between acta2:GFP and Tagln. Arrowheads in G–I depict vascular mural cells. Scale bar in G represents 20 µm.
Thomas Whitesell - One of the best experts on this subject based on the ideXlab platform.
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an α smooth muscle actin acta2 αsma zebrafish transgenic line marking vascular mural cells and visceral smooth muscle cells
PLOS ONE, 2014Co-Authors: Thomas Whitesell, Massimo Santoro, Regan M Kennedy, Alyson D Carter, Evvilynn Rollins, Sonja Georgijevic, Sarah J ChildsAbstract:Mural cells of the vascular system include vascular smooth muscle cells (SMCs) and pericytes whose role is to stabilize and/or provide contractility to blood vessels. One of the earliest markers of mural cell development in vertebrates is α smooth muscle actin (acta2; αsma), which is expressed by pericytes and SMCs. In vivo models of vascular mural cell development in zebrafish are currently lacking, therefore we developed two transgenic zebrafish lines driving expression of GFP or mCherry in acta2-expressing cells. These transgenic fish were used to trace the live development of mural cells in embryonic and larval transgenic zebrafish. acta2:EGFP transgenic animals show expression that largely mirrors native acta2 expression, with early pan-muscle expression starting at 24 hpf in the heart muscle, followed by skeletal and visceral muscle. At 3.5 dpf, expression in the Bulbus arteriosus and ventral aorta marks the first expression in vascular smooth muscle. Over the next 10 days of development, the number of acta2:EGFP positive cells and the number of types of blood vessels associated with mural cells increases. Interestingly, the mural cells are not motile and remain in the same position once they express the acta2:EGFP transgene. Taken together, our data suggests that zebrafish mural cells develop relatively late, and have little mobility once they associate with vessels.
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An α-smooth muscle actin (acta2/αsma) zebrafish transgenic line marking vascular mural cells and visceral smooth muscle cells.
PloS one, 2014Co-Authors: Thomas Whitesell, Massimo Santoro, Regan M Kennedy, Alyson D Carter, Evvilynn Rollins, Sonja Georgijevic, Sarah J ChildsAbstract:Mural cells of the vascular system include vascular smooth muscle cells (SMCs) and pericytes whose role is to stabilize and/or provide contractility to blood vessels. One of the earliest markers of mural cell development in vertebrates is α smooth muscle actin (acta2; αsma), which is expressed by pericytes and SMCs. In vivo models of vascular mural cell development in zebrafish are currently lacking, therefore we developed two transgenic zebrafish lines driving expression of GFP or mCherry in acta2-expressing cells. These transgenic fish were used to trace the live development of mural cells in embryonic and larval transgenic zebrafish. acta2:EGFP transgenic animals show expression that largely mirrors native acta2 expression, with early pan-muscle expression starting at 24 hpf in the heart muscle, followed by skeletal and visceral muscle. At 3.5 dpf, expression in the Bulbus arteriosus and ventral aorta marks the first expression in vascular smooth muscle. Over the next 10 days of development, the number of acta2:EGFP positive cells and the number of types of blood vessels associated with mural cells increases. Interestingly, the mural cells are not motile and remain in the same position once they express the acta2:EGFP transgene. Taken together, our data suggests that zebrafish mural cells develop relatively late, and have little mobility once they associate with vessels.
Regan M Kennedy - One of the best experts on this subject based on the ideXlab platform.
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an α smooth muscle actin acta2 αsma zebrafish transgenic line marking vascular mural cells and visceral smooth muscle cells
PLOS ONE, 2014Co-Authors: Thomas Whitesell, Massimo Santoro, Regan M Kennedy, Alyson D Carter, Evvilynn Rollins, Sonja Georgijevic, Sarah J ChildsAbstract:Mural cells of the vascular system include vascular smooth muscle cells (SMCs) and pericytes whose role is to stabilize and/or provide contractility to blood vessels. One of the earliest markers of mural cell development in vertebrates is α smooth muscle actin (acta2; αsma), which is expressed by pericytes and SMCs. In vivo models of vascular mural cell development in zebrafish are currently lacking, therefore we developed two transgenic zebrafish lines driving expression of GFP or mCherry in acta2-expressing cells. These transgenic fish were used to trace the live development of mural cells in embryonic and larval transgenic zebrafish. acta2:EGFP transgenic animals show expression that largely mirrors native acta2 expression, with early pan-muscle expression starting at 24 hpf in the heart muscle, followed by skeletal and visceral muscle. At 3.5 dpf, expression in the Bulbus arteriosus and ventral aorta marks the first expression in vascular smooth muscle. Over the next 10 days of development, the number of acta2:EGFP positive cells and the number of types of blood vessels associated with mural cells increases. Interestingly, the mural cells are not motile and remain in the same position once they express the acta2:EGFP transgene. Taken together, our data suggests that zebrafish mural cells develop relatively late, and have little mobility once they associate with vessels.
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An α-smooth muscle actin (acta2/αsma) zebrafish transgenic line marking vascular mural cells and visceral smooth muscle cells.
PloS one, 2014Co-Authors: Thomas Whitesell, Massimo Santoro, Regan M Kennedy, Alyson D Carter, Evvilynn Rollins, Sonja Georgijevic, Sarah J ChildsAbstract:Mural cells of the vascular system include vascular smooth muscle cells (SMCs) and pericytes whose role is to stabilize and/or provide contractility to blood vessels. One of the earliest markers of mural cell development in vertebrates is α smooth muscle actin (acta2; αsma), which is expressed by pericytes and SMCs. In vivo models of vascular mural cell development in zebrafish are currently lacking, therefore we developed two transgenic zebrafish lines driving expression of GFP or mCherry in acta2-expressing cells. These transgenic fish were used to trace the live development of mural cells in embryonic and larval transgenic zebrafish. acta2:EGFP transgenic animals show expression that largely mirrors native acta2 expression, with early pan-muscle expression starting at 24 hpf in the heart muscle, followed by skeletal and visceral muscle. At 3.5 dpf, expression in the Bulbus arteriosus and ventral aorta marks the first expression in vascular smooth muscle. Over the next 10 days of development, the number of acta2:EGFP positive cells and the number of types of blood vessels associated with mural cells increases. Interestingly, the mural cells are not motile and remain in the same position once they express the acta2:EGFP transgene. Taken together, our data suggests that zebrafish mural cells develop relatively late, and have little mobility once they associate with vessels.
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Mural cell and endothelial development in the ventral head of larval zebrafish.
2014Co-Authors: Thomas R. Whitesell, Regan M Kennedy, Alyson D Carter, Evvilynn Rollins, Sonja Georgijevic, Massimo M. Santoro, Sarah J ChildsAbstract:Confocal micrographs collected from a ventral point of view show a progressive increase in vessel complexity (red, A, C, E, G) and in density of mural cell coverage of aortic arch vessels (green, B, D, F, H) from 4 dpf (A, B), 7 dpf (C, D), 11 dpf (E, F) through 14 dpf (G, H). Heart expression of acta2:EGFP is maintained. aaa = aortic arch arteries; va = ventral aorta; ba = Bulbus arteriosus. Scale bar in A represents 100 µm.
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Smooth muscle markers are restricted to the developing cardiac outflow tract by 56 hpf.
2014Co-Authors: Thomas R. Whitesell, Regan M Kennedy, Alyson D Carter, Evvilynn Rollins, Sonja Georgijevic, Massimo M. Santoro, Sarah J ChildsAbstract:(A) At 56 hpf, acta2 expression is restricted to the developing BA. (B,C) Double transgenic Tg(acta2:EGFP)ca7; Tg(kdrl:mCherry)ci5 embryo shows expression of EGFP in both the atrium and ventricle of the heart at 56 hpf, but not in the BA. (D) acta2 expression is evident at 78 hpf in the BA in both wholemount and cross section (E) and in transgenic animals (F). (G–I) Expression of acta2 continues to be restricted to the BA and ventral aorta (VA) at 100 hpf by in situ hybridization and in transgenic fish. (J–O): Cross sections of the 22 dpf BA show a multilamellar arterial phenotype as visualized by hematoxylin and eosin staining (J), in situ hybridization of acta2 (K) and transgenic GFP (nuclei stained blue with DAPI, L). The Bulbus vascular wall consists of three layers: an inner intima, middle media, and outer adventitia (Ad, separated by black lines in J). The intima is endothelial (arrowheads point to nuclei of endothelial cells). The media consists of 3–4 cell-thick layers of vascular smooth muscle cells (M, arrows point to nuclei of SMCs). In comparison to the BA, the vascular wall of the VA at 22 dpf is thin (M) but expresses acta2 by in situ hybridization (N) and GFP in transgenic animals (O). The endothelium of VA is covered by a thin layer of SMCs (arrowheads point to nuclei of SMCs). (P) In situ hybridization of the wholemount adult heart shows strong staining in the Bulbus arteriosus, but not ventricle or atrium, which is localized to the myocardial wall in cross section (Q). (R) Wholemount dissected acta2:EGFP transgenic heart shows stronger expression of GFP in the Bulbus arteriosus as compared to ventricle. Staining is also continuous with the ventral aorta. In B,C, F, I, and R, green expression is acta2:EGFP transgene. Scale bar in B, C, F, and I is 100 µm. Scale bar in E, H, and Q is 50 µm. Scale bar in K, L, N, and O is 20 µm.
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Acta2 promoter/enhancer construct design and expression in zebrafish.
2014Co-Authors: Thomas R. Whitesell, Regan M Kennedy, Alyson D Carter, Evvilynn Rollins, Sonja Georgijevic, Massimo M. Santoro, Sarah J ChildsAbstract:(A) A zebrafish (Dr) enhancer/promoter construct was constructed from the proximal promoter and first intron sequence of the zebrafish acta2 gene, and contains three highly conserved CArG binding sites also found in the mouse (Mm) acta2 proximal promoter and first intron. (B) Comparison of zebrafish CaRG boxes A and B in zebrafish, tilapia and medaka. (C,D) By wholemount in situ hybridization, acta2 shows strong expression in the gut (g) at 72 hpf (B), and expressed in the gut, swim bladder (sb), ventral aorta (va), floor plate (fp), aortic arch arteries (aaa), and Bulbus arteriosus (ba) at 100 hpf (C). (E,F) Co-localization of wholemount in situ hybridization acta2 and anti-GFP staining of the acta2:GFP transgene shows strong expression in the aortic arch arteries (aaa) at 100 hpf. (G,H,I) 4 dpf acta2:EGFP transgenic fish (H) stained with Tagln rabbit polyclonal antibody (G). Merge (I) shows co-localization between acta2:GFP and Tagln. Arrowheads in G–I depict vascular mural cells. Scale bar in G represents 20 µm.
Alyson D Carter - One of the best experts on this subject based on the ideXlab platform.
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an α smooth muscle actin acta2 αsma zebrafish transgenic line marking vascular mural cells and visceral smooth muscle cells
PLOS ONE, 2014Co-Authors: Thomas Whitesell, Massimo Santoro, Regan M Kennedy, Alyson D Carter, Evvilynn Rollins, Sonja Georgijevic, Sarah J ChildsAbstract:Mural cells of the vascular system include vascular smooth muscle cells (SMCs) and pericytes whose role is to stabilize and/or provide contractility to blood vessels. One of the earliest markers of mural cell development in vertebrates is α smooth muscle actin (acta2; αsma), which is expressed by pericytes and SMCs. In vivo models of vascular mural cell development in zebrafish are currently lacking, therefore we developed two transgenic zebrafish lines driving expression of GFP or mCherry in acta2-expressing cells. These transgenic fish were used to trace the live development of mural cells in embryonic and larval transgenic zebrafish. acta2:EGFP transgenic animals show expression that largely mirrors native acta2 expression, with early pan-muscle expression starting at 24 hpf in the heart muscle, followed by skeletal and visceral muscle. At 3.5 dpf, expression in the Bulbus arteriosus and ventral aorta marks the first expression in vascular smooth muscle. Over the next 10 days of development, the number of acta2:EGFP positive cells and the number of types of blood vessels associated with mural cells increases. Interestingly, the mural cells are not motile and remain in the same position once they express the acta2:EGFP transgene. Taken together, our data suggests that zebrafish mural cells develop relatively late, and have little mobility once they associate with vessels.
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An α-smooth muscle actin (acta2/αsma) zebrafish transgenic line marking vascular mural cells and visceral smooth muscle cells.
PloS one, 2014Co-Authors: Thomas Whitesell, Massimo Santoro, Regan M Kennedy, Alyson D Carter, Evvilynn Rollins, Sonja Georgijevic, Sarah J ChildsAbstract:Mural cells of the vascular system include vascular smooth muscle cells (SMCs) and pericytes whose role is to stabilize and/or provide contractility to blood vessels. One of the earliest markers of mural cell development in vertebrates is α smooth muscle actin (acta2; αsma), which is expressed by pericytes and SMCs. In vivo models of vascular mural cell development in zebrafish are currently lacking, therefore we developed two transgenic zebrafish lines driving expression of GFP or mCherry in acta2-expressing cells. These transgenic fish were used to trace the live development of mural cells in embryonic and larval transgenic zebrafish. acta2:EGFP transgenic animals show expression that largely mirrors native acta2 expression, with early pan-muscle expression starting at 24 hpf in the heart muscle, followed by skeletal and visceral muscle. At 3.5 dpf, expression in the Bulbus arteriosus and ventral aorta marks the first expression in vascular smooth muscle. Over the next 10 days of development, the number of acta2:EGFP positive cells and the number of types of blood vessels associated with mural cells increases. Interestingly, the mural cells are not motile and remain in the same position once they express the acta2:EGFP transgene. Taken together, our data suggests that zebrafish mural cells develop relatively late, and have little mobility once they associate with vessels.
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Mural cell and endothelial development in the ventral head of larval zebrafish.
2014Co-Authors: Thomas R. Whitesell, Regan M Kennedy, Alyson D Carter, Evvilynn Rollins, Sonja Georgijevic, Massimo M. Santoro, Sarah J ChildsAbstract:Confocal micrographs collected from a ventral point of view show a progressive increase in vessel complexity (red, A, C, E, G) and in density of mural cell coverage of aortic arch vessels (green, B, D, F, H) from 4 dpf (A, B), 7 dpf (C, D), 11 dpf (E, F) through 14 dpf (G, H). Heart expression of acta2:EGFP is maintained. aaa = aortic arch arteries; va = ventral aorta; ba = Bulbus arteriosus. Scale bar in A represents 100 µm.
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Smooth muscle markers are restricted to the developing cardiac outflow tract by 56 hpf.
2014Co-Authors: Thomas R. Whitesell, Regan M Kennedy, Alyson D Carter, Evvilynn Rollins, Sonja Georgijevic, Massimo M. Santoro, Sarah J ChildsAbstract:(A) At 56 hpf, acta2 expression is restricted to the developing BA. (B,C) Double transgenic Tg(acta2:EGFP)ca7; Tg(kdrl:mCherry)ci5 embryo shows expression of EGFP in both the atrium and ventricle of the heart at 56 hpf, but not in the BA. (D) acta2 expression is evident at 78 hpf in the BA in both wholemount and cross section (E) and in transgenic animals (F). (G–I) Expression of acta2 continues to be restricted to the BA and ventral aorta (VA) at 100 hpf by in situ hybridization and in transgenic fish. (J–O): Cross sections of the 22 dpf BA show a multilamellar arterial phenotype as visualized by hematoxylin and eosin staining (J), in situ hybridization of acta2 (K) and transgenic GFP (nuclei stained blue with DAPI, L). The Bulbus vascular wall consists of three layers: an inner intima, middle media, and outer adventitia (Ad, separated by black lines in J). The intima is endothelial (arrowheads point to nuclei of endothelial cells). The media consists of 3–4 cell-thick layers of vascular smooth muscle cells (M, arrows point to nuclei of SMCs). In comparison to the BA, the vascular wall of the VA at 22 dpf is thin (M) but expresses acta2 by in situ hybridization (N) and GFP in transgenic animals (O). The endothelium of VA is covered by a thin layer of SMCs (arrowheads point to nuclei of SMCs). (P) In situ hybridization of the wholemount adult heart shows strong staining in the Bulbus arteriosus, but not ventricle or atrium, which is localized to the myocardial wall in cross section (Q). (R) Wholemount dissected acta2:EGFP transgenic heart shows stronger expression of GFP in the Bulbus arteriosus as compared to ventricle. Staining is also continuous with the ventral aorta. In B,C, F, I, and R, green expression is acta2:EGFP transgene. Scale bar in B, C, F, and I is 100 µm. Scale bar in E, H, and Q is 50 µm. Scale bar in K, L, N, and O is 20 µm.
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Acta2 promoter/enhancer construct design and expression in zebrafish.
2014Co-Authors: Thomas R. Whitesell, Regan M Kennedy, Alyson D Carter, Evvilynn Rollins, Sonja Georgijevic, Massimo M. Santoro, Sarah J ChildsAbstract:(A) A zebrafish (Dr) enhancer/promoter construct was constructed from the proximal promoter and first intron sequence of the zebrafish acta2 gene, and contains three highly conserved CArG binding sites also found in the mouse (Mm) acta2 proximal promoter and first intron. (B) Comparison of zebrafish CaRG boxes A and B in zebrafish, tilapia and medaka. (C,D) By wholemount in situ hybridization, acta2 shows strong expression in the gut (g) at 72 hpf (B), and expressed in the gut, swim bladder (sb), ventral aorta (va), floor plate (fp), aortic arch arteries (aaa), and Bulbus arteriosus (ba) at 100 hpf (C). (E,F) Co-localization of wholemount in situ hybridization acta2 and anti-GFP staining of the acta2:GFP transgene shows strong expression in the aortic arch arteries (aaa) at 100 hpf. (G,H,I) 4 dpf acta2:EGFP transgenic fish (H) stained with Tagln rabbit polyclonal antibody (G). Merge (I) shows co-localization between acta2:GFP and Tagln. Arrowheads in G–I depict vascular mural cells. Scale bar in G represents 20 µm.
Sonja Georgijevic - One of the best experts on this subject based on the ideXlab platform.
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an α smooth muscle actin acta2 αsma zebrafish transgenic line marking vascular mural cells and visceral smooth muscle cells
PLOS ONE, 2014Co-Authors: Thomas Whitesell, Massimo Santoro, Regan M Kennedy, Alyson D Carter, Evvilynn Rollins, Sonja Georgijevic, Sarah J ChildsAbstract:Mural cells of the vascular system include vascular smooth muscle cells (SMCs) and pericytes whose role is to stabilize and/or provide contractility to blood vessels. One of the earliest markers of mural cell development in vertebrates is α smooth muscle actin (acta2; αsma), which is expressed by pericytes and SMCs. In vivo models of vascular mural cell development in zebrafish are currently lacking, therefore we developed two transgenic zebrafish lines driving expression of GFP or mCherry in acta2-expressing cells. These transgenic fish were used to trace the live development of mural cells in embryonic and larval transgenic zebrafish. acta2:EGFP transgenic animals show expression that largely mirrors native acta2 expression, with early pan-muscle expression starting at 24 hpf in the heart muscle, followed by skeletal and visceral muscle. At 3.5 dpf, expression in the Bulbus arteriosus and ventral aorta marks the first expression in vascular smooth muscle. Over the next 10 days of development, the number of acta2:EGFP positive cells and the number of types of blood vessels associated with mural cells increases. Interestingly, the mural cells are not motile and remain in the same position once they express the acta2:EGFP transgene. Taken together, our data suggests that zebrafish mural cells develop relatively late, and have little mobility once they associate with vessels.
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An α-smooth muscle actin (acta2/αsma) zebrafish transgenic line marking vascular mural cells and visceral smooth muscle cells.
PloS one, 2014Co-Authors: Thomas Whitesell, Massimo Santoro, Regan M Kennedy, Alyson D Carter, Evvilynn Rollins, Sonja Georgijevic, Sarah J ChildsAbstract:Mural cells of the vascular system include vascular smooth muscle cells (SMCs) and pericytes whose role is to stabilize and/or provide contractility to blood vessels. One of the earliest markers of mural cell development in vertebrates is α smooth muscle actin (acta2; αsma), which is expressed by pericytes and SMCs. In vivo models of vascular mural cell development in zebrafish are currently lacking, therefore we developed two transgenic zebrafish lines driving expression of GFP or mCherry in acta2-expressing cells. These transgenic fish were used to trace the live development of mural cells in embryonic and larval transgenic zebrafish. acta2:EGFP transgenic animals show expression that largely mirrors native acta2 expression, with early pan-muscle expression starting at 24 hpf in the heart muscle, followed by skeletal and visceral muscle. At 3.5 dpf, expression in the Bulbus arteriosus and ventral aorta marks the first expression in vascular smooth muscle. Over the next 10 days of development, the number of acta2:EGFP positive cells and the number of types of blood vessels associated with mural cells increases. Interestingly, the mural cells are not motile and remain in the same position once they express the acta2:EGFP transgene. Taken together, our data suggests that zebrafish mural cells develop relatively late, and have little mobility once they associate with vessels.
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Mural cell and endothelial development in the ventral head of larval zebrafish.
2014Co-Authors: Thomas R. Whitesell, Regan M Kennedy, Alyson D Carter, Evvilynn Rollins, Sonja Georgijevic, Massimo M. Santoro, Sarah J ChildsAbstract:Confocal micrographs collected from a ventral point of view show a progressive increase in vessel complexity (red, A, C, E, G) and in density of mural cell coverage of aortic arch vessels (green, B, D, F, H) from 4 dpf (A, B), 7 dpf (C, D), 11 dpf (E, F) through 14 dpf (G, H). Heart expression of acta2:EGFP is maintained. aaa = aortic arch arteries; va = ventral aorta; ba = Bulbus arteriosus. Scale bar in A represents 100 µm.
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Smooth muscle markers are restricted to the developing cardiac outflow tract by 56 hpf.
2014Co-Authors: Thomas R. Whitesell, Regan M Kennedy, Alyson D Carter, Evvilynn Rollins, Sonja Georgijevic, Massimo M. Santoro, Sarah J ChildsAbstract:(A) At 56 hpf, acta2 expression is restricted to the developing BA. (B,C) Double transgenic Tg(acta2:EGFP)ca7; Tg(kdrl:mCherry)ci5 embryo shows expression of EGFP in both the atrium and ventricle of the heart at 56 hpf, but not in the BA. (D) acta2 expression is evident at 78 hpf in the BA in both wholemount and cross section (E) and in transgenic animals (F). (G–I) Expression of acta2 continues to be restricted to the BA and ventral aorta (VA) at 100 hpf by in situ hybridization and in transgenic fish. (J–O): Cross sections of the 22 dpf BA show a multilamellar arterial phenotype as visualized by hematoxylin and eosin staining (J), in situ hybridization of acta2 (K) and transgenic GFP (nuclei stained blue with DAPI, L). The Bulbus vascular wall consists of three layers: an inner intima, middle media, and outer adventitia (Ad, separated by black lines in J). The intima is endothelial (arrowheads point to nuclei of endothelial cells). The media consists of 3–4 cell-thick layers of vascular smooth muscle cells (M, arrows point to nuclei of SMCs). In comparison to the BA, the vascular wall of the VA at 22 dpf is thin (M) but expresses acta2 by in situ hybridization (N) and GFP in transgenic animals (O). The endothelium of VA is covered by a thin layer of SMCs (arrowheads point to nuclei of SMCs). (P) In situ hybridization of the wholemount adult heart shows strong staining in the Bulbus arteriosus, but not ventricle or atrium, which is localized to the myocardial wall in cross section (Q). (R) Wholemount dissected acta2:EGFP transgenic heart shows stronger expression of GFP in the Bulbus arteriosus as compared to ventricle. Staining is also continuous with the ventral aorta. In B,C, F, I, and R, green expression is acta2:EGFP transgene. Scale bar in B, C, F, and I is 100 µm. Scale bar in E, H, and Q is 50 µm. Scale bar in K, L, N, and O is 20 µm.
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Acta2 promoter/enhancer construct design and expression in zebrafish.
2014Co-Authors: Thomas R. Whitesell, Regan M Kennedy, Alyson D Carter, Evvilynn Rollins, Sonja Georgijevic, Massimo M. Santoro, Sarah J ChildsAbstract:(A) A zebrafish (Dr) enhancer/promoter construct was constructed from the proximal promoter and first intron sequence of the zebrafish acta2 gene, and contains three highly conserved CArG binding sites also found in the mouse (Mm) acta2 proximal promoter and first intron. (B) Comparison of zebrafish CaRG boxes A and B in zebrafish, tilapia and medaka. (C,D) By wholemount in situ hybridization, acta2 shows strong expression in the gut (g) at 72 hpf (B), and expressed in the gut, swim bladder (sb), ventral aorta (va), floor plate (fp), aortic arch arteries (aaa), and Bulbus arteriosus (ba) at 100 hpf (C). (E,F) Co-localization of wholemount in situ hybridization acta2 and anti-GFP staining of the acta2:GFP transgene shows strong expression in the aortic arch arteries (aaa) at 100 hpf. (G,H,I) 4 dpf acta2:EGFP transgenic fish (H) stained with Tagln rabbit polyclonal antibody (G). Merge (I) shows co-localization between acta2:GFP and Tagln. Arrowheads in G–I depict vascular mural cells. Scale bar in G represents 20 µm.
