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Conditional expression of Spry1 in neural crest causes craniofacial and cardiac defects.

Yang X, Kilgallen S, Andreeva V, Spicer DB, Pinz I, Friesel R - BMC Dev. Biol. (2010)

Bottom Line: Spry1;Wnt1-Cre embryos die perinatally and exhibit facial clefting, cleft palate, cardiac and cranial nerve defects.These defects appear to be the result of decreased proliferation and increased apoptosis of neural crest and neural crest-derived cell populations.In addition, the domains of expression of several key transcription factors important to normal craniofacial and cardiac development including AP2, Msx2, Dlx5, and Dlx6 were reduced in Spry1;Wnt1-Cre transgenic embryos.

View Article: PubMed Central - HTML - PubMed

Affiliation: Center for Molecular Medicine, Maine Medical Center Research Institute, Scarborough, ME 04074, USA.

ABSTRACT

Background: Growth factors and their receptors are mediators of organogenesis and must be tightly regulated in a temporal and spatial manner for proper tissue morphogenesis. Intracellular regulators of growth factor signaling pathways provide an additional level of control. Members of the Sprouty family negatively regulate receptor tyrosine kinase pathways in several developmental contexts. To gain insight into the role of Spry1 in neural crest development, we analyzed the developmental effects of conditional expression of Spry1 in neural crest-derived tissues.

Results: Here we report that conditional expression of Spry1 in neural crest cells causes defects in craniofacial and cardiac development in mice. Spry1;Wnt1-Cre embryos die perinatally and exhibit facial clefting, cleft palate, cardiac and cranial nerve defects. These defects appear to be the result of decreased proliferation and increased apoptosis of neural crest and neural crest-derived cell populations. In addition, the domains of expression of several key transcription factors important to normal craniofacial and cardiac development including AP2, Msx2, Dlx5, and Dlx6 were reduced in Spry1;Wnt1-Cre transgenic embryos.

Conclusion: Collectively, these data suggest that Spry1 is an important regulator of craniofacial and cardiac morphogenesis and perturbations in Spry1 levels may contribute to congenital disorders involving tissues of neural crest origin.

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Spry1;Wnt1-Cre embryos show outflow tract defects at E9.5. (A, B) Whole mount β-gal staining of Cre-negative littermate control and Spry1;Wnt1-Cre E9.5 embryos. (A) WT shows intense β-gal staining of the outflow tract (white arrow) and branchial arches, indicating cells of NC origin. (B) Spry1;Wnt1-Cre E9.5 embryo, shows variable β-gal staining of the pharyngeal arches and reduced staining of the outflow tract (white arrowhead). (C, D) Sagittal sections of whole mount embryos with nuclear fast red counter-staining. (C) WT shows normal distribution of cardiac NCC within the outflow tract, with β-galactosidase positive NCC cells extending down to the bulbis cordis. Panel D, Spry1;Wnt1-Cre reveals outflow tract with reduced β-galactosidase positive NCC cells. In addition the first branchial arch, mandibular component, is greatly reduced in size relative to the WT. The outflow tract in Spry1;Wnt1-Cre embryos is shortened and does not adopt the spiral configuration as seen in the WT. (E,F) High power images of C,D; white boxed areas indicate field of view. (E) WT, black arrow indicates cardiac NCC contributing cardiac mesenchyme. (F) Spry1;Wnt1-Cre embryo, black arrowhead notes paucity of NCC in cardiac mesenchyme. FBA: first branchial arch, mandibular component. Data are representative of six embryos from each group.
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Figure 10: Spry1;Wnt1-Cre embryos show outflow tract defects at E9.5. (A, B) Whole mount β-gal staining of Cre-negative littermate control and Spry1;Wnt1-Cre E9.5 embryos. (A) WT shows intense β-gal staining of the outflow tract (white arrow) and branchial arches, indicating cells of NC origin. (B) Spry1;Wnt1-Cre E9.5 embryo, shows variable β-gal staining of the pharyngeal arches and reduced staining of the outflow tract (white arrowhead). (C, D) Sagittal sections of whole mount embryos with nuclear fast red counter-staining. (C) WT shows normal distribution of cardiac NCC within the outflow tract, with β-galactosidase positive NCC cells extending down to the bulbis cordis. Panel D, Spry1;Wnt1-Cre reveals outflow tract with reduced β-galactosidase positive NCC cells. In addition the first branchial arch, mandibular component, is greatly reduced in size relative to the WT. The outflow tract in Spry1;Wnt1-Cre embryos is shortened and does not adopt the spiral configuration as seen in the WT. (E,F) High power images of C,D; white boxed areas indicate field of view. (E) WT, black arrow indicates cardiac NCC contributing cardiac mesenchyme. (F) Spry1;Wnt1-Cre embryo, black arrowhead notes paucity of NCC in cardiac mesenchyme. FBA: first branchial arch, mandibular component. Data are representative of six embryos from each group.

Mentions: To analyze neural crest contributions to cardiac development in Spry1;Wnt1-Cre embryos, we examined histological sections taken from whole mount E9.5 Spry1:R26R;Wnt1-Cre embryos or R26R;Wnt-Cre littermates (Figure 10). Presumptive NCC marked by β-Gal staining in Figure 10A showed strong staining in the branchial arches and the outflow tract of control embryos. In contrast, Spry1;R26R;Wnt1-Cre embryos (Figure 10B) showed variable staining in the branchial arches and greatly diminished staining in the outflow tract. Histological sections (Figure 10C-F) revealed the first branchial arch was hypoplastic in the mutant vs. the control. The control embryos showed β-gal positive NCC colonizing the cardiac mesenchyme throughout the outflow tract and down into the bulbis cordis (Figure 10C, E). The mutant showed a paucity of β-gal positive neural crest cells colonizing the cardiac mesenchyme with a near total absence in the bulbis cordis. The outflow tract of the mutant was shortened and poorly rotated in comparison to the WT, which was elongated with a more spiral configuration. The lack of sufficient numbers of β-gal positive NCC colonizing the cardiac mesenchyme resulted in abnormal cardiac morphogenesis with the failure of the outflow tract to elongate normally, undergo normal cardiac looping, which as a consequence altered the rotation, alignment and septation of the outflow tract. Septation most likely did not occur due to the failure of the formation of the aorticopulmonary septum whose formation is critically dependent upon sufficient numbers of cardiac NCC colonizing and proliferating in the cardiac mesenchyme. DORV was a consequence of the malrotation and malalignment of the outflow tract, which was not positioned into its normal configuration between the atrioventricular valves. Cardiac NCC, in conjunction with the cells of the primary and secondary heart fields, are essential for normal formation of the endocardial cushions and conotruncal cushions. Deficiencies in these structures, which are dependant on cardiac NCC proliferation, signaling and interaction with the primary and secondary heart field cells, most likely led to the observed cardiac defects.


Conditional expression of Spry1 in neural crest causes craniofacial and cardiac defects.

Yang X, Kilgallen S, Andreeva V, Spicer DB, Pinz I, Friesel R - BMC Dev. Biol. (2010)

Spry1;Wnt1-Cre embryos show outflow tract defects at E9.5. (A, B) Whole mount β-gal staining of Cre-negative littermate control and Spry1;Wnt1-Cre E9.5 embryos. (A) WT shows intense β-gal staining of the outflow tract (white arrow) and branchial arches, indicating cells of NC origin. (B) Spry1;Wnt1-Cre E9.5 embryo, shows variable β-gal staining of the pharyngeal arches and reduced staining of the outflow tract (white arrowhead). (C, D) Sagittal sections of whole mount embryos with nuclear fast red counter-staining. (C) WT shows normal distribution of cardiac NCC within the outflow tract, with β-galactosidase positive NCC cells extending down to the bulbis cordis. Panel D, Spry1;Wnt1-Cre reveals outflow tract with reduced β-galactosidase positive NCC cells. In addition the first branchial arch, mandibular component, is greatly reduced in size relative to the WT. The outflow tract in Spry1;Wnt1-Cre embryos is shortened and does not adopt the spiral configuration as seen in the WT. (E,F) High power images of C,D; white boxed areas indicate field of view. (E) WT, black arrow indicates cardiac NCC contributing cardiac mesenchyme. (F) Spry1;Wnt1-Cre embryo, black arrowhead notes paucity of NCC in cardiac mesenchyme. FBA: first branchial arch, mandibular component. Data are representative of six embryos from each group.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC2874773&req=5

Figure 10: Spry1;Wnt1-Cre embryos show outflow tract defects at E9.5. (A, B) Whole mount β-gal staining of Cre-negative littermate control and Spry1;Wnt1-Cre E9.5 embryos. (A) WT shows intense β-gal staining of the outflow tract (white arrow) and branchial arches, indicating cells of NC origin. (B) Spry1;Wnt1-Cre E9.5 embryo, shows variable β-gal staining of the pharyngeal arches and reduced staining of the outflow tract (white arrowhead). (C, D) Sagittal sections of whole mount embryos with nuclear fast red counter-staining. (C) WT shows normal distribution of cardiac NCC within the outflow tract, with β-galactosidase positive NCC cells extending down to the bulbis cordis. Panel D, Spry1;Wnt1-Cre reveals outflow tract with reduced β-galactosidase positive NCC cells. In addition the first branchial arch, mandibular component, is greatly reduced in size relative to the WT. The outflow tract in Spry1;Wnt1-Cre embryos is shortened and does not adopt the spiral configuration as seen in the WT. (E,F) High power images of C,D; white boxed areas indicate field of view. (E) WT, black arrow indicates cardiac NCC contributing cardiac mesenchyme. (F) Spry1;Wnt1-Cre embryo, black arrowhead notes paucity of NCC in cardiac mesenchyme. FBA: first branchial arch, mandibular component. Data are representative of six embryos from each group.
Mentions: To analyze neural crest contributions to cardiac development in Spry1;Wnt1-Cre embryos, we examined histological sections taken from whole mount E9.5 Spry1:R26R;Wnt1-Cre embryos or R26R;Wnt-Cre littermates (Figure 10). Presumptive NCC marked by β-Gal staining in Figure 10A showed strong staining in the branchial arches and the outflow tract of control embryos. In contrast, Spry1;R26R;Wnt1-Cre embryos (Figure 10B) showed variable staining in the branchial arches and greatly diminished staining in the outflow tract. Histological sections (Figure 10C-F) revealed the first branchial arch was hypoplastic in the mutant vs. the control. The control embryos showed β-gal positive NCC colonizing the cardiac mesenchyme throughout the outflow tract and down into the bulbis cordis (Figure 10C, E). The mutant showed a paucity of β-gal positive neural crest cells colonizing the cardiac mesenchyme with a near total absence in the bulbis cordis. The outflow tract of the mutant was shortened and poorly rotated in comparison to the WT, which was elongated with a more spiral configuration. The lack of sufficient numbers of β-gal positive NCC colonizing the cardiac mesenchyme resulted in abnormal cardiac morphogenesis with the failure of the outflow tract to elongate normally, undergo normal cardiac looping, which as a consequence altered the rotation, alignment and septation of the outflow tract. Septation most likely did not occur due to the failure of the formation of the aorticopulmonary septum whose formation is critically dependent upon sufficient numbers of cardiac NCC colonizing and proliferating in the cardiac mesenchyme. DORV was a consequence of the malrotation and malalignment of the outflow tract, which was not positioned into its normal configuration between the atrioventricular valves. Cardiac NCC, in conjunction with the cells of the primary and secondary heart fields, are essential for normal formation of the endocardial cushions and conotruncal cushions. Deficiencies in these structures, which are dependant on cardiac NCC proliferation, signaling and interaction with the primary and secondary heart field cells, most likely led to the observed cardiac defects.

Bottom Line: Spry1;Wnt1-Cre embryos die perinatally and exhibit facial clefting, cleft palate, cardiac and cranial nerve defects.These defects appear to be the result of decreased proliferation and increased apoptosis of neural crest and neural crest-derived cell populations.In addition, the domains of expression of several key transcription factors important to normal craniofacial and cardiac development including AP2, Msx2, Dlx5, and Dlx6 were reduced in Spry1;Wnt1-Cre transgenic embryos.

View Article: PubMed Central - HTML - PubMed

Affiliation: Center for Molecular Medicine, Maine Medical Center Research Institute, Scarborough, ME 04074, USA.

ABSTRACT

Background: Growth factors and their receptors are mediators of organogenesis and must be tightly regulated in a temporal and spatial manner for proper tissue morphogenesis. Intracellular regulators of growth factor signaling pathways provide an additional level of control. Members of the Sprouty family negatively regulate receptor tyrosine kinase pathways in several developmental contexts. To gain insight into the role of Spry1 in neural crest development, we analyzed the developmental effects of conditional expression of Spry1 in neural crest-derived tissues.

Results: Here we report that conditional expression of Spry1 in neural crest cells causes defects in craniofacial and cardiac development in mice. Spry1;Wnt1-Cre embryos die perinatally and exhibit facial clefting, cleft palate, cardiac and cranial nerve defects. These defects appear to be the result of decreased proliferation and increased apoptosis of neural crest and neural crest-derived cell populations. In addition, the domains of expression of several key transcription factors important to normal craniofacial and cardiac development including AP2, Msx2, Dlx5, and Dlx6 were reduced in Spry1;Wnt1-Cre transgenic embryos.

Conclusion: Collectively, these data suggest that Spry1 is an important regulator of craniofacial and cardiac morphogenesis and perturbations in Spry1 levels may contribute to congenital disorders involving tissues of neural crest origin.

Show MeSH
Related in: MedlinePlus