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Figure 5


Fig. 5. Antagonism of FGF signaling by overexpression of Spry2 disrupts OFT development. (A-I) Isolated E18.5 mouse hearts from controls (A-C) and Spry2-GOF;Mesp1Cre mutants (D-I). (A) Frontal view. (B) Superior view, ventral surface at top; the AV is dorsal and right of the PV. trv, tricuspid valve; mv, mitral valve. (C) Frontal view; RV wall removed to show PV and ventricular septum (VS). (D) Frontal view of type III PTA. (E) Superior view of embryo in D. (F) Frontal view of membranous VSD (arrowhead). (G) Superior view showing a bicuspid aortic valve (BAV, arrowhead). (H) Superior view of mutant with DORV; probe (black line) passes abnormally from the Ao into the RV. (I) Superior view, atria removed, of atrioventricular septal defect (AVSD) with incomplete fusion of the AVC cushions. (J-O) mRNA in situ hybridizations of E9.5 control (J,L,N) and Spry2-GOF;Mesp1Cre mutant (K,M,O) embryos. Mutant OFTs are short and aberrantly angulated (white lines) and the RV is hypoplastic. (J,K) Wnt11 transcripts are decreased in the mutant OFT. (L,M) Bmp4 transcripts are decreased in the mutant OFT (arrowhead) and in the SHF (arrow). (N,O) Crabp1 expression is decreased in mutant pharyngeal arch neural crest (arrowheads). (P,Q) Sections of E10.5 control (P) and Spry2-GOF;Mesp1Cre mutant (Q). Arrowhead points to reduced CNC and arrow to proximal cushion defects in the mutant. (R,S) Explant cultures of control (R) and Spry2-GOF;Mesp1Cre (S) OFTs show reduction in EMT when reception of FGF signaling in mesodermal cells is compromised. (T,U) Immunohistochemical detection of phosphohistone H3 (pHH3, red) and Isl1 (green) in E8.5 [11-somite stage (ss)] embryos. (V) Quantitation of pHH3+ cells as a percentge of the total number of cells in the SHF and OFT of E8.5 (10-11ss) control and Spry2-GOF;Mesp1Cre mutants. Proliferation is significantly decreased in both tissues (SHF, P=0.039; OFT, P=0.05). Three sections per embryo were counted and n=3 embryos per genotype. The s.d. was measured with Student's t-test.





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