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Fig. 1. Hhex–/– mice have multiple cardiovascular abnormalities. (A,B) 13.5 embryos and (C,D) whole hearts. (A,B) Compared to Hhex+/+ embryos, Hhex–/– embryos have no livers (asterisks), forebrain truncations (arrowheads), and markedly dilated blood vessels (arrows in B). Hearts in Hhex–/– mice at E13.5 have a small right ventricle (arrows in C and D). (E-P) Transverse sections of E13.5 embryos. (E) Section of Hhex+/+ heart inferior to outflow tracts showing normal morphology. (F) Hhex–/– heart with hypoplastic RV (arrow) and abnormal accumulation of ECCs (box). (G) Higher power view of boxed region in F reveals abnormal accumulation of cells in the atrio-ventricular cushion (AVC) region that are mesenchymal in morphology and represent ECCs. (H,I) RV outflow tract. (H) The Hhex+/+ heart has a small region of ECCs just inferior to the pulmonic valve (asterisk). The arrows indicate the region of the RV outflow tract that is comprised of ECCs. (I) The Hhex–/– heart shows a marked increase in ECCs with subsequent narrowing of RV outflow tract. Arrows indicate the region of RVOT that is comprised of ECCs and the asterisk denotes the pulmonic valve. (J) A more caudal section of the same heart as in I showing the aortic valve (asterisk) communicating with RV, indicating the presence of DORV. (K,L) AV valve region showing abnormal accumulation of ECCs and dysplastic AV valves in Hhex–/– mice compared to wild-type mice. (M-Q) Ventricles of Hhex+/+ and Hhex–/– hearts. In the Hhex–/– heart, a VSD (double arrow) is present and the myocardium is abnormally thin (O). High power view of boxed areas in (M) and (O) is shown in (N) and (P) and highlights the abnormally thin compact myocardial layer in Hhex–/– hearts. The thickness of the compact layer of the myocardium is indicated. Arrowheads point to epicardium. (Q) High power image through the myocardium of a severely affected embryo in which it is difficult to identify the presence any compact layer. RV, right ventricle; Ao, aorta; TV, tricuspid valve; MV, mitral valve; ECC, endocardial cushion cells; DORV, double outlet right ventricle; CL, compact layer; TL, trabecular layer.





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