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Review
. 2011 Oct;21(5):573-8.
doi: 10.1016/j.gde.2011.07.004. Epub 2011 Sep 4.

Imaging heart development using high-resolution episcopic microscopy

Timothy J Mohun  1 , Wolfgang J Weninger
Affiliations
Review

Imaging heart development using high-resolution episcopic microscopy

Timothy J Mohun et al. Curr Opin Genet Dev. 2011 Oct.

Abstract

Development of the heart in vertebrate embryos is a complex process in which the organ is continually remodelled as chambers are formed, valves sculpted and connections established to the developing vascular system. Investigating the genetic programmes driving these changes and the environmental factors that may influence them is critical for our understanding of congenital heart disease. A recurrent challenge in this work is how to integrate studies as diverse as those of cardiac gene function and regulation with an appreciation of the localised interactions between cardiac tissues not to mention the manner in which both may be affected by cardiac function itself. Meeting this challenge requires an accurate way to analyse the changes in 3D morphology of the developing heart, which can be swift or protracted and both dramatic or subtle in consequence. Here we review the use of high-resolution episcopic microscopy as a simple and effective means to examine organ structure and one that allows modern computing methods pioneered by clinical imaging to be applied to the embryonic heart.

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Figures

Figure 1
Figure 1
(a) An HREM image taken from an isolated chick embryo heart (HH stage 32) at the level of the developing atrioventricular junction, showing the range of grey levels associated with different tissue types. (b) 3D models of mouse embryo hearts isolated at E14.5 (when chamber septation is just completed) and E18.5 (shortly before birth) with that of the adult mouse. Models (not to scale) are eroded along a transverse plane from aortic valve to ventricular apex. This graphically illustrates the change in ventricular wall thickness and the mesh of spongy trabeculation that accompanies heart development.
Figure 2
Figure 2
Measuring the diameters of the great intrathoracal arteries of a 14.5 dpc mouse foetus. (a) Great intrathoracic arteries (red) in situ. Surface rendered 3D models of the arteries are displayed together with the original HREM section plane and two re-section planes cutting perpendicular to each other and perpendicular to the original section plane through the HREM volume data. (b) 3D surface model and two oblique re-section planes. Note that the virtual planes were orientated to cut perpendicular to the longitudinal axis of the respective blood vessel segment through the original volume data. (c) and (d) Virtual section planes shown in (b). aa = ascending aorta, bt = brachiocephalic trunk, lc = left common carotid artery, ls = left subclavian artery, da = descending aorta, pt = pulmonary trunk, pa = pulmonary artery, db = ductus arteriosus (Botalli), av = aortic valve, pv = pulmonary valve, at = atrium, cv = cardiac ventricle, li = liver, r = rib, cl = clavicle, ve = forming vertebrae sc = spinal chord, sg = signal ganglion, t = trachea, e = oesophagus. Scale bar = 200 μm.
Figure 3
Figure 3
(a) 3D models of β-galactosidase expression in an E11.5 Islet1-lacZ embryo, captured by dual wavelength HREM and pseudo-coloured magenta in the whole embryo (left) or white in the heart (right). The latter is eroded in the transverse plane to show expression in the central pharyngeal region (including the dorsal roof of the aortic sac) as well as both proximal and distal walls of the outflow tract. (b) Images obtained by ‘virtual endoscopy’ using HREM data from an E18.5 mouse embryo heart, revealing remarkable details of heart structure. The left panel shows a view of the right atrium from the entrance of the right superior caval vein. Note the smooth ventral floor of the right atrium surrounding the coronary sinus, flanked on one side by the trabeculae of the right atrial appendage and edge of the tricuspid valve on the other. The right panel shows a view of the trabecular lattice within the right atrium, viewed through the tricuspid valve.
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