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Cardiovascular Research 1995 30(2):262-269; doi:10.1016/S0008-6363(95)00036-4
© 1995 by European Society of Cardiology
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Copyright © 1995, European Society of Cardiology

Immunohistochemical evaluation of the developing outflow tract in the rat: achieving aortic to mitral fibrous continuity

M. Jackson*,a, M.G. Connella, A. Smitha and R.H. Andersonb

aThe Institute of Child Health, University of Liverpool, Royal Liverpool Children's Hospital Alder Hey, Eaton Road, Liverpool, L12 2AP, UK
bRoyal Brompton National Heart and Lung Institute, Dovehouse Street, London, SW3 6LY, UK

* Corresponding author. Tel. (+51) 228 4811, ext. 2438.

Objectives: To study the development of aortic to mitral fibrous continuity in the normal rat heart. Methods: The hearts and great vessels of normally developed rat embryos and fetuses aged between 13.25 and 19.75 days of gestation were studied in conjunction with those of newborns aged 2 and 7 days post-partum. Standard histological methods and monoclonal antibodies raised against alpha smooth muscle actin (clone 1A4) and ventricular beta myosin heavy chain were used to demonstrate the ventricular outlets, ventriculo-arterial junction, inner heart curve and aortic infundibulum from the early stages of aortopulmonary septation to attainment of their definitive morphology. Results: The two antibodies demonstrated temporal specificity (actin specificity increased post-partum; myosin specificity maximal during fetal period) in the labelling of their intended structures which correlated with their known developmental profile. Full-thickness fibrous continuity between aortic and mitral valves was not complete until 1 week after birth. After ventricular septation was complete, and thereafter towards the end of fetal life and beyond, separation was maintained by a muscular structure histologically identical to the vestigial netro-aortic root branch of the conduction tissue, a structure known to be derived from the primitive ventricular myocardium within the environs of the inner heart curve. Conclusions: Ventricular septation (occurring relatively early) and the attainment of fibrous continuity (occurring relatively late in development) are two independent processes. Muscular tissue separating left-sided arterial and atrioventricular valves is not derived from the aortic infundibulum but from the inner heart curve. Persistence of this structure is a feature of normal rat heart development and needs to be recognised when working with rodent-based animal models of congenital heart disease aimed at studying the disruption of the development of the ventricular outflow tracts.

KEYWORDS Fibrous continuity; Development; Rat heart; Congenital heart disease; Immunohistochemistry


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