Notch2 contributes to the development of aortic and pulmonary valves in mouse embryonic heart

doi:10.12307/2023.059

Abstract

Abstract: BACKGROUND: Abnormalities of the arterial valve leaflets explain part of congenital heart malformations. Among of these, bicuspid aortic valve are the commonest congenital malformations in humans. However, the development process of the aortic and pulmonary valves in embryonic heart remains to be further elucidated.
OBJECTIVE: To observe the septation of outflow tract and the formation and remodeling of aortic and pulmonary valve anlagen in mouse embryonic heart.
METHODS: Healthy ICR mice of 2 months old, SPF grade including 16 females and 10 males, were caged at 18:00 every night at a ratio of 2:1 between females and males. The mice which found vaginal plugs at 6:00 the next day were recorded to be at 0.5 days of pregnancy. The mouse embryos at 9.5-15 days of embryonic age were selected for preparing paraffin-embedded serial sections. Immunohistochemical staining was used to observe the protein expression of Isl1, Nkx2.5, myosin heavy chain, Ap2α, Sox9, α-smooth muscle actin, Snail, Jag1, Notch2, Alk3, and p-Smad1/5/8 in the mouse embryonic heart at 9.5-15 days of embryonic age. Three-dimensional reconstruction of the heart was performed to observe changes in the outflow tract endocardial cushion and intercalated cushions as well as valve morphology.
RESULTS AND CONCLUSION: During embryonic days 9.5-15, endothelial cells in the outflow tract were transformed into mesenchymal cells in the cardiac jelly through epithelial-mesenchymal transformation, which participated in the formation of endocardial cushion in the outflow tract. From embryonic days 11 to 11.5, Isl1 positive cells in the pharyngeal mesenchyme extended into the outflow tract wall to form intercalated cushions, and epithelial-mesenchymal transformation was not observed during this process. Notch2 signal was involved in the formation of endocardial cushion in the outflow tract, but not in the formation of intercalated cushions. At embryonic day 12.5, the distal poles of the bilateral endocardial cushions of the outflow tract were fused to form the left and right coronary valve anlagen of the aorta and the left and right semilunar valve anlagen of the pulmonary trunk. The intercalated cushions formed the non-coronal valve anlagen of the aorta and the anterior semilunar valve anlagen of the pulmonary artery. The Notch2 signal was expressed in all valve anlagen. During embryonic days 13-15, the arterial valve anlagen were remodeled to form a slender valve and the expression of Notch2 signal in the valves gradually decreased. The results suggest that the mesenchymal cells in the intercalated cushions originate from the direct differentiation of Isl1 positive cells of the second heart field. Notch2 may not be involved in the formation of intercalated cushions, but may contribute to the formation and fusion of the outflow tract endocardial cushion and remodeling of the arterial valve anlagen.

Key words: mouse embryo, arterial valve, outflow tract endocardial cushion, epithelial-mesenchymal transformation, Notch2, secondary heart field, heart

CLC Number:

Wang Jiajia, Xie Jianshan, Li Hairong, Jing Yixin, Yang Yanping. Notch2 contributes to the development of aortic and pulmonary valves in mouse embryonic heart[J]. Chinese Journal of Tissue Engineering Research, 2023, 27(14): 2176-2181.

Figures/Tables 5

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