DYNIMHEART

Defining the Biomechanics of the Developing Heart through High-Speed Dynamic Fluorescent Imaging in Transgenic Quail Embryos

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 Obiettivo del progetto (Objective)

'Vertebrate heart morphogenesis is a complex process that integrates different structures and cell types compelled to interact by genetic and epigenetic factors. Mistakes at any step from cell-commitment to valve formation will have a major impact on heart morphogenesis leading to congenital heart disease. Avian embryos offer an ideal system to study heart development because the development of the four-chambered avian heart is comparable to that of the four-chambered mammalian heart. The avian embryo is readily amenable for optical accessibility and permits direct observation of the cellular movements comprising heart formation in a warm-blooded experimental system. We use quail because of their small sized eggs, their moderately sized breeding adults, their short generation time, and their transgenic feasibility. The transgenic quails developed in the hosting laboratory expressing fluorescent proteins constitute a new model syste. Bright trangenic quails improve the ability to dynamically image vascular development during embryogenesis. Our proposal aims to study the formation of the heart during the embryonic development of quails with special attention to fate mapping myocardium and endocardium progenitors. We will carry out dynamic imaging, using laser scanning microscopes, to image live transgenic or chimeric quail lines that express fluorescent proteins ubiquitously, in the endocardium and in additional tissues or subcellular structures. Using a high-speed confocal microscope we aim to capture the heartbeat at different z-depths. We can reconstruct the data into a 4D figure allowing us to conclude the impedance action of the heart on the endocardium wall.'

Introduzione (Teaser)

Embryogenesis is a complex process where a few cells differentiate to give rise to a whole embryo. Identifying the triggers and determinants in each step should help us comprehend why sometimes things go wrong.

Descrizione progetto (Article)

Formation of the vertebrate heart during development is an intricate process involving the complex interaction of genetic and epigenetic factors. From commitment to cardiac cell fate to valve formation, it has to be carefully and infallibly regulated. Mistakes in any step of the process could lead to congenital heart disorders.

Avian heart morphogenesis resembles the mammalian process, and the avian and mammalian heart share a similar four-chamber architecture. As a result, the avian embryo represents a suitable model for studying heart development and its optical accessibility permits direct observation of the cellular movements.

Scientists on the EU-funded DYNIMHEART project aim to further advance this system through dynamic fluorescent imaging of vascular development. To this end, they propose to generate transgenic quails that express fluorescent proteins in the endocardium or endothelial cells. Based on the spectral signatures of these cells, it will be possible to map their movement and distinguish them from other cell types. They have also developed specialised software, facilitating fluorescent cell tracking and together with high-speed confocal microscopy allows data to be reconstructed into 4D images.

Special emphasis has been given to activation of the Ephrin pathway, an essential component of tissue boundary formation. As an indicator for Ephrin activation, the consortium has used the phosphorylation of tyrosine kinase receptors. Detection of receptor aggregation in real time provides a further angle for studying cell-to-cell interactions during tissue morphogenesis.

Taken together, DYNIMHEART activities are anticipated to provide unprecedented insight into the spatiotemporal development of the heart. They should also unveil novel molecules with a fundamental role in cardiac morphogenesis.