TY - JOUR A1 - , T1 - Late administration of caffeine affects cardiac maturation in chick embryos: a combined two and threedimensional morphogenetic and gene analyses JO - Eur. J. Anat. SN - 1136-4890 Y1 - 2020 VL - 24 SP - 211 EP - 220 UR - http://www.eurjanat.com/web/paper.php?id=190578sa KW - Chick embryo – Cardiac development – Cardiac maturation – Ventricular defect – Caffeine – 3D analysis N2 - Cardiac malformations are very prevalent and can be caused both by defective genes and envi-ronmental teratogens. Among the latter, caffeine causes malformations when exposed during early cardiac development, whereas its later effects are still unclear. We exposed three-day incubated (D3) chick embryos to 2 mg caffeine and analyzed them at D5, D7 and D9. The embryos were serially sec-tioned and analyzed two-dimensionally.Alternatively, the sections of D9 embryos were reconstructed three-dimensionally using Amira® software and analyzed volumetrically. The expres-sion of genes involved in endothelial-mesenchymal transformation (EMT) was studied by real-time PCR. Interestingly, caffeine treatment at D3 em-bryos did not induce cardiac malformations, but did delay growth, in particular that of the ventricles and ventricular trabeculae. Furthermore, it affected EMT in the endocardial cushion and atrioventricu-lar valves. Gene-expression analysis revealed that caffeine had a progressively deleterious effect on the expressions of GATA4, MMP2, SNAIL1, TWIST1, and VIMENTIN. The effect of late caf-feine administration on the chicken embryos would provide suggestive evident towards a possible heart developmental defect in humans, particularly heavy caffeine consumers during pregnancy. ER -