European Journal of Anatomy

Official Journal of The Spanish Society of Anatomy
Cover Volume 24 - Number 3
Eur J Anat, 24 (3): 211-220 (2020)

Late administration of caffeine affects cardiac maturation in chick embryos: a combined two and threedimensional morphogenetic and gene analyses

Nichapha Chandee1,2, Nutmethee Kruepunga1, Thanyarat Leckchaoum1, Pakpoom Thintharua1, Wattana Weerachatyanukul1, Somluk Asuvapongpatana1

1Department of Anatomy, Faculty of Science, Mahidol University, Bangkok, Thailand, 2School of Allied Health Science, Walailak University, Nakhon Si Thammarat, Thailand

ABSTRACT 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¬ģ 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.

Keywords: Chick embryo ‚?? Cardiac development ‚?? Cardiac maturation ‚?? Ventricular defect ‚?? Caffeine ‚?? 3D analysis

European Journal of anatomy
ISSN 2340-311X (Online)