In situ hybridisation of chick embryos with p53-specific probe and their immunostaining with anti-p53 antibodies

Abstract 

Tumor-suppressor protein p53 is an important regulator of cell cycle and apoptosis. On the level of embryo extracts it has been shown earlier that both p53 protein and mRNA are expressed in developing chicken. Here we describe the expression patterns of p53 mRNA and protein in developing chicken embryos (stages 2–12) using in situ hybridisation and immunostaining with p53-specific monoclonal antibody Mab421. p53 mRNA is equally localised all over the embryo in the stages observed. According to electron microscopy data a subfraction of p53 mRNA is bound to dissolving yolk granules expressing acid phosphatase activity characteristic for lysosomes. Protein p53 is synthesised starting from the medium primitive streak stage (stage 3) and reaches its maximum level at the full primitive streak stage. During these stages protein p53 is distributed evenly across the embryos. After gastrulation p53 protein remains visible at higher levels only in certain anlages and areas. In developing nervous system the expression is observable in neuroectoderm, during the closure of the neural tube and in mesenchyme in the area of migrating neural crest cells. In cardiogenesis protein p53 is expressed during formation of tubular heart in the epimyocardium, endocardium and cardiac jelly. p53 protein localises in the neurocoele (obviously connected with cellular debris) and cardiac jelly. Our data support the role of p53 in early development, especially during embryo gastrulation, the development of central nervous system, neural crest and heart. In some cases increased p53 amounts colocalise with the areas of intensive epithelium-mesenchyme transition.