Ultrastructure of the neural canal closure in the chicken embryo

We have studied the neuroectoderm of the chicken embryo, from the beginning of somitic segmentation up to the state at which it has seven somities, i.e. the period covering the passage from the 'neural canal' into the 'neural tube'. This paper was devoted mainly to the study of the ultrastructural cytodifferentiation which takes place during the stages in which the neural canal closes up, and at the level of the first area of contact between the 'neural crests'-roughly at the level of the third somite. We used eggs from a hen of the White Leghorn breed, incubated at 38 degrees C, from which we extracted chicken embryos after 24-30 h of incubation, corresponding to Hamburger-Hamilton's stages 7, 8 and 9. Thus we were able to obtain several series of embryos with three, six and seven somites. The neural canal, or tube, at the level of the third somite, was fixed in glutaraldehyde at 6.25% for 30 min and postfixed in 1% osmium tetroxide for 2 h embedded in Araldite, and the sections were then stained with lead citrate. We observed that the vacuoles in the free edge of the neural canal gradually disappear as the canal closes up, while we gradually witness the appearance of the 'closure apparatus' (or the safety or occlusion apparatus) of what is beginning to form the ependymal epithelium, and the first rudimentary outlines of the cilia. All these changes begin to be observed at the seven-somite stage, i.e. when the neural canal is beginning to close up. The 'closure apparatus' consists of a number of intercellular joint complexes, of the 'close-join't type, between which we observe a number of fine filaments, like a terminal velum', or veil, which we call 'interconnecting filaments'. In the 'raphe', whereby contact is established between the neural crests, we observe the initial stages of fusion between the vacuolated edges, with the plasmatic membrane of these cells forming very fine cytoplasmic 'tongues' which interdigitate with cells from the opposite neural crest and finally constitute the so-called close joints.