Between stage B and C the notochord first arises.
In the hindermost sections of this stage the hypoblast retains a perfectly normal structure and uniform thickness throughout. In the next few sections (Pl. 10, fig. 1c, ch´) a slight thickening is to be observed in the hypoblast, immediately below the medullary canal. The layer, which elsewhere is composed of a single row of cells, here becomes two cells deep, but no sign of a division into two layers exhibited.
In the next few sections the thickening of the hypoblast becomes much more pronounced; we have, in fact, a ridge projecting from the hypoblast towards the epiblast (Pl. 10, fig. 1b, ch´).
This ridge is pressed firmly against the epiblast, and causes in it a slight indentation. The hypoblast in the region of the ridge is formed of two layers of cells, the ridge being entirely due to the uppermost of the two.
In sections in front of this a cylindrical rod, which can at once be recognised as the notochord and is continuous with the ridge just described, begins to be split off from the hypoblast. It is difficult to say at what point the separation of this rod from the hypoblast is completed, since all intermediate gradations between complete separation and complete attachment are to be seen.
Where the separation first appears, a fairly thick bridge of hypoblast is left connecting the two lateral halves of the layer, but anteriorly this bridge becomes excessively delicate and thin (Pl. 10, fig. 1a), and in some cases is barely visible except with high powers.
From the series of sections represented, it is clear that the notochord commences to be separated from the hypoblast anteriorly, and that the separation gradually extends backwards.
The posterior extremity of the notochord remains for a long time attached to the hypoblast; and it is not till the end of the period treated of in this chapter that it becomes completely free.
A sheath is formed around the notochord, very soon after its formation, at a stage intermediate between stages C and D. This sheath is very delicate, though it stains with both osmic acid and hæmatoxylin. I conclude from its subsequent history, that it is to be regarded as a product of the cells of the notochord, but at the same time it should be stated that it precisely resembles membrane-like structures, which I have already described as being probably artificial.
Towards the end of this period the cells of the notochord become very much flattened vertically, and cause the well-known stratified appearance which characterises the notochord in longitudinal sections. In transverse sections the outlines of the cells of the notochord appear rounded.