Differentiation of the chick embryo floor plate

Abstract

In a number of species, the floor plate of the developing neural tube and spinal cord has been ascribed specialized functions associated with the patterning of neuronal differentiation. The differentiation of the floor plate itself is believed to be closely related to the presence of the underlying notochord. Grafting experiments have previously shown that in the chick embryo an implanted segment of notochord is capable of inducing the adjacent host neural plate or neural tube to produce an additional floor plate, although the inductive effect diminishes with increasing age of the host. We have examined the potential of notochord to promote the appearance of floor plate-like structures from neural tube tissue in culture. To facilitate this, it was necessary initially to examine the immunoreactivity of the early neural tube and floor plate in situ and in vitro with a panel of antibodies to identify a suitable marker for floor plate differentiation in vitro. In situ, the differentiation of the floor plate was characterized by a lack of immunoperoxidase staining with antibody to neurofilaments and the monoclonal antibody HNK-1 throughout the period examined. This distinguished the floor plate from other regions of the neural tube, and was in contrast to its conspicuous affinity for antibodies to N-CAM and highly sialylated N-CAM, which also stained several closely adjacent regions of the neural tube over the period examined. We also found that oligodendrocytes occurred both in the floor plate and in the flanking ventral neural tube, and that astrocytes were too poorly represented throughout the neural tube at the stages examined to be useful markers of floor plate differentiation. We therefore concluded that only the anti-neurofilament and the HNK-1 antibodies were potentially useful markers for floor plate differentiation. When these antibodies were tested on cells in culture, neural tube tissue showed the presence of neurofilament- and HNK-1-positive neurites, while floor plate cultures showed few of these. These distributions were consistent with those demonstrated in situ. However, cells staining positively for N-CAM, sialylated N-CAM and the glial cell markers were relatively sparse in floor plate cultures, suggesting that these epitopes were not retained or were masked in cultured cells. As a result of these experiments, we selected the absence of neurofilament-positive cells as a marker for floor plate differentiation in culture.(ABSTRACT TRUNCATED AT 400 WORDS)