Induction of neuronal differentiation by planar signals in Xenopus embryos

The induction of the central nervous system in amphibian embryos is mediated both by early planar signals produced by mesoderm at the dorsal lip and later vertical signals emanating from the dorsal mesoderm after involution. We have examined the role and spatial extent of planar signals in the induction of neuronal differentiation. Planar explants that included only the deep layer of the dorsal marginal zone, comprising both the dorsal mesoderm and the contiguous dorsal ectoderm, were isolated at the beginning of gastrulation. After removal of the epithelial layer, explants were maintained in modified Danilchik's medium until mid-neurula stages, when they were transferred to modified Danilchik's medium + 0.1% bovine serum albumin and cultured on laminin. Neurite outgrowth occurred in 90% of these planar explants. In contrast, little or no neuronal differentiation occurred in either ventral planar explants or explants of ectoderm alone. Video analysis of cell movements shows that large-scale cell mixing does not occur between mesoderm cells and ectoderm cells in planar explants. Retrograde labelling of neuronal cell bodies indicates that cells throughout the ectoderm undergo neuronal differentiation; neurons also differentiate in cultures of distal ectoderm isolated at early neurula stages from planar explants prepared at the beginning of gastrulation. These observations indicate that planar signals act over an extended range to induce neuronal differentiation. The inductive capacity of vertical signals was examined by recombining animal caps from ultra-violet (UV) irradiated embryos with involuted mesoderm from normal midgastrula embryos. Differentiation of either neurons or anterior neural structures occurred in 73% of vertical recombinates. Our results demonstrate that planar signals from the dorsal lip of the blastopore are capable of inducing neuronal differentiation over a considerable distance in the absence of epithelial confinement, convergence and extension, and mixing between the mesoderm and ectoderm.