Tissues exhibiting inhibitory [correction of inhibiory] and repulsive activities during the initial stages of neurite outgrowth from the dorsal root ganglion in the chick embryo

To elucidate the mechanisms underlying the projection of dorsal root ganglion (DRG) axons into the dorsal root entry zone in the dorsolateral region of the spinal cord, we examined tissue interactions which affect neurite outgrowth from DRG. We cultured explants or dissociated cells of DRG from embryonic day 4 (E4) chick embryos in combination with E3 spinal cord, notochord, and dermomyotome in three-dimensional collagen gels. The ventral spinal cord, notochord, and dermomyotome, which are located close to the initial projection pathway of DRG but do not receive direct innervation, strongly inhibited DRG neurite outgrowth and repelled DRG neurites. These inhibitory/repulsive cues appear diffusible in nature, because this activity was observed in the absence of direct contacts between tissue explants and DRG neurites. Furthermore, in heterochronic cultures, E9 DRG lost its responsiveness to inhibitory/repulsive factors from E3 ventral spinal cord, while retaining responsiveness to E3 notochord and dermomyotome, suggesting that the E3 ventral spinal cord may secrete a different inhibitory/repulsive signal than notochord and dermomyotome. Putative inhibitory/repulsive signals secreted from tissues along the axonal pathway may serve to guide growing DRG axons to the dorsal root entry zone.