To better understand the regulatory processes underlying axonal pathfinding we analyzed the embryonic expression of seven murine semaphorin genes by in situ hybridization In the spinal cord, transcripts of all seven semaphorin genes were detected from Embryonic Day 11.5 (E11.5) onward and restricted to distinct regions at E15.5. Interestingly, semE, F, and G mRNAs were in addition differentially expressed in the ventricular zone of the telencephalon. In order to correlate these expression patterns to the behavior of different types of sensory afferents, we tested their response to recombinant semaphorin proteins. Specific subpopulations of dorsal root ganglion sensory neurons displayed a developmentally regulated differential response to Sem D. Whereas extension of both NGF- and NT-3-dependent neurites was inhibited by Sem D at E12.5, only neurites formed in the presence of NGF responded at E14.5. This suggests that Sem D may be involved in preventing an early penetration of the spinal cord by sensory afferents and subsequently shaping their lamina-specific termination.