Neurotrophic factors released by target tissues maintain the survival and differentiation of innervating neurons. The manner by which these target-derived neurotrophic proteins communicate with innervating neurons has been actively pursued for over three decades. The present chapter describes a technique for preparing and maintaining compartmented chambers for culturing neurons derived from either superior cervical ganglia (sympathetic neurons) or dorsal root ganglia (sensory neurons). This system recapitulates the selective stimulation of neuron terminals that occurs in vivo following release of target-derived neurotrophins.