The functional expression of the Ca2+-activated K+ current (IK[Ca]) is dependent on cell-cell interactions in developing chick autonomic neurons. In chick ciliary ganglion (CG) neurons, expression of macroscopic IK[Ca] coincides with the formation of synapses with target tissues. CG neurons that develop in vivo in the absence of normal target tissues fail to express functional IK[Ca], although voltage-activated Ca2+ currents and most other ionic currents are expressed at normal amplitudes and densities. CG neurons placed in cell culture prior to formation of synapses with target tissues also fail to express macroscopic IK[Ca]. However, CG neurons cultured in the presence of a heat- and trypsin-sensitive extract of target tissues express IK[Ca] at normal levels. Similarly, interactions with target tissue appear to regulate the expression of whole-cell IK[Ca] in developing chick sympathetic ganglion neurons, although the relevant trophic factors appear to be different from those required by CG neurons. In addition to target tissue interactions, an intact preganglionic innervation is required for the normal in vivo development of IK[Ca] in chick CG neurons. The trophic effects of the afferent innervation do not require synaptic activation of the CG neurons, indicating secretion of a trophic factor, possibly an isoform of beta-neuregulin. The results are consistent with the hypothesis that target- and nerve terminal-derived trophic factors interact at a posttranslational level in the regulation of a functional IK[Ca]. Together, this body of data demonstrates an essential role for cell-cell interactions in the differentiation of neuronal excitability.