Regulation of A-currents by cell-cell interactions and neurotrophic factors in developing chick parasympathetic neurones.

1. The developmental regulation of ion channel expression was studied in parasympathetic neurones isolated from the chick ciliary ganglion. Whole-cell patch clamp recordings were made from ciliary ganglion neurones that were removed from the embryo on the ninth embryonic day (E9) and maintained in dissociated cell culture for an additional 4 days. Previous studies have shown that the expression of a transient voltage-activated K+ current (IA) is regulated by unidentified environmental stimuli during these developmental stages. 2. The effect of interactions between neurones and target tissue on the expression of IA was tested by co-culturing ciliary ganglion neurones with chick striated muscle cells. Neurones from the nerve-muscle co-cultures expressed normal amplitudes of IA, but the neurones did not express normal levels of IA when they were plated onto lysed muscle fibres. 3. The effect of interactions between ganglionic neurones and non-neuronal ganglionic cells was tested by culturing ganglia as explants rather than as dissociated cells. Neurones isolated from the explant cultures did not express normal levels of IA. Similarly, when dissociated ganglionic neurones were co-cultured with fibroblasts isolated from embryonic chick skin, they did not express normal amplitudes of IA. 4. Chronic depolarization caused by growing ciliary ganglion neurones in the presence of elevated K+ concentrations did not allow for the normal expression of IA, although it did promote the survival of these neurones in vitro. 5. Addition of 40 ng ml-1 of recombinant human ciliary neurotrophic factor (CNTF) or basic fibroblast growth factor (bFGF) to the cell culture medium had no effect on IA expression in developing chick ciliary ganglion neurones. However, 40 ng ml-1 of acidic fibroblast growth factor (aFGF) stimulated the expression of IA. All trophic factors promoted the growth and survival of ciliary ganglion neurones in vitro. 6. Dissociated ciliary ganglion neurones were maintained in a culture medium containing an aqueous extract of the whole brain. Neurones developing under these conditions expressed normal levels of IA. The stimulatory activity of the brain extract was destroyed by heating. 7. The expression of IA in chick ciliary ganglion neurones developing in vitro can be regulated by soluble growth factors and by interactions with certain other cell types. Similar interactions may regulate the expression of IA in ciliary ganglion neurones developing in situ.