Sprouting of neurites by sympathetic neurons from newborn rats was studied in compartmentalized cultures. The neuronal cell bodies resided in proximal compartments, and neurites penetrated silicone grease barriers and elongated within distal compartments. Nerve growth factor (NGF) was initially supplied at 1 microgram/ml in all compartments, but was subsequently withdrawn from proximal compartments and for a time was only supplied to distal neurites. Little or no neurite growth was observed in proximal compartments after NGF withdrawal, but reintroduction of NGF resulted in substantial neurite growth over the next few days which was shown to have originated from local sprouting within the proximal compartments. This result is distinct from previous work on NGF-enhanced nerve fiber elongation in demonstrating that quiescent, NGF-deprived regions of sympathetic neurons sprout neurites in response to local reexposure to NGF.