It is important to determine how the signaling pathways for the short-term effects of nicotine (catecholamine secretion, phosphorylation of tyrosine hydroxylase) differ from those required for changes in gene expression. Our aim was to distinguish the pathways involved in short- and long-term nicotinic stimulation. PC12 cells were treated with several concentrations of nicotine from 10 micro M to 1 mM. All elicited a rapid and transient rise in [Ca(2+)](i), which was concentration dependent. After several minutes of continued exposure, a second smaller sustained rise in [Ca(2+)](i) was only observed with intermediate concentrations of nicotine (50-200 micro M). This sustained rise was not observed in cells pretreated with alpha-bungarotoxin (alpha-BTX). alpha-BTX also prevented the elevation of tyrosine hydroxylase mRNA by nicotine. The effects of brief and prolonged treatment with nicotine on the signaling pathways involved in changes in [Ca(2+)](i) and induction of tyrosine hydroxylase gene expression are summarized. The results indicate that nicotine may elicit different signaling pathways depending on the concentration. The sustained elevation of [Ca(2+)](i) via activation of alpha7 nAChRs is proposed as the mechanism leading to increased tyrosine hydroxylase gene expression.