We have previously shown that chronic antagonism of metabotropic glutamate receptors in the brain attenuates naloxone-precipitated withdrawal symptoms in rats treated chronically with subcutaneous (s.c.) morphine. Several subtypes of metabotropic glutamate receptors are directly linked, through a guanine nucleotide regulatory protein, to the phosphatidylinositol (p.i.) second messenger system. In the present investigation, we assessed the effect of inhibiting the products of p.i. hydrolysis on the development of opioid dependence. Thus, concurrently with subcutaneous morphine, we infused intracerebroventricularly (i.c.v.) in rats, various doses of chelerythrine, which selectively inhibits the activation of protein kinase C, and thapsigargin, which inhibits the release of intracellular Ca2+ when given chronically. Both chelerythrine and thapsigargin reduced the severity of naloxone-precipitated abstinence symptoms when infused i.c.v. at a dose of 10 nmol/day. A single injection of either chelerythrine or thapsigargin immediately prior to the precipitation of withdrawal failed to decrease the severity of abstinence symptoms. Our results suggest that by chronically inhibiting activity of the phosphatidylinositol system, the development of morphine dependence can be attenuated.