Alteration of normal protein kinase C (PKC) function by environmental Pb exposure during neurodevelopment is hypothesized to be an important mechanism of toxicity underlying neurologic impairment. Previous studies have reported widely varying effects of Pb on PKC, possibly in part because of differences in in vitro and in vivo models used in those studies. Therefore, we tested the hypothesis that, with comparable tissue Pb levels, the effects of in vitro Pb exposure on brain PKC are the same as the effects caused by in vivo Pb exposure of intact animals. For chronic in vivo Pb exposure, female Long-Evans rats were exposed to Pb or vehicle from postnatal days 1 to 34-36 (n=10/treatment). For in vitro Pb exposure, homogenate of the frontal cortex region was exposed directly to Pb in an amount comparable to that accumulated in brain during chronic in vivo Pb exposure. Brain Pb levels were measured using ultraclean techniques and inductively coupled plasma mass spectrometry. PKC activity was subsequently determined in cytosolic and membrane subcellular fractions in the frontal cortex, hippocampus, and remaining brain regions. Results indicate that brain Pb levels following in vivo Pb exposure were increased approximately 20-fold above those of nonexposed animals (vehicle group [Pb] approximately 130ng Pb/g dry wt.). However, in vivo Pb exposure did not measurably alter brain PKC activity in the regions tested. In contrast, in vitro Pb exposure significantly increased PKC activity by approximately 20% in the frontal cortex homogenate membrane subcellular fraction. These results indicate that Pb added in vitro caused more dramatic effects than those produced by a comparable amount of Pb in the tissue from in vivo exposure. While the mechanisms underlying these outcomes are not clear, they suggest that in vitro models might not accurately reflect effects of chronic low-level in vivo Pb exposure.