Mitogen-activated protein kinases (MAPK)–signaling pathways play key roles in cytoplasmic-nuclear signal transmission in response to various extracellular stimuli. In this study, we investigated the effect of repeated fasting stress on activation of the 3 members of the MAPK family, the extracellular signal-regulated kinase (ERK), the c-Jun NH2-terminal kinase (JNK), and the p38 mitogen-activated protein kinase (p38 kinase), in rat liver. Immunecomplex kinase assays showed that ERK and JNK were significantly activated in the liver extract from fasted rats whereas p38 kinase showed no activation. In an immunohistochemical study, the phosphorylated and activated form of ERK (p-ERK) was abundantly expressed in pericentral hepatocytes of fasted liver compared with those of the control. On the other hand, the phosphorylated and activated form of JNK (p-JNK) was highly expressed in irregular-shaped cells along the sinusoidal lining of fasted liver. A double immunofluorescent study to identify p-JNK immunoreactive cells revealed them to be Kupffer cells, which are the resident hepatic macrophages. In conclusion, ERK and JNK are selectively activated in distinct cell types of rat liver by repeated fasting stress.