Differential in vitro degradation of particular Fos family members expressed by kainic acid in nuclear and cytosolic fractions of murine hippocampus

Mice were injected with kainic acid (KA) at a convulsive dose, followed by homogenization of the hippocampus in the presence of different protease and phosphatase inhibitors, and subsequent preparation of nuclear and cytosolic fractions. An intraperitoneal injection of KA resulted in marked expression of particular Fos family members, including c-Fos, Fra-2, and Fos-B, but not Fra-1 proteins, in both fractions 2 to 18 h after administration. These fractions were individually incubated at 30 degrees C for 1 to 18 h for determination of in vitro degradation. Similarly rapid degradation was seen with c-Fos protein between nuclear fractions obtained 2 and 18 h after administration, while no significant degradation was found for c-Fos protein in cytosolic fractions obtained 2 h after administration during incubation. By contrast, in vitro incubation led to rapid degradation of c-Fos protein in cytosolic fractions obtained 18 h after administration. Degradation profiles were peculiar to each member protein in nuclear and cytosolic fractions obtained 2 and 18 h after administration. Dialysis prevented degradation of c-Fos protein in nuclear fractions without markedly affecting that in cytosolic fractions in a manner independent of the time after administration. The addition of inhibitors for phosphatases, but not for proteases, accelerated the degradation of c-Fos protein in nuclear fractions previously dialyzed. These results suggest that in vivo KA signals may modulate heterologous machineries responsible for breakdown of each Fos family member in a unique manner in nuclear fractions, rather than cytosolic fractions, of murine hippocampus.