Activation and autophosphorylation of apoptosis signal-regulating kinase 1 (ASK1) following cerebral ischemia in rat hippocampus.

We investigated the expression, activation and autophosphorylation of apoptosis signal-regulating kinase 1 (ASK1) in rat hippocampus after cerebral ischemia. The in vitro kinase assay showed that ASK1 activity gradually increased while the autophosphorylation of ASK1 gradually reduced during 5, 15 and 30 min of cerebral ischemia. At various time points of reperfusion, the activation and autophosphorylation of ASK1 reached a high point at 30 min and reduced to basal level at 6 h and then slightly increased at 3 d compared with sham operation. Both of the increases of ASK1 activation and autophosphorylation were suppressed by N-acetylcysteine, a well-known antioxidant, which was administered to the Sprague-Dawley rat 20 min before cerebral ischemia. Immunoprecipitation and Western blotting assay showed that there was no obvious change in the amount of ASK1 at each time point compared with sham control. Our results suggest that ASK1 protein which is known as an upstream mediator of JNK/p38 mitogen-actived protein kinase (MAPK) activation may play an important role in signal transduction in response to ischemic stress, given the fact that activation of JNK/p38 MAPK and subsequent phosphorylation of c-Jun are involved in the apoptotic pathway in cerebral ischemia. Copyright 2002 Elsevier Science Ireland Ltd.