Prevention of PC12 Cell Death byN-Acetylcysteine Requires Activation of the Ras Pathway

Abstract

We have shown that N-acetylcysteine (NAC) promotes survival of sympathetic neurons and pheochromocytoma (PC12) cells in the absence of trophic factors. This action of NAC was not related to its antioxidant properties or ability to increase intracellular glutathione levels but was instead dependent on ongoing transcription and seemed attributable to the action of NAC as a reducing agent. Here, we investigate the mechanism by which NAC promotes neuronal survival. We show that NAC activates the Ras-extracellular signal-regulated kinase (ERK) pathway in PC12 cells. Ras activation by NAC seems necessary for survival in that it is unable to sustain serum-deprived PC12 MM17-26 cells constitutively expressing a dominantnegative form of Ras. Promotion of PC12 cell survival by NAC is totally blocked by PD98059, an inhibitor of the ERKactivating MAP kinase/ERK kinase, suggesting a required role for ERK activation in the NAC mechanism. In contrast, LY294002 and wortmannin, inhibitors of phosphatidylinositol 3-kinase (PI3K) that partially block NGF-promoted PC12 cell survival, have no effect on prevention of death by NAC. We hypothesized previously that the ability of NAC to promote survival correlates with its antiproliferative properties. However, although NAC does not protect PC12 MM17-26 cells from loss of trophic support, it does inhibit their capacity to synthesize DNA. Thus, the antiproliferative effect of NAC does not require Ras activation, and inhibition of DNA synthesis is insufficient to mediate NAC-promoted survival. These findings highlight the role of Ras-ERK activation in the mechanism by which NAC prevents neuronal death after loss of trophic support.

Tetrahydrobiopterin Enhances Apoptotic PC12 Cell Death following Withdrawal of Trophic Support

“…7A). Previous studies have demonstrated that the anti-apoptotic effects of NAC in PC12 cells are not due to its antioxidant capacity (60) but are instead dependent on new transcription and the activation of the Ras-ERK signaling pathway (61). Given our data with inhibitors of oxidative stress, we feel that NAC prevents BH 4 -induced DNA fragmentation by a mechanism that does not involve its antioxidant potential.…”

Early Single Cell Bifurcation of Pro- and Antiapoptotic States during Oxidative Stress

“…In PC-12 cells, ERK is mainly activated by growth factors and has been shown to be associated with cell proliferation, differentiation, and promotion of cell survival (13,16,58,59). H 2 O 2 increased the mRNA expression of the ERKdependent genes egr1, c-jun, c-fos, and mkp1.…”

The M1 Receptor Is Required for Muscarinic Activation of Mitogen-activated Protein (MAP) Kinase in Murine Cerebral Cortical Neurons

“…The MAP kinases ERK1 and ERK2 (p44 and p42) are known to play a pivotal role in signaling events in the brain where they are involved in neuronal plasticity (11), neuronal survival (12), and processing the amyloid precursor protein (13,14). They play a crucial role in learning and memory, including long term potentiation (15,16) and long term spatial memory (17).…”