An elevated level of cyclic AMP (cAMP) within cells activates gene expression through the cAMP-PKA-CREB pathway. Among the CREB target genes, some immediate early genes exist that are responsive to cAMP including the nur77 and c-fos genes. Treatment with dibutyryl-cAMP (dbcAMP) as well as nerve growth factor (NGF) induces neurite outgrowth in PC12 cells. Here, we report that acetylation of histone H3 was gradually stimulated after treatment with dbcAMP in PC12 cells and peaked 1 h after treatment. As the result of reverse transcription-polymerase chain reaction (RT-PCR) and quantitative real-time PCR (qPCR) experiments, both nur77 and c-fos gene expression were found to have peak 1 h after treatment. Knock-down with siRNA against nur77 mRNA inhibited the neurite outgrowth induced by dbcAMP, whereas knock-down with siRNA against c-fos mRNA did not inhibit the dbcAMP-induced neurite outgrowth. A chromatin immunoprecipitation (ChIP) assay revealed that the nur77 gene was associated with the acetylated Lys14 of histone H3 after treatment with dbcAMP. However, the amount of c-fos gene associated with acetylated histone H3 was not changed after treatment with dbcAMP. These results suggest that the expression of nur77, which is essential for the neuronal differentiation induced by dbcAMP, is up-regulated via dbcAMP-induced acetylation of the Lys14 of histone H3 in PC12 cells.

No abstract

No abstract

We previously reported that nicotine protected against tunicamycin (Tm)-induced ER stress-mediated apoptosis, but not thapsigargin (Tg)-induced apoptosis in PC12 cells. In the present study, we report that the expression of glucose-regulated protein 78 (GRP78) was suppressed by nicotine in Tm-treated PC12 cells. Interestingly, the GRP78 expression was not changed by nicotine in Tg-treated cells. Moreover, nicotine reduced the activation of caspase-12 in Tm-treated cells, but not in Tg-treated cells. These results suggest that nicotine prevented Tm-induced ER stress-mediated apoptosis by attenuating an early stage of Tm-induced ER stress. It was possible that the suppression of GRP78 expression by nicotine was achieved through the suppression of the Ire1-XBP1 and/or ATF6 pathways. We observed that nicotine suppressed the Tm-induced, but not Tg-induced, splicing of XBP1 mRNA, and also suppressed the Tm-induced, but not Tg-induced, production of cleaved ATF6 in PC12 cells. These results indicate that the suppression of Ire1-XBP1 and ATF6 pathways contributes to the suppression of GRP78 expression by nicotine in Tm-treated PC12 cells, suggesting that nicotine suppresses a common step upstream of both the Ire1-XBP1 and ATF6 pathways which are required for the expression of GRP78 during Tm-induced ER stress.

Recent reports have indicated that some low-molecular-weight compounds mimic neurotrophic factors inducing neurite outgrowth and neuroprotection. Carnosic acid (CA) promotes neurite outgrowth through the activation of Nrf2 in PC12 cells. CA also protects neurons via the keap/Nrf2 transcriptional pathway from oxidative stress. Forskolin-induced neurite outgrowth is mediated by activation of the PKA signalling pathway and this PKA-mediated neurite outgrowth is achieved by the expression of nur77 in PC12 cells. In addition, forskolin at its low concentration is closely related to the cAMP-induced protective function against L-DOPA-induced cytotoxicity in PC12 cells. A HDAC inhibitor trichostatin A (TSA) increases neurite length via p53 acetylation in rat cultured cerebellar granule neurons and in cerebral cortical neurons, and also protects neurons against glutathione depletion-induced oxidative stress. Recently, it was revealed that Nrf2 and p53 bind to CBP/p300 directly, and Nur77 is acetylated in vivo and in vitro by CBP/p300. Acetylation of Nrf2, p53 and Nur77 by CBP/p300 may constitute a novel similar regulatory mechanism for low-molecular-weight compounds with neurotrophic activities.