Staurosporin induces neurite outgrowth through ROS generation in HN33 hippocampal cell lines

Abstract

Staurosporin, a specific inhibitor of PKC, is widely used in studies of signal transduction pathways. Previous studies have shown that staurosporin induces neurite outgrowth, but the underlying mechanisms remain unclear. Here we report that staurosporin induces neurite outgrowth in HN33 hippocampal cells. Two other PKC inhibitors, Go 6976 (specific for alpha- and beta-isoforms) and rotterlin (a selective inhibitor of PKC delta), have no neuritogenic effect. In addition, staurosporin specifically increases ROS generation. NAC, which inhibits the generation of ROS, suppresses the staurosporin-induced neurite outgrowth in HN33 cells. Further, H(2)O(2) causes neurite outgrowth. Taken together, these results confirm a neuritogenic effect of staurosporin and point to ROS as the signal mediator of staurosporin-induced neurite outgrowth in HN33 hippocampal cells. Theme: Development and regeneration Topic: Neurotrophic factors: receptors and cellular mechanisms.

Critical role of the JNK‐p53‐GADD45α apoptotic cascade in mediating oxidative cytotoxicity in hippocampal neurons

“…When cells were pretreated with 1 mmol·L -1 N-acetyl-cysteine (NAC), a scavenger of ROS, for 1 h before treatment with glutamate for additional 8 h, the accumulation of intracellular ROS was suppressed ( Figure 4C, left and right), confirming earlier observations (Min et al, 2006;Fukui et al, 2009). As shown in Figure 4D, immunofluorescence staining showed that NAC treatment inhibited glutamate-induced p53 activation and reduced the levels of GADD45a mRNA ( Figure 4E, left panel) and protein ( Figure 4E, right panel), suggesting that ROS generation is upstream of the activation of p53 and GADD45a.…”

Hydrogen Peroxide Promotes Injury-Induced Peripheral Sensory Axon Regeneration in the Zebrafish Skin

“…In zebrafish, H 2 O 2 may be signaling directly to axons, altering the extracellular matrix, or eliciting a second signal from keratinocytes to promote axon growth, but does not require leukocytes (Figure 8). Assessing whether application of H 2 O 2 to somatosensory neurons in culture can improve axon growth, as has been reported for hippocampal neurons in culture [33], could help resolve whether H 2 O 2 acts directly or indirectly on axons to influence their regeneration.…”

“…It will be interesting to determine whether H 2 O 2 also plays a role in these phenomena. Intriguingly, H 2 O 2 improves hippocampal neurite outgrowth in culture [33]. In C. elegans , a mutation in pxn-2 , which encodes an extracellular peroxidase, improves regeneration of mechanosensory axons [34].…”

Cell Models for the Study of Sex Steroid Hormone Neurobiology

“…Two commonly used hippocampal-derived cell lines are the HT22 and the HN33 cells [107,108]. Our laboratory found mRNA for ERα, ERβ, PGRMC1, and GPR30 and no mRNA PR, and AR in HT-22 cells.…”