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Induction of neurite-outgrowth in PC12 cells by alpha-toxin from Clostridium perfringens
Masataka Oda1, Yuki Saito, Yuka Morimune
1Department of Microbiology, Faculty of Pharmaceutical Science, Tokushima Bunri University, Yamashiro-cho, Tokushima 770-8514, Japan. masa@ph.bunri-u.ac.jp
Abstract:
Alpha-toxin-induced phosphorylation of PDK1 via the tyrosine kinase A (TrkA) receptor signaling pathway plays an important role in the activation of rabbit neutrophils. The relation between the toxin and TrkA, however, remains poorly understood. Here, we show that the toxin-induced phosphorylation of TrkA is closely related to the induction of neurite-outgrowth in PC12 cells. The toxin induced neurite-outgrowth and phosphorylation of TrkA in the cells in a dose-dependent manner. K252a, a TrkA inhibitor, and shRNA for TrkA inhibited the toxin-induced neurite-outgrowth, and phosphorylation of TrkA and ERK1/2. PD98059, an inhibitor of the ERK1/2 cascade, inhibited phosphorylation of ERK1/2 and the neurite-outgrowth induced by alpha-toxin. The wild-type toxin induced the formation of diacylglycerol, and neurite-outgrowth, but H148G, a variant toxin which binds to cell membranes and has lost the enzymatic activity did not. We demonstrated that the phosphorylation of TrkA through the phospholipid metabolism induced by the toxin synergistically play a key role in neurite-outgrowth.
Insights
Alpha-toxin triggers nerve cell growth by activating the tyrosine kinase A (TrkA) receptor pathway, involving ERK1/2 signaling and phospholipid metabolism for neurite-outgrowth.
Area of Science:
- Neuroscience
- Cell Biology
- Biochemistry
Background:
- Alpha-toxin activates rabbit neutrophils via tyrosine kinase A (TrkA) receptor signaling.
- The precise mechanism linking alpha-toxin to TrkA activation and subsequent cellular responses is not fully understood.
Purpose of the Study:
- To investigate the role of alpha-toxin in TrkA receptor phosphorylation and neurite-outgrowth in PC12 cells.
- To elucidate the signaling pathways involved in alpha-toxin-induced neuronal differentiation.
Main Methods:
- PC12 cells were treated with varying concentrations of alpha-toxin.
- Inhibition studies were performed using K252a (TrkA inhibitor) and PD98059 (ERK1/2 inhibitor).
- TrkA and ERK1/2 phosphorylation, neurite-outgrowth, and diacylglycerol formation were assessed.
Main Results:
- Alpha-toxin dose-dependently induced TrkA phosphorylation and neurite-outgrowth in PC12 cells.
- TrkA inhibition (K252a, shRNA) blocked toxin-induced neurite-outgrowth and ERK1/2 phosphorylation.
- ERK1/2 inhibition (PD98059) reduced neurite-outgrowth and ERK1/2 phosphorylation.
- Wild-type alpha-toxin induced diacylglycerol formation and neurite-outgrowth, unlike a non-enzymatic variant.
Conclusions:
- Alpha-toxin-induced TrkA phosphorylation, mediated by phospholipid metabolism, is crucial for neurite-outgrowth in PC12 cells.
- The ERK1/2 cascade is a key downstream effector in the alpha-toxin-TrkA signaling pathway leading to neuronal differentiation.
