Organophosphorus compound effects on neurotrophin receptors and intracellular signaling

Neurite outgrowth of SH-SY5Y neuroblastoma cells following the addition of spinal cord extracts from chickens exposed to a neuropathic organophosphorus (OP) compound suggests the presence of a growth factor during OP neuropathy. However, exposure of SH-SY5Y cells directly to neuropathic OP compounds results in apoptosis and/or decreased neurite outgrowth. These cellular effects may follow OP-induced interference with neurotrophin-receptor binding and/or intracellular signaling resulting from receptor binding. We hypothesized that sub-lethal concentrations of a neuropathic OP compound interferes with neurotrophin-receptor binding as well as specific intracellular signaling pathways in neuroblastoma cells which would not occur with a non-neuropathic OP compound. SH-SY5Y cells were exposed to a neuropathic OP compound (PSP; 0.01, 0.1, 1.0 μM), a neuropathic OP compound with nerve growth factor (1.0 μM PSP + 1 ng/ml NGF), a non-neuropathic OP compound (paraoxon; 100 μM), and medium only for 4, 8, 24, and 48 h. Western blots indicate that cells exposed to a low dose of PSP or the high dose of PSP + NGF contained the phosphorylated form of a common neurotrophin receptor (pp75) that was four times greater than that of the phosphorylated form of the high-affinity NGF receptor (pTrkA) suggesting that p75 activation may contribute to early cell death after exposure to OP compounds. Furthermore, events in signaling pathways after exposure to PSP differed from those after exposure to paraoxon, with activation of the MEK1/2 protein increasing significantly only after exposure to paraoxon. Both types of OP compounds, however, caused significant activation of Akt in the PI-3K cell-survival pathway. These results suggest that exposure to a non-neuropathic OP compound causes increased activity of the MAPK pathway whereas exposure to neuropathic OP compounds prevented upregulation of the pathway. Since this pathway is integral to neurite outgrowth and cell survival, this study has revealed molecular mechanisms implicated in neuronal response after exposure to neuropathic OP compounds.