Involvement of autophagy in tri-ortho-cresyl phosphate- induced delayed neuropathy in hens.

Autophagy is a highly conserved cellular self-degradative process that plays a housekeeping role in removing aggregated proteins and damaged organelles. Our recent work has found that tri-ortho-cresyl phosphate (TOCP), a neuropathic organophosphate (OP), decreased the level of beclin 1 (a key molecule in the process of autophagy) in hen nerve tissues (Song et al., 2012). However, the role of autophagy in the pathogenesis of organophosphorus ester-induced delayed neuropathy (OPIDN) remains unclear. Here, we investigated whether dysfunctional autophagy was associated with the initiation and development of TOCP-induced delayed neuropathy. Adult hens were given a single dose of 750mg/kg TOCP (p.o.) and sacrificed on days 1, 5, 10, and 21 after dosing, respectively. The formation of autophagosomes in spinal cord motor neurons was observed by transmission electron microscopy, the level of autophagy-related proteins in hen spinal cords and tibial nerves was determined by Western blot analysis. The results demonstrated that the number of autophagosomes was markedly increased in the myelinated and unmyelinated axons of hen spinal cords after TOCP exposure. In the meantime, the level of two molecular markers for autophagy, microtubule-associated protein light chain-3 (LC3) and p62/SQSTM1 in hen nerve tissues was significantly decreased and increased, respectively. Furthermore, a marked reduction in autophagy-regulated proteins including ULK 1, AMBRA 1, ATG 5, ATG 7, ATG 12 and VPS34 expression was also observed. Our results suggested that the administration of TOCP resulted in a significant inhibition of autophagy activity in neurons, which might be associated with the pathogenesis of OPIDN.