Rotenone induced neurodegeneration is mediated via cytoskeleton degradation and necroptosis

Rotenone has widespread beneficial effects in agriculture, fisheries and animal husbandries; however prolonged exposure causes a detrimental effect on the health of personnel working in such industries. Rotenone during its extraction, formulation or usage may cross the blood brain barrier leading to neurodegeneration and the development of Parkinson's disease like symptoms. It is a known inhibitor of the mitochondrial ETC complex I and responsible for impairing the OXPHOS system. Our study showed that rotenone exposure results in an increased production of ROS and decreased ATP level along with a conspicuous loss of mitochondrial membrane potential in N2A cells. The transcription and expression pattern of cofilin, a key component of actin cytoskeleton, was also altered after rotenone exposure; leading to the actin cytoskeleton degradation. We further observed an increased expression, as well as activity of matrix metalloproteinase9 (MMP9) in rotenone exposed N2A cells; suggesting the involvement of inflammation upon rotenone exposure. Simultaneously, an opposite pattern was noticed for the tissue inhibitors of metalloproteinases-1 (TIMP-1) protein, which is a known modulator of MMP9 activity. Additionally, the localization of MMP9 along with alpha-synuclein, UCHL1 and cofilin suggested their close proximity and cross interaction upon rotenone treatment. Furthermore, we observed significant increase in the level of TNF-alpha upon rotenone exposure along with the phosphorylation of RIPK1, RIPK3 and MLKL that has been identified as the necroptosis markers leading to programmed necroptotic death.

Automated summary

Rotenone has both beneficial effects in agriculture, fisheries and animal husbandries, as well as detrimental effects on the health of personnel in these industries. Prolonged exposure to rotenone can lead to neurodegeneration and the development of Parkinson's disease symptoms. Our study showed that rotenone exposure in N2A cells results in increased ROS production, decreased ATP levels, and loss of mitochondrial membrane potential. The expression of cofilin, a key component of actin cytoskeleton, was altered and the actin cytoskeleton degraded. MMP9 expression and activity increased, indicating inflammation upon rotenone exposure. TNF-alpha levels and necroptosis markers also increased, suggesting programmed necroptotic death.