Decelerated transport and its mechanism of 2,5-hexanedione on middle-molecular-weight neurofilament in rat dorsal root ganglia cells.
Chronic exposure to n-hexane induces peripheral-central axonopathy, mediated by its metabolite 2,5-hexanedione (2,5-HD), in occupational workers and experimental animals, but the underlying mechanism is still unclear. In the current study, we investigated the effects of 2,5-HD on middle-molecular-weight neurofilament (NF-M) axonal transport using live-cell imaging technique in cultured rat dorsal root ganglia (DRG) cells. PA-GFP-NF-M plasmid was transfected into DRG neurons and live-cell imaging was performed to observe the slow axonal transport of NF-M. The levels of cytoskeleton and motor proteins in DRG cells were detected by Western-blot and the concentration of ATP was determined using an ATP Assay Kit. The results showed that 2,5-HD administration resulted in a decrease of NF-M axonal transport and a reduction of three neurofilament subunits levels in DRG cells. Furthermore, 2,5-HD exposure significantly decreased ATP contents and the protein levels of kinesin heavy chain (KHC). These findings indicated that 2,5-HD reduced slow axonal transport, neurofilaments cargoes, motor proteins and ATP energy in rat DRG cells, which may contribute to 2,5-HD-induced neurotoxicity.
Institute of Toxicology, Shandong University, Jinan 250012, China; College of Life Sciences, Shandong Normal University, Jinan 250014, China.
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Han
Department of Toxicology, Shandong Center for Disease Control and Prevention, Jinan 250014, China.
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Cheng
Institute of Toxicology, Shandong University, Jinan 250012, China.
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Song
Institute of Toxicology, Shandong University, Jinan 250012, China.
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Zeng
Institute of Toxicology, Shandong University, Jinan 250012, China.
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An
Institute of Toxicology, Shandong University, Jinan 250012, China. Electronic address: keqinx@sdu.edu.cn.
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D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001370:Axonal Transport / Q000187:drug effects* / Q000502:physiology; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D003598:Cytoskeletal Proteins / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005727:Ganglia, Spinal / Q000187:drug effects* / Q000502:physiology; D006588:Hexanones / Q000494:pharmacology*; D015151:Immunoblotting; D020409:Molecular Motor Proteins / Q000378:metabolism; D016900:Neurofilament Proteins / Q000378:metabolism*; D009498:Neurotoxins / Q000494:pharmacology*; D017207:Rats, Sprague-Dawley
D003598:Cytoskeletal Proteins; D006588:Hexanones; D020409:Molecular Motor Proteins; D016900:Neurofilament Proteins; D009498:Neurotoxins; C061195:neurofilament protein M; D000255:Adenosine Triphosphate; C011269:2,5-hexanedione
2,5-hexanedione; motor proteins; neurofilaments; slow axonal transport