Axonopathy-inducing 1,2-diacetylbenzene forms adducts with motor and cytoskeletal proteins required for axonal transport.
The aromatic hydrocarbon 1,2-diacetylbenzene (1,2-DAB) is a protein-reactive gamma-diketone metabolite of the neurotoxic solvent 1,2-diethylbenzene (1,2-DEB). The effect of neurotoxic 1,2-DAB and its non-neurotoxic isomer 1,3-DAB has been studied on motor proteins and cytoskeletal proteins of rat spinal cord (SC). For in vitro studies, SC slices were incubated with 1, 2, 5, 10 mM of DAB isomers for 30 min at 37 degrees C. For in vivo studies, rats received (i.p.) 20 mg/kg/day of 1,2-DAB or 1,3-DAB, or vehicle (2% acetone in saline), 5 days a week for 2 weeks. Spinal cord and sciatic nerve proteins were subjected to Western blotting using monoclonal mouse antibodies to NF-M, kinesin, dynein, and tau. Proteins were quantified and paired mean comparisons performed to assess concentration-dependent changes in native protein bands. In vitro, 1,2-DAB produced a concentration-dependent decrease of motor and cytoskeletal proteins. While dynein and tau appeared similarly affected by 1,2-DAB, kinesin was most affected by the toxicant. In vivo, 1,2-DAB affected motor and cytoskeletal proteins of sciatic nerves and spinal cord differentially. In general, sciatic nerve proteins were much more affected than spinal cord proteins. The results show that motor proteins that drive axonal transport anterogradely (kinesin) and retrogradely (dynein), cytoskeletal protein NF-M, which is slowly transported in the anterograde direction, and microtubule-associated protein, tau, which is involved in axonal transport, are differentially impacted by 1,2-DAB. By contrast, non-neurotoxic isomer 1,3-diacetylbenzene (1,3-DAB), had no adverse effect on neural proteins either in vitro or in vivo. 2D-Differential in gel electrophoresis (2D-DIGE) of sciatic nerves from neurotoxic 1,2-DAB and non-neurotoxic 1,3-DAB treated rats revealed 197 and 304 protein spots, respectively.
Sabri
Mohammad I
MI
Center for Research on Occupational & Environmental Toxicology, L606, Oregon Health & Science University, Portland, OR 97239, USA. sabri@ohsu.edu
Hashemi
Seyed B
SB
Lasarev
Michael R
MR
Spencer
Peter S
PS
D000098:Acetophenones / Q000633:toxicity*; D000818:Animals; D001370:Axonal Transport / Q000187:drug effects / Q000502:physiology*; D001369:Axons / Q000187:drug effects / Q000473:pathology*; D015153:Blotting, Western; D003598:Cytoskeletal Proteins / Q000378:metabolism* / Q000502:physiology; D004398:Dyneins / Q000378:metabolism; D016547:Kinesins / Q000378:metabolism; D008297:Male; D009046:Motor Neurons / Q000187:drug effects / Q000378:metabolism*; D009419:Nerve Tissue Proteins / Q000378:metabolism* / Q000502:physiology; D020258:Neurotoxicity Syndromes / Q000378:metabolism* / Q000473:pathology; D051381:Rats; D017207:Rats, Sprague-Dawley; D012584:Sciatic Nerve / Q000378:metabolism; D013116:Spinal Cord / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D016875:tau Proteins / Q000378:metabolism
D000098:Acetophenones; D003598:Cytoskeletal Proteins; D009419:Nerve Tissue Proteins; D016875:tau Proteins; C048792:1,2-diacetylbenzene; D004398:Dyneins; D016547:Kinesins