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Time-dependent Alteration of Cytoskeletal Proteins in Cerebral Cortex of Rat During 2,5-Hexanedione-induced Neuropathy

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Abstract

To investigate the mechanisms of the axonopathy induced by 2,5-hexanedione (2,5-HD), male Wistar rats were administered at a dosage of 400 mg/kg/day 2,5-HD (five times per week). The rats produced a slightly, moderately, or severely abnormal neurological changes, respectively, after 2, 4, or 8 weeks of treatment. The cerebrums were Triton-extracted and ultracentrifuged to yield a pellet fraction and a corresponding supernatant fraction. The relative levels of six cytoskeletal proteins (NF-L, NF-M, NF-H, α-tubulin, β-tubulin, and β-actin) in both fractions were determined by immunoblotting. The results showed that NFs content in HD-treated rats demonstrated a progressive decline as the intoxication of HD continued. As for microtubule proteins, the levels of α-tubulin and β-tubulin demonstrated some inconsistent changes. The content of α-tubulin kept unchangeable, while the content of β-tubulin increased significantly at the late stage of HD exposure. Furthermore, the content of β-actin in both fractions remained unaffected throughout the study. These findings suggest that HD intoxication resulted in a progressive decline of NFs, which was highly correlated with the development of HD-induced neuropathy.

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References

  1. Altenkirch H, Mager J, Stoltenburg G, Helmbrecht J (1977) Toxic polyneuropathies after sniffing a glue thinner. J Neurol 214:137–152

    Article  PubMed  CAS  Google Scholar 

  2. Spencer PS, Schaumburg HH, Sabri MI, Veronesi B (1980) The enlarging view of hexacarbon neurotoxicity. Crit Rev Toxicol 7:279–356

    PubMed  CAS  Google Scholar 

  3. Cavanagh JB, Bennetts RJ (1981) On the pattern of changes in the rat nervous system produced by 2,5 hexanediol. A topographical study by light microscopy. Brain 104:297–318

    Article  PubMed  CAS  Google Scholar 

  4. Couri D, Milks M (1982) Toxicity and metabolism of the neurotoxic hexacarbons n-hexane, 2-hexanone, and 2,5-hexanedione. Annu Rev Pharmacol Toxicol 22:145–166

    Article  PubMed  CAS  Google Scholar 

  5. Lehning EJ, Dyer KS, Jortner BS, LoPachin RM (1995) Axonal atrophy is a specific component of 2,5-hexanedione peripheral neuropathy. Toxicol Appl Pharmacol 135:58–66

    Article  PubMed  CAS  Google Scholar 

  6. Lehning EJ, Jortner BS, Fox JH, Arezzo JC, KitanoT, LoPachin RM (2000) Gamma-diketone peripheral neuropathy. I. Quality morphometric analyses of axonal atrophy and swelling. Toxicol Appl Pharmacol 165:127–140

    Article  PubMed  CAS  Google Scholar 

  7. Opanashuk LA, He DK, Lehning EJ, Lopachin RM (2001) Diketone peripheral neuropathy III. Neurofilament gene expression. Neurotoxicology 22:215–220

    Article  PubMed  CAS  Google Scholar 

  8. DeCaprio AP, O’Neill EA (1985) Alterations in rat axonal cytoskeletal proteins induced by in vitro and in vivo 2,5-hexanedione exposure. Toxicol Appl Pharmacol 78:235–247

    Article  PubMed  CAS  Google Scholar 

  9. Carden MJ, Lee VM, Schlaepfer WW (1986) 2,5-Hexanedione neuropathy is associated with the covalent crosslinking of neurofilament proteins. Neurochem Pathol 5:25–35

    PubMed  CAS  Google Scholar 

  10. Lapadula DM, Suwita E, Abou-Donia MB (1988) Evidence for multiple mechanisms responsible for 2,5-hexanedione-induced neuropathy. Brain Res 458:123–131

    Article  PubMed  CAS  Google Scholar 

  11. LoPachin RM, He D, Reid ML, Opanashuk LA (2004) 2,5-Hexanedione-induced changes in monomeric neurofilament protein content of rat spinal cord fractions. Toxicol Appl Pharmacol 198:61–73

    Article  PubMed  CAS  Google Scholar 

  12. LoPachin RM, He D, Reid ML (2005) 2,5-Hexanedione-induced changes in the neurofilament subunit pools of rat peripheral nerve. Neurotoxicology 26:229–240

    Article  PubMed  CAS  Google Scholar 

  13. Zhang T, Han X, Zhao X, Zhao L, Zhang C, Yu L, Yu S, Xie K (2005) 2,5-Hexanedione induced reduction in protein content and mRNA expression of neurofilament in rat cerebral cortex. Environ Toxicol Pharmacol 20:92–98

    Article  CAS  Google Scholar 

  14. LoPachin RM, Ross JF, Reid ML, Das S, Mansukhani S, Lehning EJ (2002) Neurological evaluation of toxic axonopathies in rats: acrylamide and 2,5-hexanedione. Neurotoxicology 23:95–110

    Article  PubMed  CAS  Google Scholar 

  15. Chiu FC, Norton WT (1982) Bulk preparation of CNS cytoskeleton and the separation of individual neurofilament proteins by gel filtration: dye-binding characteristics and amino acid compositions. J Neurochem 39:1252–1260

    Article  PubMed  CAS  Google Scholar 

  16. Tsuda M, Tashiro T, Komiya Y (1997) Increased solubility of highmolecular-mass neurofilament subunit by suppression of dephosphorylation: its relation to axonal transport. J Neurochem 68:2558–2565

    Article  PubMed  CAS  Google Scholar 

  17. Lee MK, Cleveland DW (1996) Neuronal intermediate filaments. Annu Rev Neurosci 19:187–217

    Article  PubMed  CAS  Google Scholar 

  18. Lee MK, Xu Z, Wong PC, Cleveland DW (1993) Neurofilaments are obligate heteropolymers in vivo. J Cell Biol 122:1337–1350

    Article  PubMed  CAS  Google Scholar 

  19. Cleveland DW, Monteiro MJ, Wong PC, Gill SR, Gearhart JD, Hoffman PN (1991) Involvement of neurofilaments in the radial growth of axons. J Cell Sci Suppl 15:85–95

    PubMed  CAS  Google Scholar 

  20. Lee MK, Cleveland DW (1994) Neurofilament function and dysfunction: involvement in axonal growth and neuronal disease. Curr Opin Cell Biol 6:34–40

    Article  PubMed  CAS  Google Scholar 

  21. Micheva KD, Vallee A, Beaulieu C, Herman IM, Leclerc N (1998) Beta-actin is confined to structures having high capacity of remodeling in developing and adult rat cerebellum. Eur J Neurosci 10:3785–3798

    Article  PubMed  CAS  Google Scholar 

  22. Shea TB, Dahl DC, Nixon RA, Fischer I (1997) Triton-soluble phosphovariants of the heavy neurofilament subunit in developing and mature mouse central nervous system. J Neurosci Res 48:515–523

    Article  PubMed  CAS  Google Scholar 

  23. Shea TB, Sihag RK, Nixon RA (1990) Dynamics of phosphorylation and assembly of the high molecular weight neurofilament subunit in NB2a/d1 neuroblastoma. J Neurochem 55:1784–1792

    Article  PubMed  CAS  Google Scholar 

  24. Cohlberg JA, Hajarian H, Tran T, Alipourjeddi P, Noveen A (1995) Neurofilament protein heterotetramers as assembly intermediates. J Biol Chem 270:9334–9339

    Article  PubMed  CAS  Google Scholar 

  25. Arbuthnott ER, Boyd IA, Kalu KU (1980) Ultrastructural dimensions of myelinated peripheral nerve fibers in the cat and their relation to conduction velocity. J Physiol 308:125–137

    PubMed  CAS  Google Scholar 

  26. Sakaguchi T, Okada M, Kitamura T, Kawasaki K (1993) Reduced diameter and conduction velocity of myelinated fibers in the sciatic nerve of a neurofilament-deficient mutant quail. Neurosci Lett 153:65–68

    Article  PubMed  CAS  Google Scholar 

  27. Banik NL, Hogan EL, Powers JM, Whetstine LJ (1982) Degradation of cytoskeletal proteins in spinal cord injury. Neurochem Res 7:1465–1475

    Article  PubMed  CAS  Google Scholar 

  28. Banik NL, Matzelle DC, Gantt-Wilford G, Osborne A, Hogan EL (1997) Increased calpain content and progressive degradation of neurofilament protein in spinal cord injury. Brain Res 752:301–306

    Article  PubMed  CAS  Google Scholar 

  29. Ray SK, Hogan EL, Banik NL (2003) Calpain in the pathophysiology of spinal cord injury: neuroprotection with calpain inhibitors. Brain Res Rev 42:169–185

    Article  PubMed  CAS  Google Scholar 

  30. Guo-Ross S, Yang E, Bondy SC (1998) Elevation of cerebral proteases after systemic administration of aluminum. Neurochem Int 33(3):277–282

    Article  PubMed  CAS  Google Scholar 

  31. Gupta RP, Abou-Donia MB (1996) Alterations in the neutral proteinase activities of central and peripheral nervous systems of acrylamide-, carbon disulfide-, or 2,5-hexanedione-treated rats. Mol Chem Neuropathol 29:53–66

    Article  PubMed  CAS  Google Scholar 

  32. Rao MV, Houseweart MK, Williamson TL, Crawford TO, Folmer J, Cleveland DW (1998) Neurofilament-dependent radial growth of motor axons and axonal organization of neurofilaments does not require the neurofilament heavy subunit (NF-H) or its phosphorylation. J Cell Biol 143:171–181

    Article  PubMed  CAS  Google Scholar 

  33. Moskowitz PF, Smith R, Pickett J, Frankfurter A, Oblinger MM (1993) Expression of the class III β-tubulin gene during axonal regeneration of rat dorsal root ganglion neurons. J Neurosci Res 34:129–134

    Article  PubMed  CAS  Google Scholar 

  34. Song FY, Yu SF, Zhao XL, Zhang CL, Xie KQ (2006) Carbon disulfide-induced changes in cytoskeleton protein content of rat cerebral cortex. Neurochem Res 31:71–79

    Article  PubMed  CAS  Google Scholar 

  35. Yagihashi S, Kamijo M, Watanabe K (1990) Reduced myelinated fiber size correlates with loss of axonal neurofilaments in peripheral nerve of chronically streptozotocin rats. Am J Pathol 136:1365–1373

    PubMed  CAS  Google Scholar 

  36. Liuzzi FJ, Bufton SM, Vinik AI (1998) Streptozotocin-induced diabetes mellitus causes changes in primary sensory neuronal cytoskeletal mRNA levels that mimic those caused by axotomy. Exp Neurol 154:381–388

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

This work was supported by grants from the Ministry of Science and Technology of China (No.2002CB512907), and National Natural Science Fund of China (No. 271138).

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Authors and Affiliations

  1. Institute of Toxicology, Shandong University, 44 West Wenhua Road, Jinan, Shandong, 250012, People’s Republic of China

    Fuyong Song, Cuili Zhang, Sufang Yu, Xiulan Zhao, Lihua Yu & Keqin Xie

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  1. Fuyong Song
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  2. Cuili Zhang
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  3. Sufang Yu
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  4. Xiulan Zhao
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  5. Lihua Yu
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Correspondence to Keqin Xie.

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Song, F., Zhang, C., Yu, S. et al. Time-dependent Alteration of Cytoskeletal Proteins in Cerebral Cortex of Rat During 2,5-Hexanedione-induced Neuropathy. Neurochem Res 32, 1407–1414 (2007). https://doi.org/10.1007/s11064-007-9325-x

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  • DOI: https://doi.org/10.1007/s11064-007-9325-x

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