EFFECT OF CHONDROITINASE ABC ON AXONAL MYELINATION AND GLIAL SCAR AFTER SPINAL CORD INJURY IN RATS_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

ZHANG Tao , SHEN Yixin , LU Leilei , FAN Zhihai , HUO Weiwei.

Department of Orthopaedics, the Second Affiliated Hospital of Soochow University, Suzhou Jiangsu, 215004, P.R.China. Corresponding author: SHEN Yixin, E-mail: shenyx@mail.szldbz.gov.cn;

Corresponding author：

Keywords：

Spinal cord injury; Chondroitinase ABC; Myelin basic protein; Growth associated protein 43; Glial fibrillary acidic protein; Rat

Video：

Export PDF Favorites Scan Get Citation

Abstract Full text Figures/Tables Video References Cited by

Objective To investigate the effects of chondroitinase ABC (ChABC) on axonal myelination and glial scar after spinal cord injury (SCI) in rats. Methods Seventy-two adult male Sprague Dawley rats were randomly assigned into ChABC treatment group (group A), saline treatment group (group B), and sham operation group (group C), 24 rats in each group. In groups A and B, the SCI model was established with modified Allen’s method and then the rats of groups A and B were administrated by subarachnoid injection of 6 μL ChABC (1 U/mL) and saline respectively at 1 hour after injury and every day for 1 week; the rats of group C served as control, which canal was opened without damage to spinal cord. At 1, 7, 14, and 28 days after operation, the locomotor functions were evaluated according to the Basso-Beattie-Bresnahan (BBB) score scale; and the spinal cord samples were harvested for HE staining, Nissl staining, and immunohistochemistry analysis to detect the change of myelin basic protein (MBP), growth associated protein 43 (GAP-43), and glial fibrillary acidic protein (GFAP) of the injured spinal cord. Results At different time points, the BBB score of group C was significantly higher than those of groups A and B (P lt; 0.05), and the BBB score of group A was significantly better than that of group B at 14 and 28 days after operation (P lt; 0.05). HE staining and Nissl staining showed that the morphous and the neuron number of the remainant injured spinal cord in group A were better than those in group B. The integral absorbance (IA) values of MBP and GAP-43 and the positive area of GFAP after SCI in groups A and B were significantly higher than those in group C at different time points (P lt; 0.05), and the IA values of MBP and GAP-43 were significantly higher in group A than those in group B at 7, 14, and 28 days after operation (P lt; 0.05), but the positive area of GFAP was significantly smaller in group A than that in group B (P lt; 0.05). Conclusion The ChABC can effectively improve the microenvironment of the injured spinal cord of rats, enhance the expressions of MBP and GAP-43, and inhibit the expression of GFAP, which promotes the axonal regeneration and myelination, attenuate glial scar formation, and promote the recovery of nerve function.

Citation： ZHANG Tao,SHEN Yixin,LU Leilei,FAN Zhihai,HUO Weiwei.. EFFECT OF CHONDROITINASE ABC ON AXONAL MYELINATION AND GLIAL SCAR AFTER SPINAL CORD INJURY IN RATS. Chinese Journal of Reparative and Reconstructive Surgery, 2013, 27(2): 145-150. doi: Copy

1.	Cafferty WB, Yang SH, Duffy PJ, et al. Functional axonal regeneration through astrocytic scar genetically modified to digest chondroitin sulfate proteoglycans. J Neurosci, 2007, 27(9): 2176-2185.
2.	Cakir E, Usul H, Peksoylu B, et al. Effect of citicoline on experimental spinal cord injury. J Clin Neurosci, 2005, 12(8): 923-926.
3.	Yaksh TL, Kohl RL, Rudy TA. Induction of tolerance and withdrawal in rats receiving morp- hine in the spinal subarachnoid space. Eur J Pharmacol, 1977, 42(3): 275-284.
4.	Basso DM, Beattie MS, Bresnahan JC. A sensitive and reliable locomotor rating scale for open field testing in rats. J Neurotrauma, 1995, 12(1): 1-21.
5.	Siebert JR, Osterhout DJ. The inhibitory effects of chondroitin sulfate proteog- lycans on oligodendrocytes. J Neurochem, 2011, 119(1): 176-188.
6.	Monnier PP, Sierra A, Schwab JM, et al. The Rho/ROCK pathway mediates neurite growth- inhibitory activity associated with the chondroitin sulfate proteoglycans of the CNS glial scar. Mol Cell Neurosci, 2003, 22(3): 319-330.
7.	Ughrin YM, Chen ZJ, Levine JM. Multiple regions of the NG2 proteoglycan inhibit neurite growth and induce growth cone collapse. J Neurosci, 2003, 23(1): 175-186.
8.	Caggiano AO, Zimber MP, Ganguly A, et al. Chondroitinase ABC improves locomotion and bladder function following contusion injury of the rat spinal cord. J Neurotrauma, 2005, 22(2): 226-239.
9.	Tester NJ, Howland DR. Chondroitinase ABC improves basic and skilled locomotion in spinal cord injured cats. Exp Neurol, 2008, 209(2): 483-496.
10.	Lee H, McKeon RJ, Bellamkonda RV. Sustained delivery of thermostabilized chABC enhances axonal sprouting and functional recovery after spinal cord injury. Proc Natl Acad Sci U S A, 2010, 107(8): 3340-3345.
11.	Bradbury EJ, Carter LM. Manipulating the glial scar: Chondroitinase ABC as a therapy for spinal cord injury. Brain Res Bull, 2011, 84(4-5): 306-316.
12.	Harauz G, Ishiyama N, Hill CM, et al. Myelin basic protein-diverse conformational states of an intrinsically unstructured protein and its roles in myelin assembly and multiple sclerosis. Micron, 2004, 35(7): 503-542.
13.	吴波, 孙磊, 任先军. 少突胶质前体细胞移植治疗对大鼠脊髓损伤轴突髓鞘化的影响. 中华创伤杂志, 2010, 26(11): 1035-1039.
14.	Atkinson S, Li YQ, Wong CS. Changes in oligodendrocytes and myelin gene expression after radiation in the rodent spinal cord. Int J Radiat Oncol Biol Phys, 2003, 57(4): 1093-1100．.
15.	Fukuda A, Fukuda H, Swanpalmer J, et al. Age-dependent sensitivity of the developing brain to irradiation is correlated with the number and Vulnerability of progenitor cells. J Neurochem, 2005, 92(3): 569-584.
16.	Kotter MR, Li WW, Zhao C, et al. Myelin impairs CNS remyelination by inhibiting oligoden- drocyte precursor cell differentiation. J Neurosci, 2006, 26(1): 328-332.
17.	徐纪伟, 孙丹华, 刘喜民, 等. 硫酸软骨素酶ABC对大鼠脊髓损伤后生长相关蛋白43和神经胶质纤维酸性蛋白的影响. 中国修复重建外科杂志, 2010, 24(10): 1212-1216.
18.	Metz GA, Schwab ME. Behavioral characterization in a comprehensive mouse test battery reveals motor and sensory impairments in growth-associated protein-43 null mutant mice. Neuroscience, 2004, 129(3): 563-574.
19.	Shen Y, Mani S, Donovan SL, et al. Growth-associated protein-43 is required for commissural axon guidance in the developing vertebrate nervous system. J Neurosci, 2002, 22(1): 239-247.
20.	Horner PJ, Gage FH. Regenerating the damaged central nervous system. Nature, 2000, 407(6807): 963-970.
21.	Rhodes KE, Moon LD, Fawcett JW. Inhibiting cell proliferation during formation of the glial scar: effect on axon regeneration in the CNS. Neuroscience, 2003, 120(1): 41-56.
22.	Tang X, Davies JE, Davies SJ. Changes in distribution, cell associations, and protein expression levels of NG2, neurocan, phosphacan, brevican, verslcan V2, and tenascin-C during acute to chronic maturation of spinal cord scar tissue . J Neurosci Res, 2003, 71 (3): 427-444.
23.	Wang XX, Hasan O, Arzeno A, et al. Axonal regeneration induced by blockade of glial inhibitors coupled with activation of intrinsic neuronal growth pathways. Exp Neurol, 2012, 237 (1): 55-69.
24.	Starkey ML, Bartus K, Barritt AW, et al. Chondroitinase ABC promotes compensatory sprouting of the intact corticospinal tract and recovery of forelimb function following unilateral pyramidotomy in adult mice. Eur J Neurosci, 2012. [Epub ahead of print].
25.	Bosch KD, Bradbury EJ, Verhaagen J, et al. Chondroitinase ABC promotes plasticity of spinal reflexes following peripheral nerve injury. Exp Neurol, 2012, 238 (1): 64-78.

1. Cafferty WB, Yang SH, Duffy PJ, et al. Functional axonal regeneration through astrocytic scar genetically modified to digest chondroitin sulfate proteoglycans. J Neurosci, 2007, 27(9): 2176-2185.
2. Cakir E, Usul H, Peksoylu B, et al. Effect of citicoline on experimental spinal cord injury. J Clin Neurosci, 2005, 12(8): 923-926.
3. Yaksh TL, Kohl RL, Rudy TA. Induction of tolerance and withdrawal in rats receiving morp- hine in the spinal subarachnoid space. Eur J Pharmacol, 1977, 42(3): 275-284.
4. Basso DM, Beattie MS, Bresnahan JC. A sensitive and reliable locomotor rating scale for open field testing in rats. J Neurotrauma, 1995, 12(1): 1-21.
5. Siebert JR, Osterhout DJ. The inhibitory effects of chondroitin sulfate proteog- lycans on oligodendrocytes. J Neurochem, 2011, 119(1): 176-188.
6. Monnier PP, Sierra A, Schwab JM, et al. The Rho/ROCK pathway mediates neurite growth- inhibitory activity associated with the chondroitin sulfate proteoglycans of the CNS glial scar. Mol Cell Neurosci, 2003, 22(3): 319-330.
7. Ughrin YM, Chen ZJ, Levine JM. Multiple regions of the NG2 proteoglycan inhibit neurite growth and induce growth cone collapse. J Neurosci, 2003, 23(1): 175-186.
8. Caggiano AO, Zimber MP, Ganguly A, et al. Chondroitinase ABC improves locomotion and bladder function following contusion injury of the rat spinal cord. J Neurotrauma, 2005, 22(2): 226-239.
9. Tester NJ, Howland DR. Chondroitinase ABC improves basic and skilled locomotion in spinal cord injured cats. Exp Neurol, 2008, 209(2): 483-496.
10. Lee H, McKeon RJ, Bellamkonda RV. Sustained delivery of thermostabilized chABC enhances axonal sprouting and functional recovery after spinal cord injury. Proc Natl Acad Sci U S A, 2010, 107(8): 3340-3345.
11. Bradbury EJ, Carter LM. Manipulating the glial scar: Chondroitinase ABC as a therapy for spinal cord injury. Brain Res Bull, 2011, 84(4-5): 306-316.
12. Harauz G, Ishiyama N, Hill CM, et al. Myelin basic protein-diverse conformational states of an intrinsically unstructured protein and its roles in myelin assembly and multiple sclerosis. Micron, 2004, 35(7): 503-542.
13. 吴波, 孙磊, 任先军. 少突胶质前体细胞移植治疗对大鼠脊髓损伤轴突髓鞘化的影响. 中华创伤杂志, 2010, 26(11): 1035-1039.
14. Atkinson S, Li YQ, Wong CS. Changes in oligodendrocytes and myelin gene expression after radiation in the rodent spinal cord. Int J Radiat Oncol Biol Phys, 2003, 57(4): 1093-1100．.
15. Fukuda A, Fukuda H, Swanpalmer J, et al. Age-dependent sensitivity of the developing brain to irradiation is correlated with the number and Vulnerability of progenitor cells. J Neurochem, 2005, 92(3): 569-584.
16. Kotter MR, Li WW, Zhao C, et al. Myelin impairs CNS remyelination by inhibiting oligoden- drocyte precursor cell differentiation. J Neurosci, 2006, 26(1): 328-332.
17. 徐纪伟, 孙丹华, 刘喜民, 等. 硫酸软骨素酶ABC对大鼠脊髓损伤后生长相关蛋白43和神经胶质纤维酸性蛋白的影响. 中国修复重建外科杂志, 2010, 24(10): 1212-1216.
18. Metz GA, Schwab ME. Behavioral characterization in a comprehensive mouse test battery reveals motor and sensory impairments in growth-associated protein-43 null mutant mice. Neuroscience, 2004, 129(3): 563-574.
19. Shen Y, Mani S, Donovan SL, et al. Growth-associated protein-43 is required for commissural axon guidance in the developing vertebrate nervous system. J Neurosci, 2002, 22(1): 239-247.
20. Horner PJ, Gage FH. Regenerating the damaged central nervous system. Nature, 2000, 407(6807): 963-970.
21. Rhodes KE, Moon LD, Fawcett JW. Inhibiting cell proliferation during formation of the glial scar: effect on axon regeneration in the CNS. Neuroscience, 2003, 120(1): 41-56.
22. Tang X, Davies JE, Davies SJ. Changes in distribution, cell associations, and protein expression levels of NG2, neurocan, phosphacan, brevican, verslcan V2, and tenascin-C during acute to chronic maturation of spinal cord scar tissue . J Neurosci Res, 2003, 71 (3): 427-444.
23. Wang XX, Hasan O, Arzeno A, et al. Axonal regeneration induced by blockade of glial inhibitors coupled with activation of intrinsic neuronal growth pathways. Exp Neurol, 2012, 237 (1): 55-69.
24. Starkey ML, Bartus K, Barritt AW, et al. Chondroitinase ABC promotes compensatory sprouting of the intact corticospinal tract and recovery of forelimb function following unilateral pyramidotomy in adult mice. Eur J Neurosci, 2012. [Epub ahead of print].
25. Bosch KD, Bradbury EJ, Verhaagen J, et al. Chondroitinase ABC promotes plasticity of spinal reflexes following peripheral nerve injury. Exp Neurol, 2012, 238 (1): 64-78.

Chinese Journal of Reparative and Reconstructive Surgery

EFFECT OF CHONDROITINASE ABC ON AXONAL MYELINATION AND GLIAL SCAR AFTER SPINAL CORD INJURY IN RATS

Abstract Full text Figures/Tables Video References Cited by

Previous Article

Next Article

Format

Content