Degenerative Changes of the Rat Dorsal Root Ganglion (DRG) Cells Following a Tight Spinal Nerve Ligation

랫드 척수신경 결찰에 따른 척수신경절세포의 퇴행성변화

  • Kim, Yi-Suk (Department of Anatomy, Ewha Womans University School of Medicine) ;
  • Jo, Seung-Mook (Department of Anatomy, Gachon Medicine and Science University)
  • 김이석 (이화여자대학교 의학전문대학원 해부학교실) ;
  • 조승묵 (가천의과학대학교 기초의학부 해부학과)
  • Received : 2009.08.25
  • Accepted : 2009.09.22
  • Published : 2009.09.30

Abstract

This study aim to disclose a possible mechanism for the neuronal cell death induced by peripheral nerve injury following a spinal nerve ligation (SNL) as a neuropathic pain model. Male Sprague-Dawley rats (270~290 g) were used for this study. Pain threshold was evaluated for their response to mechanical (von Frey hairs) stimuli 1, 3, and 7 days after a tight ligation of L5 ventral ramus. In control group, the small ganglion cells were strongly stained with routine toluidine blue (TB), whereas the large ganglion cells showed a little bit weak stainity. Each large ganglion cell is surrounded by perineuronal satellite cells. In experimental groups, small ganglion cells showing apparent degenerative changes increased on 1 day, and showed a peak in degenerative cell number at 3 days group, and decreased gradually at 7 days group. We also found a small number of large-sized ganglion cells showing mild degenerative changes. However their satellite cells ware relatively intact with no typical findings throughout this experiment. Under the electron microscope, small ganglion cells showed various stage and typical features of the dark degeneration including mitochondrial swelling.

본 연구는 잘 알려진 통증모델을 대상으로 척수신경절세포의 변성과정을 경시적으로 관찰하고자 하였다. 10주령된 Sprague-Dawley 계통의 랫드를 실험동물로 사용하였고, pentobarbital (50mg/kg)로 마취상태에서 다섯째 허리신경(L5)의 앞가지를 결찰한 후 1일, 3일, 7일 실험군으로 구분하였다. 실험 1일과 3일군에서 작은 신경절세포에서 퇴행성변화가 먼저 관찰되었고, 중간 및 큰 신경절세포에서는 그 변화가 미약했다. 7일 실험군에서는 작은 신경절세포의 수가 크게 감소하였던 반면, 중간 및 큰 신경절세포에서는 큰 변화가 없었다. 전자현미경 소견으로는 작은 신경절세포의 경우 초기 1일 및 3일군에서 미토콘드리아의 능선의 변형과 부종이 관찰되었으며, 세포질은 검게 변성되었다. 반면 큰 신경절세포의 경우 모든 실험군에서 변형된 세포가 관찰되지 않았으며, 소기관들도 잘 보존되었다. 이상의 결과를 요약하면, 척수신경 결찰에 의한 통증자극으로 랫드 척수신경절 내 작은 신경절세포는 다른 신경절세포에 비해 퇴행성변화가 빠르게 나타났으며, 미토콘드리아 등 세포질소기관의 부종을 동반한 암적화변성(dark degeneration)을 통해 사멸되었다.

Keywords

References

  1. Besson JM: The neurobiology of pain. Lancet 353 : 1610-1615, 1999 https://doi.org/10.1016/S0140-6736(99)01313-6
  2. Chard PS, Bleakman D, Savidge JR, Miller RJ: Capsaicin-induced neurotoxicity in cultured dorsal root ganglion neurons: involvement of calcium-activated proteases. Neuroscience 65 : 1099-1108, 1995 https://doi.org/10.1016/0306-4522(94)00548-J
  3. Dykes RW: Parallel processing of somatosensory information: a theory. Brain Res Rev 6 : 47-116, 1983 https://doi.org/10.1016/0165-0173(83)90004-8
  4. Furst S: Transmitters involved in antinociception in the spinal cord. Brain Res Bull 48 : 129-141, 1999 https://doi.org/10.1016/S0361-9230(98)00159-2
  5. Goff JR, Burkey AR, Goff DJ, Jasmin L: Reorganization of the spinal dorsal horn in modela of chronic pain: correlation with behaviour. Neurosci 82 : 559-574, 1998 https://doi.org/10.1016/S0306-4522(97)00298-4
  6. Gould HJ 3rd, England JD, Soignier RD, Nolan P, Minor LD, Liu ZP, Levinson SR, Paul D: Ibuprofen blocks changes in Na v 1.7 and 1.8 sodium channels associated with complete Freund's adjuvant-induced inflammation in rat. J Pain 5 : 270-280, 2004 https://doi.org/10.1016/j.jpain.2004.04.005
  7. Henken DB, Goldstein ME, Martin JR: Herpes simplex virus type-2 infection by a footpad route results in neuronal death in mouse spinal ganglia. J Neurol Sci 115 : 177-183, 1993 https://doi.org/10.1016/0022-510X(93)90222-K
  8. Inglis JJ, Nissim A, Lees DM, Hunt SP, Chernajovsky Y, Kidd BL:The differential contribution of tumour necrosis factor to thermal and mechanical hyperalgesia during chronic inflammation. Arthritis Res Ther 7 : R807-816, 2005 https://doi.org/10.1186/ar1743
  9. Ji RR, Zhang X, Wiesenfeld-Hallin Z, Hokfelt T: Expression of neuropeptide Y and neuropeptide Y (Y1) receptor mRNA in rat spinal cord and dorsal root ganglia following peripheral tissue inflammation. J Neurosci 14 : 6423-6434, 1994
  10. Jo SM, Danscher G, Schroder HD, Suh SW: Depletion of vesicular zinc in dorsal horn of spinal cord causes increased neuropathic pain in mice, Biometals (in press), 2007 https://doi.org/10.1007/s10534-007-9103-x
  11. Jo SM, Nam SA, Yoon SS, Ko BM, Choi CD, Choi WB, Chun M:Indoleamine-accumulating cell death and endogenous glial cell reaction induced by 5,7-dihydroxytryptamine in the the cat retina. J Korean Med Sci 13 : 414-422, 1998 https://doi.org/10.3346/jkms.1998.13.4.414
  12. Kato N, Nemoto K, Arino H, Fujikawa K: Influence of peripheral inflammation on growth-associated phosphoprotein (GAP-43) expression in dorsal root ganglia and on nerve recovery after crush injury. Neurosci Res 45 : 297-303, 2003 https://doi.org/10.1016/S0168-0102(02)00234-1
  13. Kim SJ, Choi YH, Yu YC, Lee B, Jo SM: Decrease in zinc concentration in the rat spinal gray matter induced by peripheral nerve ligation, and its relation with increased pain threshold. Korean J Anat 37(4) : 369-375, 2004. (Korean)
  14. Klein CM, Guillanmondegui O, krenek CD, La Forte RA, Coggeshall RE: Do neuropeptides in the dorsal horn changes if the dorsal root ganglion cell death that normally accompanies peripheral nerve transaction is prevented. Brain Res 552(2) : 273-282, 1991 https://doi.org/10.1016/0006-8993(91)90092-A
  15. Lee B, Jun G, Kim YS, Lee B, Lee YI, Kim DJ, Jo SM: Alteration of ionic zinc distribution in rat spinal ganglion by inflammatory pain stimuli: Autometallography. Korean J Anat 38(6) : 561-566, 2005
  16. Lee SE, Shen H, Taglialatela G, Chung JM, Chung KS : Expression of nerve growth factor in the dorsal root ganglion after peripheral nerve injury. Brain Res 796 : 99-106, 1998 https://doi.org/10.1016/S0006-8993(98)00335-7
  17. Lekan HA, Chung K, Yoon YW, Chung JM, Coggeshall RE: Loss of dorsal root ganglion cells concomitant with dorsal horn root axon sprouting following segmental nerve lesions. Neurosci 81 : 527-534, 1997 https://doi.org/10.1016/S0306-4522(97)00173-5
  18. Mapp PI, Terenghi G, Walsh DA, Chen ST, Cruwys, Garrett N, Kidd BL, Polak JM, Blake DR: Monoarthritis in the rat knee induces bilateral and time-dependent changes in substance p and calcitonin gene-related peptide immunoreactivity in the spinal cord. Neurosci 57 : 1091-1096, 1993 https://doi.org/10.1016/0306-4522(93)90051-G
  19. Papassotiropoulos A, Ludwig M, Naib-Majani W, Rao GS: Induction of apoptosis and secondary necrosis in rat dorsal root ganglion cell cultures by oxidized low density lipoprotein. Neurosci Lett 3 : 33-36, 1996 https://doi.org/10.1016/0304-3940(76)90095-1
  20. P$\acute{e}$rez-Castejón MC, Vera-Gil A, Lahoz M, Aisa J, Recreo MP, Pes N, Serrano P, Barral MJ: Location of Zinc and 65Zn in spinal ganglia of the rat. Histol Histopathol 17 : 799-903, 2002
  21. Pierucci A, de Oliveira AL: Increased sensory neuron apoptotic death 2 weeks after peripheral axotomy in C57BL/6J mice compared to A/J mice. Neurosci Lett 27 : 127-131, 2006
  22. Ren K, Dubner R: NMDA receptor antagonists attenuate mechanical hyperalgesia in rat with unilateral inflammation of the hindpaw. Neurosci Lett 163 : 22-26, 1993 https://doi.org/10.1016/0304-3940(93)90220-F
  23. Sluka KA, Westlund KN: Behavioral and immunohistochemical changes in an experimental arthritis model in rats. Pain 55 : 367-377, 1993 https://doi.org/10.1016/0304-3959(93)90013-F
  24. Szoke E, Seress L, Szolcsanyi J: Neonatal capsaicin treatment result in prolonged mitochondrial damage and delated cell death of B cells in the rat trigeminal ganglia. Neuroscience 113 : 925-937, 2002 https://doi.org/10.1016/S0306-4522(02)00208-7
  25. Takeda M, Tanimoto T, Nasu M, Ikeda M, Kadoi J, Matsumoto S: Activation of NK1 receptor of trigeminal root ganglion via substance P paracrine mechanism contributes to the mechanical allodynia in the temporomandibular joint inflammation in rats. Pain 116 : 375-85, 2005 https://doi.org/10.1016/j.pain.2005.05.007
  26. Velazquez RA, Cai YJ, Shi Q, Larson AA: The distribution of zinc selenite and expression of metallothionein III mRNA in the spinal cord and dorsal root ganglia of the rat suggest a role for zinc in sensory transmission. J Neurosci 19 : 2288-2300, 1999
  27. Wang H, Zhang RX, Wang R, Qiao JT: Decreased expression of Nmethyl-D-aspartate (NMDA) receptors in rat dorsal root ganglion following complete Freund's adjuvant-induced inflammation:an immunocytochemical study for NMDA NR1 subunit. Neurosci Lett 23 : 195-198, 1999
  28. Zimmermann M: Pathobiology of neuropathic pain. Eur J Pharmacol 19 : 23-37, 2001