臺灣博碩士論文加值系統

周邊神經末梢受傷後，受傷神經的遠端殘肢(distal stump of injured nerve)部份會進行髓鞘及軸突內細胞骨架崩解的病理變化現象，此現象稱之為瓦勒氏退化(Wallerian degeneration) 。一般的囓齒類動物，此病理變化發生時程非常快速；此實驗中所使用的實驗動物具有緩慢退化特質(slow degeneration)的突變種老鼠─C57BL/Ola mice (Ola mice)。Ola mice突變的位置在第四對體染色體(autosome)遠端，稱之為wlds的對偶基因(allele)上。實驗中，我們分別用兩種不同的神經損傷模式─截斷(transection)及夾傷(crush)來引發老鼠的坐骨神經(sciatic nerve)進行瓦勒氏退化。手術後不同時間對老鼠的足墊 (footpad) 及足底肌肉 (plantar muscles) 以免疫細胞化學染色(immunocytochemistry)觀察以比較不同生理功能神經進行瓦勒氏退化與神經再生的時程。發現當老鼠的坐骨神經受到損傷後，進行瓦勒氏退化的速度在不同生理功能的神經並不相同；運動神經最快(神經受損後兩週)，感覺神經次之(神經受損後三週)，自主神經為最慢(神經受損後五週)。同時，遠端殘肢的末端部位(terminal end of distal stump of injured nerve)進行瓦勒氏退化的速度會比近端(proximal portion of distal stump of injured nerve)來的快。此外，因為Ola mice神經退化速度緩慢，所以也進而間接影響受傷神經的再生速度，使得神經再生的時程比一般的囓齒類動物所需時間上更長。綜合以上觀察，推知不同類型神經進行瓦勒氏退化與神經再生所可能包含的機制並不完全相同。

Wallerian degeneration is a common pathological phenomenon after peripheral nerve injury. Wallerian degeneration is an extremely slow process in the C57BL/Ola (Ola) mice, a substrain of C57BL, carrying a mutant allele, Wlds in chromosome 4. In this study, we transected and crushed sciatic nerves of Ola mice and compared Wallerian degeneration among motor, sensory and autonomic nerves. The speeds of degeneration were different among different types of nerves. Degeneration of motor nerve terminals inside neuromuscular junctions were the fastest, with all nerves disappearing within two weeks. By three weeks, all the epidermal nerves were degenerated. Sweat glands innervation disappeared within five weeks after nerve injury. The terminal portion of injured sciatic nerves were degenerated faster than the proximal portion of the injured nerves. The speeds of regeneration in Ola mice were slower than those in wild-type mice. These results raise the possibility that mechanisms of Wallerian degeneration could be different among different types of nerves.

Ⅰ 英文摘要........................................................1.
Ⅱ 中文摘要.........................................................2.
Ⅲ 引言.....................................................3.
Ⅳ 材料與方法..........................................6.
Ⅴ 結果.....................................................9.
Ⅵ 總結...................................................17.
Ⅶ 討論...................................................21.
Ⅷ 附錄...................................................27.
(i)圖表與說明...................................28.
(ii)參考資料......................................42.

Brown, M. C., Perry, V. H., Lunn, E. R., Gordon, S. & Heumann, R. (1991) Macrophage dependence of peripheral sensory nerve regeneration: possible involvement of nerve growth factor. Neuron 6: 359-370.
Brown, M. C., Lunn, E. R. & Perry, V. H. (1992) Consequences of slow Wallerian degeneration for regeneration motor and sensory axons. Journal of Neurobiology 23: 521-536.
Brown, M. C., Perry, V. H., Hunt, S. P. & Lapper, S. R. (1993) Further studies on motor and sensory nerve regeneration in mice with delayed Wallerian degeneration. European Journal of Neuroscience 6: 420-428.
Dailey, A. T., Avellino, A. M., Benthem, L., Silver, J. & Kliot, M. (1998) Complement depletion reduced macrophage infiltration and activation during Wallerian degeneration and axonal regeneration. The Journal of Neuroscience 18: 6713-6722.
Dalsgaard, C. J., Rydh, M. & Hagerstrand, A. (1989) Cutaneous innervation in man visualized with protein gene product 9.5 (PGP 9.5) antibodies. Histochemistry 92: 385-389.
Fruttiger, M., Schachner, M. & Martini, R. (1995) Tenascin-C expression during Wallerian degeneration in C57BL/Wlds mice: possible implications for axonal regeneration. Journal of neurocytology 24: 1-14.
Geffen, L. B. & Hughes, C. C. (1972) Degeneration of sympathetic nerves in vitro and development of smooth muscle supersensitivity to noradrenaline. Journal of Physiology 221: 71-84.
George, E. B., Glass, J. D. & Griffin, J. W. (1995) Axotomy-induced axonal degeneration is mediated by calcium influx through ion-specfic channels. The journal of Neuroscience 15: 6445-6452.
Glass, J. D., Brushart, T. M., George, E. B. & Griffin, J. W. (1993) Prolonged survival of transected nerve fibers in C57BL/Ola mice is an intrinsic characteristic of the axon. Journal of Neurocytology 22: 311-321.
Glass, J. D., Schryer, B. L. & Griffin, J. W. (1994) Calcium-mediated degeneration of the axonal cytoskeleton in the Ola mice. Journal of Neurochemistry 62: 2472-2475.
Glass, J. D.,N, N., Dry, I., Culver, A. I. & Wesselingh, S. (1998) Cloning of m-calpain 80 KD subunit from the axonal degeneration-resistant WLD(S) mouse mutant. Journal of Neuroscience Research 52: 653-660.
Griffin, J. W., George, B. D., Hsieh, S. T. & Glass, J. D. (1995) Axonal degeneration and disorders of the axonal cytoskeleton. THE AXON: Structure, Function and Pathology. (edited by Waxman, S. G., Kocsis, J. D. & Stys, P. K.) pp.375-390. New York Oxford: Oxford university press.
Hall, S. M. (1993) Observation on the progress of Wallerian degeneration in transected peripheral nerves of C57BL/Wld mice in the presence of recruited macrophages. Journal of Neurocytology 22: 480-490.
Hilliges, M., Wang, L. & Johansson, O. (1995) Ultrastructural evidence for nerve fibers within all vital layers of the human epidermis. Journal of Investigative Dermatology 104: 134-137.
Hsieh, S. T., Choi, S., Lin, W. M., Chang, Y. C., Mcarthur, J. C. & Griffin, J. W. (1996) Epidermal denervation and its effects on keratinocytes and Langerhans cells. Journal of Neurocytology 25: 512-523.
Karanth, S. S., Springall, D. R., Kuhn, D. M., Levene, M. M. & Polak, J. M. (1991) An immunocytochemical study of cutaneous innervation and the distribution of neuropeptides and protein gene product 9.5 in man and commonly employed laboratory animals. The American Journal of Anatomy 191: 369-383.
Kamakura, K., Ishiura, S., Sugita, H. & Toyokura, Y. (1983) Identification of Ca2+-activated neutral protease in the peripheral nerve and its effects on neurofilament degeneration. Journal of Neyrochemistry 40: 908-913.
Kennedy, W. R. & Wendelschafer-Crabb, G. (1993) The innervation of human epidermis. Journal of Nurological Science 115: 184-190.
Lin, W. M., Hsieh, S. T., Huang, I. T., Griffin, J. W. & Chen, W. P. (1997) Ultrastructural localization and regulation of protein gene product 9.5. NeuroReport 8: 2999-3004.
Lubinska, L. (1982) Pattern of Wallerian degeneration of myelinated fibres in short and long peripheral stumps and in isolated segments of rat and phrenic nerve. Interpretation of the role of axoplasmic flow of the trophic factor. Brain Research 233: 227-240.
Lunn, E. R., Perry, V. H., Brown, M. C., Rosen, H. & Gordon, S. (1989) Absence of Wallerian degeneration does not hinder regeneration in peripheral nerve. European Journal of Neuroscience 1: 27-33.
Lunn, E. R., Brown, M. C. & Perry, V. H. (1990) The pattern of axonal degeneration in the peripheral nervous system varies with different types of lesion. Neuroscience 35: 157-165.
Lyon, M. F., Ogunkolade, B. W., Brown, M. C., Atherton, D. J. & Perry, V. H. (1993) A gene affecting Wallerian nerve degeneration maps distally on mouse chromosome 4. Proc. Natl. Acad. Sci. USA 90: 9717-9720.
McCarthy, B. G., Hsieh, S. T., Stocks, A. (1995) Cutaneous innervation in sensory neuropathies: Evaluation by skin biopsy. Neurology 45: 1848-1855.
Navarro, X., Verdu, E., Wendelschafer-Crabb, G. & Kennedy, W. R. (1995) Innervation of cutaneous structures in the mouse hind paw: A confocal microscopy immunohistochemical study. Journal of Neuroscience Research 41: 111-120.
Okamoto, M. & Riker, W. F. Jr. (1969) Motor nerve terminals as the site of initial functional changes after denervation. Journal of General Physiology 53: 70-80.
Ribcherter, R. R., Tsao, J. W., Barry, J. A., Asgari-Jirhandeh, N., Perry, V. H. & Brown, M. C. (1995) Persistence of neuromuscular junctions after axotomy in mice with slow Wallerian degeneration (C57BL/Wlds).
European Journal of Neuroscience 7: 1641-1650.
Schlaepfer, W. W. (1974) Calcium-induced degeneration of axoplasma in isolated segments of rat phrenic nerve. Brain Research 69: 203-215.
Stankovic, N., Johansson, O. & Hildebrand, C. (1996) Occurrence of epidermal nerve ending in glabrous and hairy skin of the rat foot after sciatic nerve regeneration. Cell and Tissue Research 284: 161-166.
Thompson, R. J., Doran, J. F., Jackson, P., Dhillon, A. P. & Rode, J. (1983) PGP 9.5-a new marker for vertebrate neurons and neuroendocrine cells. Brain Research 278: 224-228.
Tsao, J. W., Brown, M. C., Carden, M. J., McLean, W. G. & Perry, V. H. (1993) Loss of the compound action potential: an electrophysiological, biochemical and Morphological study of early events in axonal degeneration in the C57BL/Ola mice. European Journal of Neuroscience 6: 516-524.
Wang, L., Hilliges, M., Jernmerg, T., Wiegleb-Edstrom, D. & Johansson, O. (1990) Protein gene product 9.5-immunoreactive nerve fibers and cells in human skin. Cell and Tissue Research 261: 25-33.
Wilkinson, K. D., Lee, K. & Desphand, S. (1989) The neuron-specific protein PGP 9,5 is a ubiqutin carboxyl-terminal hydrolase. Science 246: 670-673.
Zimmerman, U.-J. P. & Schlaepfer, W. W. (1982) Characterization of a brain calcium-activated protease that degrades neurofilament proteins. Biochemistry 21: 3977-3983.