【摘 要】
迷走感覺神經元對呼吸道及其他臟器的神經性調控，扮演重要的角色。但其感覺神經元在迷走神經節的區域分布並不確定。本研究將HRP注射進入右側胸迷走神經及塗敷在右側喉返神經斷端，使之逆向運送貯於迷走神經節細胞，再利用diaminobenzidine (DAB)反應呈現標示的神經節細胞。此外，以神經毒素(RCA-60)注射進入右側胸迷走神經中，使之逆向運送至迷走神經節細胞並破壞核醣體，再利用capsaicin誘發呼吸道神經性血漿外洩，並計算被India ink標示的血管面積表示其發炎程度。
目前的研究發現，在迷走神經節中提供感覺神經纖維於胸迷走神經的神經元比例，除了節狀神經節近咽神經處外，並沒有明顯的區域性分布，節狀及頸神經節的分布分別為42.2 % 及30.5 %。而支配喉返神經的神經元比例，在兩個神經節中分別為13.3 % 及8.3 %。以RCA-60注射一及二週後，發現節狀神經節的神經元呈顯著的變性變化，細胞變蒼白，Nissl’s bodies消失，細胞質出現空泡化變性，並且對於同側支氣管樹的神經性血漿外洩有明顯的抑制作用，血管面積密度減少了71至89 %；此表示RCA-60選擇性的破壞分佈於同側支氣管樹的迷走感覺神經元。
節狀及頸迷走神經節提供了不同數量的感覺神經纖維至其分支，以調控內臟器官的生理機能。

【Abstract】
Vagal sensory neurons play an important role in the neural control of airway and other visceral organs. Regional distribution of vagal sensory neurons in the vagal ganglia that correspond to different viscera is uncertain. Horseradish peroxidase (HRP) was applied to the right thoracic vagus nerve and recurrent laryngeal nerve to be retrogradely transported to the neurons in vagal nodose and jugular ganglia. Labeling of neuronal cell bodies was visualized with diaminobenzidine reaction. Ricinus communis agglutinin-60 (RCA-60) was injected into the right thoracic vagus nerve and transported retrogradely to cause destruction of the ribosome in the vagal ganglia neuron cell bodies. The magnitude of neurogenic plasma extravasation induced by capsaicin was measured by the area density of the India ink-labeled leaky blood vessels in the trachea and bronchi.
The present study demonstrated that there was no distinct localization of HRP- labeled neurons, except at the level of pharyngeal nerve that was connected to the middle of the nodose ganglion. There were approximately 42.2 % of labeled neurons in the nodose ganglion and 30.5 % in the jugular ganglion 3 days after application of HRP in the thoracic vagus nerve. After application of HRP to the recurrent laryngeal nerve, 13.3 % of labeled neurons was found in the nodose ganglion and 8.3 % in the jugular ganglion. One to two weeks after RCA injection into the thoracic vagus nerve, many cell bodies of neurons had a striking degenerative alteration and the cytoplasmic density was markedly reduced. Nissl’s bodies obviously disappeared and vacuoles were the usual feature. Application of RCA-60 also inhibited the neurogenic plasma extravasation in the right bronchial tree that were reduced by 71- 89 %. It is suggested that RCA- 60 selectively destroyed the vagal sensory neurons that innervated the ipsilateral branchial tree.
It is concluded that vagal nodose and jugular ganglia supplied different number of sensory neurons to the vagal branches and regulated the physiological function of the visceral organs.

【目 錄】
誌謝-----------------------------------------------------------------------------------------i
中文摘要-----------------------------------------------------------------------------------ii
英文摘要----------------------------------------------------------------------------------iii
目錄----------------------------------------------------------------------------------------iv
圖目錄--------------------------------------------------------------------------------------v
文獻探討-----------------------------------------------------------------------------------1
研究目的-----------------------------------------------------------------------------------9
材料與方法
I. 實驗動物-------------------------------------------------------------------------------10
II. 藥物配製------------------------------------------------------------------------------10
III. 以藥物局部處理迷走神經分支--------------------------------------------------12
IV. 辣椒素誘發呼吸道之神經性發炎反應-----------------------------------------14
V. 組織處理與切片製作---------------------------------------------------------------16
VI. 過氧化

【參 考 文 獻】
Atasever, A., Palaoglu, S., Erbengi, A. & Celik, H.H. (1994) Labelling of neurons in the rat superior cervical ganglion after injection of wheat- germ agglutinin-horseradish peroxidase into the contralateral ganglion: evidence of transneuronal labelling. Journal of Anatomy. 184: 93-7
Barbieri, L., Battelli, M.G. & Stirpe, F. (1993) Ribosome-inactivating proteins from plants. Biochimica et Biophysica Acta. 1154: 237-282
Bergren, D.R. (1988) Capsaicin challenge, reflex broncho-constriction, and local action of substance-P. American Journal of Physiology. 254: 845-852
Bisby, M.A. (1982) Function of retrograde axonal transport. Federation Proceedings. 41: 2307-2311
Bousquet, J., Chanez, P., Lacoste, J.Y., White, R., Vic, P., Godard, P. & Michel, F.B. (1992) Asthma : a disease remodeling the airways. Allergy. 47: 3-11
Brokaw, J.J., Hillenbrand, C.M.,White, G.W. & McDonald D.M. (1990) Mechanism of tachyphylaxis associated with neurogenic plasma extravasation in the rat trachea. American Review of Respiratory Disease. 141: 1434-1440
Brown, R.F. & White, D.E. (1997) Ultrastructure of rat lung following inhalation of ricin aerosol. International Journal of Experimental Pathology. 78: 267-276
Carobi, C. (1996) A quantitative investigation of the effects of neonatal capsaicin treatment on vagal afferent neurons in the rat. Cell and Tissue Research. 283: 305-311
Cevolani, D., Strocchi, P., Bentivoglio, M. & Stirpe, F. (1995) Suicide retrograde transport of volkensin in cerebellar afferents: direct evidence, neuronal lesions and comparison with ricin. Brain Research. 689: 163-171
Christian, E.P., Togo, J.A., Naper, K.E., Koschorke, G., Taylor, G. A. & Weinreich, D. (1993) A retrograde labeling technique for the functional study of airway-specific visceral afferent neurons. Journal of Neuroscience Methods. 47: 147-160
De Castro, F., Sanchez-Vives, M.V., Munoz-Martinez, E.J. & Gallego, R. (1995) Effect of postganglionic nerve section an synaptic transmission in the superior cervical ganglion of the guinea pig. Neuroscience. 67: 689-695
Dray, A., Patel, I.A., Perkins, M.N. & Rueff, A. (1992) Bradykinin-induced activation of nociceptors: receptor and mechanistic studies on the neonatal rat spinal cord-tail preparation in vitro. British Journal of Pharmacology. 107: 1129-1134
Fitzgerald, M. (1983) Capsaicin and sensory neurons- a review. Pain. 15:109-130
Furusawa, K., Yasuda, K., Okuda, D., Tanaka, M. & Yamaoka, M. (1996) Central distribution and peripheral functional properties of afferent and efferent components of the superior laryngeal nerve: morphological and electrophysiological studies in the rat. The Journal of Comparative Neurology. 375: 147-156
Grant, G. (1993) Projection patterns of primary sensory neurons studied by transganalionic methods: somatotopy and target-related organization. Brain Research Bulletin. 30: 199-208
Hanamori, T. & Smith, D.V. (1986) Central projections of the hamster superior laryngeal nerve. Brain Research Bulletin. 16: 271-279
Heath, P.R., Chessell, I.P., Sanders, M.W., Francis, P.T., Bowen, D.M. & Pearson, R.C. (1997) Neuronal degeneration by suicide transport following injection of volkensin into rat cerebral cortex. Experimental Neurology. 147: 192-203
Hisa, Y., Toyoda, K., Uno, T., Murakami, Y. & Ibata, Y. (1991) Localization of the sensory neurons in the canine nodose ganglion sending fibers into the internal branch of the superior laryngeal nerve. European Archives of Otorhinolaryngology 248: 265-267
Hiura, A. & Ishizuka, H. (1989) Changes in features of degenerating primary sensory neurons with time after capsaicin treatment. Acta Neuropathologica. 78: 35-46
Hogg, J.C., James, A.L. & Pare, P.D. (1991) Evidence for inflammation in asthma. American Review of Respiratory Disease. 143: 39-42
Hsieh, J.H., Chen, R.F., Wu, J.J., Yen, C.T. & Chai, C.Y. (1998) Vagal innervation of the gastrointestinal tract arises from dorsal motor nucleus whiel that of the heart largely from nucleus ambiguus in cat. Journal of the Autonomic Nervous System. 70: 38-50
Huang, H.T. (1993) Unilateral cervical vagotomy decreases the magnitude of neurogenic inflammation induced by capsaicin in the ipsilateral bronchial tree of rats. Anatomy and Embryology. 188: 363-370
Huang, H.T., Huang, S.H. & Lour, Y.G. (1995) Postvagotomy changes in neurogenic plasma extravasation in rat bronchi. Journal of the Autonomic Nervous System. 55: 9-17
Hunter, D.D. & Undem, B.J. (1999) Identification and substance P content of vagal afferent neurons innervating the epithelium of the guinea pig trachea. American Journal of Respiratory and Critical Care Medicine. 159: 1943-1948
Hwang, T., Huang, H.T. & Tsao, C.F. (1999) Thoracic vagus section distal to the recurrent laryngeal nerve reduces substance P-immunoreactive innervation in the rat bronchial tree. Anatomy and Embryology. 200: 153-160
Kalia, M. & Mesulam, M.M. (1980) Brain stem projections of sensory and motor components of the vagus complex in the cat: II. Laryngeal, tracheobronchial, pulmonary, cardiac and gastrointestinal branches. The Journal of Comparative Neurology. 193: 467-508
Kressel, M. (1998) Tyramide amplification allows anterograde tracing by horseradish peroxidase- conjugated lectins in conjuction with simultaneous immunohistochemistry. Journal of Histochemistry and Cytochemistry. 46: 527-533
Kressel, M. & Radespiel-Troger, M.R. (1999) Anterograde tracing and Immunohistochemical characterization of potentially mechanosensitive vagal afferents in the esophagus. The Journal of Comparative Neurology. 412: 161-172
Kuo, H.P., Rhode, J.A.L., Tokuyama, K., Barnes, P.J. & Rogers D.F. (1990) Capsaicin and sensory neuropeptide stimulation of goblet cell secretion in guinea pig trachea. Journal of Physiology. 431: 629
Lemann, W., Spaer, C.B., Rye, D.B. & Wainer, B.H. (1985) Stabilization of TMB reaction product for electron microscopic retrograde and anterograde fiber tracing. Brain Research. 14: 277 - 281
Leong, S.K. & Ling, E.A. (1991) Vagus nerve and spinal cord projecting neurons demonstrated by horseradish peroxidase and different fluorescent dyes. International Journal of Neuroscience. 57: 61-72
Leong, S.K. & Tan, C.K. (1987) Central projection of rat sciatic nerve fibers as revealed by Ricinus communis agglutinin and horseradish peroxidaes tracers. Journal of Anatomy. 154: 15-26
Ling, E.A. ＆ Leong, S.K. (1987) Effects of intraneural injection of Ricinus communis agglutinin-60 into the rat vagus nerve. Journal of Neurocytology. 16: 373-387
Ling, E.A. ＆ Leong, S.K. (1988) Infiltration of carbon-labeled monocytes into the dorsal motor nucleus following an intraneural injection of Ricinus communis agglutinin-60 into the vagus nerve in rats. Journal of Anatomy. 159: 207-218
Ling, E.A., Shieh, J.Y., Wen, C.Y., Chan, Y.G. ＆ Wong, W.C. (1990) Degenerative changes of neurons in the superior cervical ganglion following an injection of Ricinus communis agglutinin-60 into the vagus nerve in hamsters. Journal of Neurocytology. 19: 1-9
Ling, E.A., Wen, C.Y., Shieh, J.Y., Yick, T.Y. ＆ Leong, S.K. (1989) Neuroglial response to neuron injury. A study using intraneural injection of Ricinus communis agglutinin-60. Journal of Anatomy. 164: 201-213
Ling, E.A., Wen, C.Y., Shieh, J.Y., Yick, T.Y. & Wong W.C. (1991) Ultrastructural changes of the nodose ganglion cells following an intraneural injection of Ricinus communis agglutinin-60 into the vagus nerve in hamsters. Journal of Anatomy. 179: 23-32
Ling, E.A. & Wong, W.C. (1988) An electron microscopic study of the nodose (inferior vagal) ganglion cells in the monky. Journal of Neurocytology. 17: 845-857
Lo, Y.K. & Huang, H.T. (1997) A novel procedure for mediastinal vagotomy inhibits neurogenic inflammation in rat bronchial tree. Journal of the Autonomic Nervous System. 67: 79-88
Lucier, G.E., Egizii, R. & Dostrovsky, J.O. (1986) Projection of the internal branch of the superior laryngeal nerve of cat. Brain Research Bulletin. 16: 713-721
Lundberg, J.M. & Saria, A. (1983) Capsaicin-induced desensitization of the airway mucosa to cigarette somke , mechanical and chemical irritants. Nature. 302: 251-253
Madey, S.M., Cole, K.J. & Brand, R.A. (1997) Sensory innervation of the cat knee articular capsule and cruciate ligament visualised using anterogradely transported wheat germ agglutinin-horseradish peroxidase. Journal of Anatomy. 190: 289-297
Massari, V.J., Johnson, T.A., Gillis, R.A. & Gatti, P.J. (1996) What are the roles of substance P and neurokinin-1 receptors in the control of negative chronotropic or negative dromotropic vagal motoneurons? A physiological and ultrastructural analysis. Brain Research. 715: 197-207
Mathsion R. & Davison J.S. (1993) Attenuated plasma extravasation to sensory neuropeptides in diabetic rats. Agents and Actions. 38: 54-59
Matsumoto, A. (1998) Synaptic changes in the perineal motoneurons of aged male rats. The Journal of Comparative Neurology. 400: 103-109
McDonald, D.M. (1988a) Neurogenic inflammation in the rat. Trachea. I : Changes in venules leucocytes and epithelium cell. Journal of Neurocytology. 17: 583-628
McDonald, D.M. (1988b) Respiratory tract infections increase susceptibility to neurogenic inflammation in the rat trachea. American Review of Respiratory Disease. 137: 1432-1440
Munoz-Martinez, E.J., Cuellar-Pedroza, L.H., Rubio-Franchini, C., Jauregui-Rincon, J. & Joseph-Nathan, P. (1994) Depression of fast axonal transport in axons demyelinated by intraneural injection of a neurotoxin from K. humboldtiana. Neurochemical Research. 19: 1341-1348
Navaratnam, V., Jacques, T.S. & Skepper, J.N. (1998) Ultrastructural and cytochemical study of neurons in the rat dorsal motor nucleus of vagus after axon crush. Microscopy Research and Technique. 42: 334-344
Nohr, D., Buob, A., Gartner, K. & Weihe, E. (1996) Changes in pulmonary calcitonin gene-related peptide and protein gene product 9.5 innervation in rats infected with Mycoplasma pulmonis. Cell and Tissue Research. 283: 215-219
Oeltmann, T.N. & Wiley, R.G. (1986) Wheat germ agglutinin-ricin A-chain conjugate is neuronotoxic after vagal injection. Brain Research. 377: 221-228
Pangalos, M.N., Francis, P.T., Pearson, R.C., Middlemiss, D.N. & Bowen, D.M. (1991) Destruction of a sub-population of cortical neurons by suicide transport of volkensin, a lectin from adenia volkensii. Journal of Neuroscience Methods. 40: 17-29
Patrickson, J.W., Smith, T.E. & Zhou, S.S. (1991) Afferent projection of the superior and recurrent laryngeal nerves. Brain Research. 539: 169-174
Poli, M.A., Rivera, V.R., Pitt, M.L & Vogel, P. (1996) Aerosolized specific antibody protects mice from lung injury associated with aerosolized ricin exposure. Toxicon. 34: 1037-1044
Puigdellivol, S.A., Prats, G. A., Ruano, G. D. & Molander, C. (2000) Fast blue and diamidino yellow as retrograde tracers in peripheral nerve: efficacy of combined nerve injection and capsule application to transected nerve in the adult rat. Journal of Neuroscience Methods. 95: 103-110
Rivero-Melian, C. & Arvidsson, J. (1989) Dose and time dependent selective and nonselective effects of ricin (RCA-120) on rat primary sensory neurons. Brain Research. 496: 131-140
Saleh, T.M. & Connell, B.J. (1998) The parabrachial nucleus mediates the decreased cardiac baroreflex sensitivity observed following short- term visceral afferent activation. Neuroscience. 87: 135-146
Soler-Rodriguez, A.M., Ghetie, M.A., Oppenheimer-Marks, N., Uhr, J.W. & Vitetta, E.S. (1993) Ricin A-chain and ricin A-chain immunotoxins rapidly damage human endothelial cells: Implications for vascular leak syndrome. Experimental Cell Research. 206: 227-234
Spurr, A.R. (1969) A low-viscosity epoxy resin embedding medium for electron microscopy. Journal of Ultrastructure Research. 26: 31-43
Tang, H.Z., Tsai, A. & Hammerschlag, R. (1994) Selective inhibition of neuronal protein synthesis by retrogradely trnsported ricin. Journal of Neuroscience Methods. 55: 15-22
Tornquist, E., Liu, L., Mattsson, P. & Svensson, M. (1997) Response of glial cells and activation of complement following motoneuron degeneration induced by toxic ricin. Neuroscience Research. 28: 167-175
Toyoda, K., Hisa, Y., Uno, T. & Tadaki, N. (1993) Distribution of the afferent neurons from the canine recurrent laryngeal nerve. European Archives of Otorhinolaryngology. 249: 485-487
Von Bartheld, C.S., Cunningham, D.E. & Rubel, E.W. (1990) Neuronal tracing with DiI: Decalcification, cryosectioning, and photoconversion for light and electron microscopic analysis. The Journal of Histochemistry and Cytochemistry. 38: 725-733
Von Bohlen, H.O. & Kiernan, J.A. (1999) Diaminobenzidine induces fluorescence in nervous tissue and provides intrinsic counterstaining of sections prepared for peroxidase histochemistry. Biochemistry and Histochemistry.74: 236-43
Wiley, R.G., Blessing, W.W. & Reis, D.J. (1982) Suicide transport: destruction of neurons by retrograde transport of ricin, abrin and modeccin. Science. 216: 889-890
Wiley, R.G. & Oeltmann, TN. (1986) Anatomically selective peripheral nerve ablation using intraneural ricin injection. Journal of Neuroscience Methods. 17: 43-53
Wilhelmsen, C.L. & Pitt, M.L.M. (1996) Lesion of acute inhaled ricin intoxication in rhesus monkeys. Veterinary Pathology. 33: 296-302
Xu, X.J., Dalsgaard, C.J., Maggi, C.A. & Wiesenfels-Hallin, Z. (1992) NK-1, but not NK-2, tachykinin receptors mediate plasma extravasation induced by antidromic C-fiber stimulation in rat hindpaw: demonstrated with the NK-1 antagonist CP-96345 and the NK-2 antagonist Men 10207. Neuroscience Letters. 139: 249-252
Yoshida, Y., Tanaka, Y., Mitsumasu, T., Hirano, M. & Kanaseki, T. (1986) Peripheral course and intramucosal distribution of the laryngeal sensory nerve fibers of cats. Brain Research Bulletin. 17: 95-105
Zhang, H.Q. & Lai, Y.L. (1993) Role of vagus nerves in resiniferatoxin-induced bronchoconstriction of guinea pigs. Respiratory Physiology. 94: 285-295