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研究生:陳先鋒
論文名稱:重組腺相關病毒作為新的神經追蹤物探討兔頸上神經節傳入神經元
論文名稱(外文):Recombinant Adeno-Associated Virus as a Novel Tracer for Studying Afferent Neurons of the Superior Cervical Ganglion in Rabbits
指導教授:王順德劉江川
學位類別:碩士
校院名稱:國防醫學院
系所名稱:生物及解剖學研究所
學門:生命科學學門
學類:生物訊息學類
論文種類:學術論文
論文出版年:1997
畢業學年度:85
語文別:中文
論文頁數:48
中文關鍵詞:腺相關病毒過氧化氫酵素頸上神經節大白兔
外文關鍵詞:adeno-associated virusHRPsuperior cervical ganglionrabbit
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近年來,病毒經常被使用來作為神經徑路追蹤劑,乃因病毒會感染神經元並且會在神經元內複製。病毒雖然經由分子生物技術的修飾以降低毒性,但卻仍然含有殘餘的毒性與致病的能力,所以這些病毒的使用因而受到限制,故發展一安全又穩定的病毒做為神經追蹤物是相當重要的。重組腺相關病毒,屬於小病毒,其為單股去氧核醣核酸病毒,具有做為神經元的病毒性神經追蹤劑有利的特性如下:非致病性、穩定、對於不再分裂的神經元細胞能夠感染並將其帶有的基因片段送入細胞內。本實驗目的是研究重組腺相關病毒是否能做為一嶄新的病毒性神經追蹤劑,並藉以探討大白兔額上神經節傳入神經元來源。將重組腺相關病毒及過氧化氫酵素 (horseradish peroxidase; HRP) 注入紐西蘭大白兔單側頸上神經節,實驗動物存活三天之後,將實驗動物犧牲,取出雙側的頸上神經節、星狀神經節、三叉神經節、nodose神經節以及從第一頸脊髓至第一腰脊髓段之脊髓與雙側背根神經節並以冷凍切片機將標本切片,然後進行HRP及β-GaI組織化學反應。結果顯示:(1) 被 HRP 標示的神經元位置在從脊髓第八頸脊能到第七胸脊髓段中的中間外側核(intermediolateral nucleus: IML)、同側星狀神經節 (ipsilateral stellate ganglia),(2) 有重組腺相關病毒陽性反應 (AAV positive) 的神經元位置從第一頸脊髓到第一腰脊髓段之雙側背根神經節、坐側頸上神經節以及雙側三叉神經節。從本實驗所獲得的結果可推測,重組腺相關病毒能夠感染感覺性神經元,但似乎是無法感染交感神經系統的節前神經元。
Recently, viruses are wildly used for experimental neural tract tracing because they can infect and replicate within neurons and are transported within the axons. However, issues of continued virus gene expression, pathogenicity and reversion to wild type still remain. The recombinant adeno-associated virus (AAV), a virus with a single strain DNA, is considerable a safe neural tracer because of its special properties of (1) the specific integration, non-pathogenecity of wild-type and non-replication without helper virus and (2) AAV could infect the non-mitotic neurons. The purposes of this study are : (1) to evaluate the efficacy of AAV to be as a novel neural tracer; (2) to study the origins of the afferent fiber of the superior cervical ganglion (SCG) of the rabbits. AAV and /or HRP were injected into the unilateral SCG of the rabbits. Three days after the tracer injected, rabbits were sacrificed by perfusion with Ringer's solution and fixative. The bilateral SCG, stellate ganglion, nodose ganglion, trigeminal ganglion, dorsal root ganglia and the spinal cord from C1 to L1 were removed and sectioned, then processed for HRP and β-Gal histochemical reactions. Results show that: (1) the HRP labeling neurons were located in the ipsilateral intermediolateral nucleus (IML) of the spinal segments from C8 to T7 and the ipsilateral stellate ganglion; (2) the AAV positive neurons were found in bilateral SCG, trigeminal ganglion and dorsal root ganglia from C1 to L5 but not in the IML and stellate ganglion. Results suggest that AAV appeared to be able to label the sensory neurons rather than the sympathetic preganglionic neurons.
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