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http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/75282完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.author | Che Liu | en |
| dc.contributor.author | 劉澤 | zh_TW |
| dc.date.accessioned | 2021-07-01T08:12:30Z | - |
| dc.date.available | 2021-07-01T08:12:30Z | - |
| dc.date.issued | 2002 | |
| dc.identifier.citation | 李忠穎(2001) 大白鼠大腦反應型星形膠細胞增生之研究．國立台灣大學動物學研究所碩士論文
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/75282 | - |
| dc.description.abstract | 中間絲為直徑約10nm之細胞骨骼胞器，除了構成骨架之中間絲蛋白外，細胞中還存在有中間絲附屬蛋白(IFAP)。中間絲附屬蛋白為在形態及功能上與中間絲相關，但其本身不構成中間絲骨架之蛋白質，一般認為其功能為調節中間絲立體結構及中間絲和細胞中其他成分的交互作用。表現於成熟神經元中之中間絲蛋白有五種，包括第三類中間絲蛋白peripherin；第四類的三種neurofilament triplet proteins(NFPs)與α-internexin。這些中間絲蛋白在神經形態與功能上以及神經的疾病方面都有著重要的影響。 在背根神經節中，感覺神經元細胞本體可以依直徑與H&E染色分成染色較深之大神經元與染色較淺之小神經元兩類，這兩類細胞在形態、功能與蛋白質表現上都不盡相同。威覺神經元中的中間絲蛋白主要有NFPs、peripherin與α-internexin。NFP-H、NFP-M與NFP-L只表現在大神經元中，而peripherin與α-internexin則只在小神經元中發現。 本實驗室先前在BHK-21細胞株發現了一分子量為70-280kDa之中間絲附屬蛋白(IFAP-70/280kDa)，此蛋白與中樞神經系統及肌肉發育過程有關，其在體內的表現侷限於胚胎時期及出生一週內的輻射膠細胞及肌肉母細胞，此復上述細胞內此蛋白質的表現即告終止。然而在大白鼠成體中樞神經系統受傷以後，傷口周圍的部份反應型星狀膠細胞可以觀察到IFAP-70/280kDa之再表現於星狀膠細胞中。本研究發現成體大白鼠背根神經節中之部分神經元可以觀察到IFAP-70/280kDa的免疫螢光染色。經測量其直徑後發現此群神經元屬於大神經元。此蛋白質僅分佈於感覺神經元細胞本體(cell soma)以及剛自細胞本體延伸出來的軸突中，在遠端的軸突樹突內皆沒有表現。對實驗鼠施打cycloheximide後發現此蛋白質之表現並非受傷後之再表現。分別以NFP-L、-H、peripherin與IFAP-70/280kDa進行雙重免疫螢光染色之結果顯示IFAP-positive神經元中亦含有NFP-L及NFP-H，且以IFAP-70/280kDa與peripherin之雙重免疫螢光染色可將感覺神經元區分為三個亞族群。Immunoblotting analysis結果顯示在背根神經節IF-enriched preparation中可偵測到多個70kDa到240kDa之polypeptide,顯示背根神經節中確實含有IFAP-70/280kDa。本研究結果證實IFAP-70/280kDa為第一個發現於神經元內之中間絲附屬蛋白，同時這也是第一次在成體中觀察到IFAP-70/280kDa之表現，此蛋白於大神經元中之表現可能顯示其與神經元大小以及細胞骨骼結構之形成有關，極需進一步之研究。 | zh_TW |
| dc.description.provenance | Made available in DSpace on 2021-07-01T08:12:30Z (GMT). No. of bitstreams: 0 Previous issue date: 2002 | en |
| dc.description.tableofcontents | 謝辭……………………………………………………………………………………………………………………i 摘要……………………………………………………………………………………………………………………ii 目次……………………………………………………………………………………………………………………iv 緒論 一、中間絲……………………………………………………………………………………………………………1 二、中間絲附屬蛋白與cytoskeletal crosslinker proteins …………………………………………………3 三、神經元中之中間絲蛋白…………………………………………………………………………………………5 四、Neurofilament triplet proteins……………………………………………………………………………5 五、Peripherin………………………………………………………………………………………………………7 六、α-internexin …………………………………………………………………………………………………7 七、背根神經節………………………………………………………………………………………………………8 八、中間絲在感覺神經元中之表現…………………………………………………………………………………9 九、IFAP-70/280kDa…………………………………………………………………………………………………10 十、實驗目的…………………………………………………………………………………………………………11 實驗材料 一、實驗動物…………………………………………………………………………………………………………13 二、初級抗體…………………………………………………………………………………………………………13 三、特殊藥品…………………………………………………………………………………………………………13 實驗方法 一、石蠟包埋切片……………………………………………………………………………………………………14 二、冷凍組織切片……………………………………………………………………………………………………15 三、單一與雙重免疫螢光染色………………………………………………………………………………………15 四、BHK-21細胞株之培養……………………………………………………………………………………………16 五、IF-enriched preparation ……………………………………………………………………………………16 六、蛋白質膠體電泳…………………………………………………………………………………………………17 七、Immunoblotting Analysis ……………………………………………………………………………………18 八、Measurement of Neuron Diameters …………………………………………………………………………18 實驗結果 一、大白鼠背根神經節感覺神經元之形態觀察……………………………………………………………………20 二、IFAP-70/280kDa於大白鼠背根神經節中之表現情形…………………………………………………………21 三、IFAP-70/280kDa於施打cycloheximide大白鼠背根神經節中之表現 ………………………………………22 四、IFAP-701280kDa與NFP-L之雙重免疫螢光染色 ………………………………………………………………23 五、IFAP-70/80kDa與NFP-H之雙重免疫螢光染色…………………………………………………………………24 六、IFAP-70/280kDa與Peripherin之雙重免疫螢光染色…………………………………………………………25 七、Immunoblotting analysis ……………………………………………………………………………………25 八、感覺神經中無Vimentin與Nestin………………………………………………………………………………26 討論 一、感覺神經元直徑之測量…………………………………………………………………………………………27 二、中間絲附屬蛋白IFAP-70/280kDa在大白鼠背根神經節中之表現……………………………………………28 三、中間絲蛋白於感覺神經元中之功能……………………………………………………………………………31 四、IFAP-70/280kD表現於感覺神經元中之可能意義 ……………………………………………………………32 五、結語：神經元內發現的第一個中間絲附屬蛋白………………………………………………………………34 參考文獻………………………………………………………………………………………………………………36 圖表……………………………………………………………………………………………………………………45 | |
| dc.language.iso | zh-TW | |
| dc.title | IFAP-70/280kD於大白鼠腰椎背根神經節中之表現 | zh_TW |
| dc.title | Expression of IFAP-70/280kD in Large Neurons of Rat Lumbar Dorsal Root Ganglia | en |
| dc.date.schoolyear | 90-2 | |
| dc.description.degree | 碩士 | |
| dc.relation.page | 73 | |
| dc.rights.note | 未授權 | |
| dc.contributor.author-dept | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 動物學研究所 | zh_TW |
| 顯示於系所單位： | 動物學研究所 | |
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