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研究生:劉峰銓
研究生(外文):Feng-Chuan Liu
論文名稱:白點症病毒感染草蝦之胃與類淋巴器官間細胞凋亡之比較分析
論文名稱(外文):Comparative analysis of apoptosis between stomach and lymphoid organ in white spot syndrome virus infected Penaeus monodon
指導教授:郭光雄羅竹芳羅竹芳引用關係
指導教授(外文):Guang-Hsiung KouChu-Fang Lo
學位類別:碩士
校院名稱:國立臺灣大學
系所名稱:動物學研究研究所
學門:生命科學學門
學類:生物學類
論文種類:學術論文
論文出版年:2004
畢業學年度:92
語文別:中文
論文頁數:62
中文關鍵詞:細胞凋亡白點症病毒
外文關鍵詞:apoptosiswhite spot syndrome virus
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細胞凋亡是多細胞生物用來維持細胞恆定性和消除對自體有潛在危險性細胞的保護反應。宿主可啟動細胞凋亡機制讓受病毒感染的細胞死亡以限制病毒複製。另方面,病毒可調控細胞凋亡路徑延緩宿主細胞死亡以完成病毒複製或建立持續感染。本實驗將白點症病毒(White spot syndrome virus, WSSV)帶原草蝦(Penaeus monodon)及無WSSV帶原草蝦經肌肉注射WSSV或PBS為對照組。於不同時期取三個獨立樣本的胃和類淋巴組織,進行WSSV原位雜合反應(in situ hybridization)與細胞凋亡檢測(TUNEL assay),以陽性反應細胞的百分比數進行定量分析組織內WSSV DNA複製與細胞凋亡發生程度。結果顯示草蝦於人工感染WSSV 72小時後, 於類淋巴器官內的TUNEL陽性反應細胞較胃高;相對地,類淋巴器官內的WSSV原位雜合陽性反應細胞較胃低。此結果為WSSV複製與細胞凋亡相互關係之日後研究奠定了基礎。
Apoptosis is a genetically controlled multicellular organisms process to maintain homeostasis and remove the potentially dangerous cells. The host can exterminate the infected cells base on activating the apoptosis system and thus suppress virus reproduction. On the other hand, viruses could replicate themselves or cause persistent infection by regulating the apoptotic pathway to delay the death of host cell. The previous investigations indicate that apoptosis occurs in white spot syndrome virus (WSSV) infected Penaeus monodon. In this study, WSSV-carriers and WSSV-free shrimp were intramuscularly injected with WSSV or PBS vehicle as controls. Moreover, three independent specimens of stomachs and lymphoid organs (LO) were then collected in different time points to examine apoptosis incurrence and levels of WSSV DNA replication through TUNEL assay and WSSV in situ hybridization (ISH) respectively. The findings reveal that the TUNEL positive cells were higher in the LO than in the stomach at 72 hours after WSSV infection. In contrast, the ISH positive cells were lower in the LO than in the stomach. The results facilitate further studies relationship between apoptosis and WSSV replication.
第壹章 前 言

蝦白點症病毒…………………………………………… 1
蝦白點症病毒檢測……………………………………… 3
細胞凋亡………………………………………………… 4
細胞凋亡偵測…………………………………………… 6
研究目的………………………………………………… 7

笫貳章 材 料 與 方 法

一、草蝦來源與檢測…………………………………… 9
二、人工感染帶原草蝦和無帶原草蝦………………… 9
三、採集樣本包埋與組織切片………………………… 10
四、白點症病毒原位雜合之核酸探針製備…………… 10
五、白點症病毒原位雜合探針最佳反應濃度………… 11
六、白點症病毒原位雜合反應分析…………………… 12
七、細胞凋亡反應分析………………………………… 12
八、白點症病毒與細胞凋亡關聯性之統計分析……… 13

第參章 結 果

一、白點症病毒原位雜合反應分析…………………… 14
二、細胞凋亡反應訊號分析…………………………… 15
三、白點症病毒與細胞凋亡關聯性之統計分析……… 16

第肆章 討 論

討論……………………………………………………… 19

參考文獻………………………………………………… 25
表圖……………………………………………………… 38
附錄……………………………………………………… 49
參與發表論文…………………………………………… 62
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